Official Monographs PDF

Official Monographs
water, 1-butanol and acetic acid (100) (5:4:1) to a distance of

Acebutolol Hydrochloride about 10 cm, and air-dry the plate. Examine under ultravio-
let light (main wavelength: 365 nm): the spots other than the
アセブトロール塩酸塩 principal spot from the sample solution are not more intense
than the spot from the standard solution.
Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C,
3 hours).
Residue on ignition <2.44> Not more than 0.2z (1 g).
Assay Weigh accurately about 0.25 g of Acebutolol Hydro-
C18H28N2O4.HCl: 372.89 chloride, previously dried, dissolve in 20 mL of acetic acid
N-{3-Acetyl-4-[(2RS )-2-hydroxy- (100), add 80 mL of acetic anhydride, and titrate <2.50> with
3-(1-methylethyl)aminopropyloxy]phenyl}butanamide 0.1 mol/L perchloric acid VS (potentiometric titration). Per-
monohydrochloride form a blank determination, and make any necessary correc-
[34381-68-5] tion.
Each mL of 0.1 mol/L perchloric acid VS
Acebutolol Hydrochloride, when dried, contains ＝ 37.29 mg of C18H28N2O4.HCl
not less than 98.0z and not more than 102.0z of
acebutolol hydrochloride (C18H28N2O4.HCl). Containers and storage Containers—Well-closed contain-
ers.
Description Acebutolol Hydrochloride occurs as white to
pale yellowish white, crystals or crystalline powder.
It is freely soluble in water, in methanol, in ethanol (95)
and in acetic acid (100), and practically insoluble in diethyl Aceglutamide Aluminum
ether.
アセグルタミドアルミニウム
A solution of Acebutolol Hydrochloride (1 in 20) shows
no optical rotation.
Identification (1) Determine the absorption spectrum of a
solution of Acebutolol Hydrochloride in 0.01 mol/L hydro-
chloric acid TS (1 in 100,000) as directed under Ultraviolet-
visible Spectrophotometry <2.24>, and compare the spectrum
with the Reference Spectrum: both spectra exhibit similar in-
tensities of absorption at the same wavelengths.
C35H59Al3N10O24: 1084.84
(2) Determine the infrared absorption spectrum of
Pentakis[(2S )-2-acetylamino-4-
Acebutolol Hydrochloride, previously dried, as directed in
carbamoylbutanoato]tetrahydroxotrialuminium
the potassium bromide disk method under Infrared Spectro-
[12607-92-0]
photometry <2.25>, and compare the spectrum with the Ref-
erence Spectrum: both spectra exhibit similar intensities of
Aceglutamide Aluminum contains not less than
absorption at the same wave numbers.
85.4z and not more than 87.6z of aceglutamide
(3) A solution of Acebutolol Hydrochloride (1 in 100)
(C7H12N2O4: 188.18), and not less than 7.0z and not
responds to the Qualitative Tests <1.09> for chloride.
more than 8.0z of aluminum (Al: 26.98), calculated
Melting point <2.60> 141 – 1459C on the dried basis.
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Description Aceglutamide Aluminum occurs as a white
Acebutolol Hydrochloride according to Method 2, and powder, having astringent bitter taste.
perform the test. Prepare the control solution with 1.0 mL of It is freely soluble in water, and practically insoluble in
Standard Lead Solution (not more than 10 ppm). ethanol (99.5).
(2) Arsenic <1.11>—Prepare the test solution with 1.0 g It dissolves in dilute hydrochloric acid.
of Acebutolol Hydrochloride according to Method 3, and It is hygroscopic.
perform the test (not more than 2 ppm).
Identification (1) Dissolve 0.03 g each of Aceglutamide
(3) Related substances—Dissolve 40 mg of Acebutolol
Aluminum and Aceglutamide RS in 5 mL of water, and use
Hydrochloride in 2 mL of methanol, and use this solution as
these solutions as the sample solution and standard solution.
the sample solution. Pipet 1 mL of the sample solution, add
Perform the test with these solutions as directed under Thin-
methanol to make exactly 25 mL, and pipet 1 mL of this so-
layer Chromatography <2.03>. Spot 5 mL each of the sample
lution, add methanol to make exactly 20 mL, and use this so-
solution and standard solution on a plate of cellulose for
lution as the standard solution. Perform the test with these
thin-layer chromatography. Develop the plate with a mixture
solutions as directed under Thin-layer Chromatography
of 1-propanol, water and acetic acid (100) (16:8:1) to a dis-
<2.03>. Spot 5 mL each of the sample solution and standard
tance of about 10 cm, and air-dry the plate. Spray evenly a
solution on a plate of silica gel for thin-layer chromatogra-
solution of bromocresol green in ethanol (95) (1 in 1000),
phy. Develop the plate with the upper layer of a mixture of
then spray evenly diluted ammonia solution (28) (1 in 100):
The JP Drugs are to be tested according to the provisions given in the pertinent monographs, General Notices, General Rules for Crude Drugs, 359
General Rules for Preparations, and General Tests for their conformity to the Japanese Pharmacopoeia. (See the General Notices 5.)
360 Aceglutamide Aluminum / Official Monographs JP XVII
the spots from the sample solution and the standard solution Loss on drying <2.41> Not more than 5.0z (1 g, 1309C,
show a light yellow and have the same R f value. 5 hours).
(2) A solution of Aceglutamide Aluminum in dilute hy-
Assay (1) Aceglutamide—Weigh accurately about 50 mg
drochloric acid (1 in 20) responds to the Qualitative Tests
of Aceglutamide Aluminum, dissolve in a suitable amount of
<1.09> for aluminum salt.
the mobile phase, add exactly 10 mL of the internal standard
Optical rotation <2.49> [a]20
D : －5.5 – －7.59(2 g calculated solution and the mobile phase to make 50 mL, and use this
on the dried basis, water, 50 mL, 100 mm). solution as the sample solution. Separately, weigh accurately
about 45 mg of Aceglutamide RS, dissolve in a suitable
Purity (1) Heavy metals <1.07>—Put 1.0 g of Acegluta-
amount of the mobile phase, add exactly 10 mL of the inter-
mide Aluminum in a porcelain crucible, cover the crucible
nal standard solution and the mobile phase to make 50 mL,
loosely, and heat gently to carbonize. After cooling, add 2
and use this solution as the standard solution. Perform the
mL of nitric acid and 1 mL of sulfuric acid, heat gently until
test with 10 mL each of the sample solution and standard so-
the white fumes no more evolve, and heat to incinerate at
lution as directed under Liquid Chromatography <2.01> ac-
500 to 6009 C. If the incineration is not accomplished, add 2
cording to the following conditions, and calculate the ratios,
mL of nitric acid and 1 mL of sulfuric acid, heat in the same
QT and QS, of the peak area of aceglutamide to that of the
manner as above, then ignite at 500 to 6009C to incinerate.
internal standard.
After cooling, add 2 mL of hydrochloric acid, proceed with
this solution according to Method 2, and perform the test. Amount (mg) of aceglutamide (C7H12N2O4)
Prepare the control solution as follows: proceed in the same ＝ MS × QT/QS
manner as the preparation of the test solution with the same
MS: Amount (mg) of Aceglutamide RS taken
amount of the reagents, and add 2.0 mL of Standard Lead
Solution and water to make 50 mL (not more than 20 ppm). Internal standard solution—A solution of thymine in metha-
(2) Arsenic <1.11>—Prepare the test solution with 1.0 g nol (1 in 4000).
of Aceglutamide Aluminum according to Method 1, and per- Operating conditions—
form the test (not more than 2 ppm). Detector: An ultraviolet absorption photometer (wave-
(3) Related substances—Dissolve 0.10 g of Aceglutamide length: 210 nm).
Aluminum in the mobile phase to make exactly 100 mL, and Column: A stainless steel column 4.6 mm in inside diame-
use this solution as the sample solution. Pipet 1 mL of the ter and 25 cm in length, packed with octadecylsilanized silica
sample solution, add the mobile phase to make exactly 100 gel for liquid chromatography (5 mm in particle diameter).
mL, and use this solution as the standard solution (1). Sepa- Column temperature: A constant temperature of about
rately, dissolve 10 mg of 2-acetamidoglutarimide in the mo- 259C.
bile phase to make exactly 100 mL. Pipet 3 mL of this solu- Mobile phase: A mixture of diluted perchloric acid (1 in
tion, add the mobile phase to make exactly 100 mL, and use 1000) and methanol (99:1).
this solution as the standard solution (2). Perform the test Flow rate: Adjust so that the retention time of acegluta-
with exactly 20 mL each of the sample solution and standard mide is about 5 minutes.
solutions (1) and (2) as directed under Liquid Chromatogra- System suitability—
phy <2.01> according to the following conditions, and deter- System performance: When the procedure is run with 10
mine each peak area by the automatic integration method: mL of the standard solution under the above operating con-
the peak area of 2-acetamidoglutarimide from the sample ditions, aceglutamide and the internal standard are eluted in
solution is not larger than that from the standard solution this order with the resolution between these peaks being not
(2), the peak areas other than aceglutamide and 2-acetamido- less than 11.
glutarimide from the sample solution are not larger than System repeatability: When the test is repeated 6 times
3/10 times the peak area of aceglutamide from the standard with 10 mL of the standard solution under the above operat-
solution (1), and the total of the peak areas other than ace- ing conditions, the relative standard deviation of the ratios
glutamide and 2-acetamidoglutarimide from the sample solu- of the peak area of aceglutamide to that of the internal
tion is not larger than the peak area of aceglutamide from standard is not more than 1.0z.
the standard solution (1). (2) Aluminum—Weigh accurately about 3.0 g of Aceglu-
Operating conditions— tamide Aluminum, add 20 mL of dilute hydrochloric acid,
Detector, column, column temperature, mobile phase, and and heat on a water bath for 60 minutes. After cooling, add
flow rate: Proceed as directed in the operating conditions in water to make exactly 200 mL. Pipet 20 mL of this solution,
the Assay. add exactly 25 mL of 0.05 mol/L disodium dihydrogen
Time span of measurement: About 3 times as long as the ethylenediamine tetraacetate VS and 20 mL of acetic acid-
retention time of aceglutamide. ammonium acetate buffer solution (pH 4.8) and boil for 5
System suitability— minutes. After cooling, add 50 mL of ethanol (95), and
System performance: Proceed as directed in the system titrate <2.50> with 0.05 mol/L zinc acetate VS until the color
suitability in the Assay (1). of the solution changes from light dark green to light red (in-
Test for required detectability: To exactly 5 mL of the dicator: 2 mL of dithizone TS). Perform a blank determina-
standard solution (1) add the mobile phase to make exactly tion in the same manner.
50 mL. Confirm that the peak area of aceglutamide obtained
Each mL of 0.05 mol/L disodium dihydrogen
from 20 mL of this solution is equivalent to 7 to 13z of that
ethylenediamine tetraacetate VS ＝ 1.349 mg of Al
of aceglutamide obtained from 20 mL of the standard solu-
tion. Containers and storage Containers—Tight containers.
System repeatability: When the test is repeated 6 times
with 20 mL of the standard solution (1) under the above
operating conditions, the relative standard deviation of the
peak area of aceglutamide is not more than 2.0z.
The JP Drugs are to be tested according to the provisions given in the pertinent monographs, General Notices, General Rules for Crude Drugs,
JP XVII Official Monographs / Acemetacin Capsules 361

Acemetacin Assay Weigh accurately about 0.35 g of Acemetacin, previ-
ously dried, dissolve in 20 mL of acetone, add 10 mL
アセメタシン
of water, and then titrate <2.50> with 0.1 mol/L sodium hy-
droxide VS (potentiometric titration). Perform a blank de-
termination in the same method, and make any necessary
correction.
Each mL of 0.1 mol/L sodium hydroxide VS
＝ 41.58 mg of C21H18ClNO6
Containers and storage Containers—Tight containers.
C21H18ClNO6: 415.82
2-{2-[1-(4-Chlorobenzoyl)-5-methoxy-2-methyl-1H-
indol-3-yl]acetyloxy}acetic acid Acemetacin Capsules
[53164-05-9]
アセメタシンカプセル
Acemetacin, when dried, contains not less than
99.0z and not more than 101.0z of acemetacin Acemetacin Capsules contain not less than 93.0z
(C21H18ClNO6). and not more than 107.0z of the labeled amount of
acemetacin (C21H18ClNO6: 415.82).
Description Acemetacin occurs as a light yellow crystalline
powder. Method of preparation Prepare as directed under Cap-
It is soluble in acetone, sparingly soluble in methanol, sules, with Acemetacin.
slightly soluble in ethanol (99.5), and practically insoluble in
Identification To an amount of powdered contents of
water.
Acemetacin Capsules, equivalent to 0.1 g of Acemetacin,
Identification (1) To 1 mg of Acemetacin add 1 mL of add 100 mL of methanol, shake well, and filter. Take 10 mL
concentrated chromotropic acid TS, and heat in a water bath of the filtrate, and distil the methanol under reduced pres-
for 5 minutes: a red-purple color develops. sure. To the residue add 1 mL of methanol, shake well,
(2) Determine the absorption spectrum of a solution of centrifuge, and use the supernatant liquid as the sample
Acemetacin in methanol (1 in 50,000) as directed under Ul- solution. Separately, dissolve 10 mg of acemetacin in 1 mL
traviolet-visible Spectrophotometry <2.24>, and compare the of methanol, and use this solution as the standard solution.
spectrum with the Reference Spectrum: both spectra exhibit Perform the test with these solutions as directed under Thin-
similar intensities of absorption at the same wavelengths. layer Chromatography <2.03>. Spot 2 mL each of the sample
(3) Determine the infrared absorption spectrum of solution and standard solution on a plate of silica gel with
Acemetacin as directed in the potassium bromide disk fluorescent indicator for thin-layer chromatography. De-
method under Infrared Spectrometry <2.25>, and compare velop the plate with a mixture of hexane, 4-methyl-2-penta-
the spectrum with the Reference Spectrum: both spectra none and acetic acid (100) (3:2:1) to a distance of about 10
exhibit similar intensities of absorption at the same wave cm, and air-dry the plate. Examine under ultraviolet light
numbers. (main wavelength: 254 nm): the spots obtained from the
(4) Perform the test with Acemetacin as directed under sample solution and standard solution show the same R f
Flame Coloration Test <1.04> (2): a green color appears. value.
Melting point <2.60> 151 – 1549C Uniformity of dosage units <6.02> Perform the test accord-
ing to the following method: it meets the requirement of the
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
Content uniformity test.
Acemetacin according to Method 4, and perform the test.
Take out the contents of 1 capsule of Acemetacin Cap-
Prepare the control solution with 2.0 mL of Standard Lead
sules, add 40 mL of methanol, shake well, and add methanol
Solution (not more than 20 ppm).
to make exactly V mL so that each mL contains about 0.6
(2) Related substances—Dissolve 0.40 g of Acemetacin
mg of acemetacin (C21H18ClNO6). Filter this solution, dis-
in 10 mL of acetone, and use this solution as the sample so-
card the first 10 mL of the filtrate, pipet 5 mL of the subse-
lution. Pipet 1 mL of the sample solution, and add acetone
quent filtrate, add exactly 2 mL of the internal standard so-
to make exactly 50 mL. Pipet 1 mL of this solution, add ace-
lution, add methanol to make 50 mL, and use this solution
tone to make exactly 10 mL, and use this solution as the
as the sample solution. Proceed as directed in the Assay.
standard solution. Perform the test with these solutions as
directed under Thin Layer Chromatography <2.03>. Spot Amount (mg) of acemetacin (C21H18ClNO6)
5 mL each of the sample solution and standard solution on a ＝ MS × QT/QS × V/50
plate of silica gel with fluorescent indicator for thin layer
MS: Amount (mg) of acemetacin for assay taken
chromatography. Develop the plate with a mixture of
hexane, 4-methyl-2-pentanone and acetic acid (100) (3:2:1) Internal standard solution—A solution of hexyl parahy-
to a distance of about 10 cm, and air-dry the plate. Examine droxybenzoate in methanol (1 in 1000).
under ultraviolet light (main wavelength: 254 nm): not more
Dissolution <6.10> When the test is performed at 50 revolu-
than 2 spots other than the principal spot appear from the
tions per minute according to the Paddle method using the
sample solution, and these spots are not more intense than
sinker, using 900 mL of 2nd fluid for dissolution test as the
the spot obtained from the standard solution.
dissolution medium, the dissolution rate in 30 minutes of
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, Acemetacin Capsules is not less than 70z.
2 hours). Start the test with 1 capsule of Acemetacin Capsules, with-
362 Acemetacin Tablets / Official Monographs JP XVII
draw not less than 20 mL of the medium at the specified 20 mL of this solution under the above operating conditions,
minute after starting the test, and filter through a membrane acemetacin, indometacin and the internal standard are eluted
filter with a pore size not exceeding 0.45 mm. Discard the in this order with the resolutions between the peaks of
first 10 mL of the filtrate, pipet V mL of the subsequent acemetacin and indometacin and between the peaks of
filtrate, add the dissolution medium to make exactly V? mL indometacin and the internal standard being not less than 3,
so that each mL contains about 33 mg of acemetacin respectively.
(C21H18ClNO6), and use this solution as the sample solution. System repeatability: When the test is repeated 6 times
Separately, weigh accurately about 17 mg of acemetacin for with 20 mL of the standard solution under the above operat-
assay, previously dried at 1059C for 2 hours, dissolve in the ing conditions, the relative standard deviation of the ratio of
dissolution medium to make exactly 100 mL. Pipet 4 mL of the peak area of acemetacin to that of the internal standard
this solution, add the dissolution medium to make exactly 20 is not more than 1.0z.
mL, and use this solution as the standard solution. Deter-
mine the absorbances, AT and AS, of the sample solution
and standard solution at 319 nm as directed under Ultravio-
let-visible Spectrophotometry <2.24>.
Acemetacin Tablets
Dissolution rate (z) with respect to the labeled amount
of acemetacin (C21H18ClNO6) アセメタシン錠
＝ MS × AT/AS × V?/V × 1/C × 180
MS: Amount (mg) of acemetacin for assay taken Acemetacin Tablets contain not less than 93.0z and
C: Labeled amount (mg) of acemetacin (C21H18ClNO6) in not more than 107.0z of the labeled amount of
1 capsule acemetacin (C21H18ClNO6: 415.82).
Assay Take out the contents of not less than 20 Acemeta- Method of preparation Prepare as directed under Tablets,
cin Capsules, weigh accurately the mass of the contents, and with Acemetacin.
powder. Weigh accurately a portion of the powder, equiva-
Identification To a quantity of powdered Acemetacin
lent to about 30 mg of acemetacin (C21H18ClNO6), add 40
Tablets, equivalent to 0.1 g of Acemetacin, add 100 mL of
mL of methanol, shake well, and add methanol to make
methanol, shake well, and filter. Take 10 mL of the filtrate,
exactly 50 mL. Filter this solution, discard the first 10 mL of
and distil the methanol under reduced pressure. Dissolve the
the filtrate, pipet 5 mL of the subsequent filtrate, add exactly
residue in 1 mL of methanol, centrifuge, and use the super-
2 mL of the internal standard solution, add methanol to
natant liquid as the sample solution. Separately, dissolve 10
make 50 mL, and use this solution as the sample solution.
mg of acemetacin in 1 mL of methanol, and use this solution
Separately, weigh accurately about 30 mg of acemetacin for
as the standard solution. Perform the test with these solu-
assay, previously dried at 1059 C for 2 hours, and dissolve in
tions as directed under Thin-layer Chromatography <2.03>.
methanol to make exactly 50 mL. Pipet 5 mL of this solu-
Spot 2 mL each of the sample solution and standard solution
tion, add exactly 2 mL of the internal standard solution, add
on a plate of silica gel with fluorescent indicator for thin-
methanol to make 50 mL, and use this solution as the stand-
layer chromatography. Develop the plate with a mixture of
ard solution. Perform the test with 20 mL each of the sample
hexane, 4-methyl-2-pentanone and acetic acid (100) (3:2:1)
solution and standard solution as directed under Liquid
to a distance of about 10 cm, and air-dry the plate. Examine
Chromatography <2.01> according to the following condi-
under ultraviolet light (main wavelength: 254 nm): the spots
tions, and calculate the ratios, QT and QS, of the peak area
obtained from the sample solution and standard solution
of acemetacin to that of the internal standard.
show the same R f value.
Amount (mg) of acemetacin (C21H18ClNO6)
Uniformity of dosage units <6.02> Perform the test accord-
＝ MS × QT/QS
MS: Amount (mg) of acemetacin for assay taken Content uniformity test.
To 1 tablet of Acemetacin Tablets add 3 mL of water, and
Internal standard solution—A solution of hexyl parahy-
shake until the tablet is disintegrated. Add 15 mL of metha-
droxybenzoate in methanol (1 in 1000).
nol, shake for 20 minutes, and add methanol to make exactly
Operating conditions—
V mL so that each mL contains about 1.2 mg of acemetacin
Detector: An ultraviolet absorption photometer (wave-
(C21H18ClNO6). Centrifuge this solution, filter the superna-
length 254 nm).
tant liquid, discard the first 10 mL of the filtrate, pipet 5 mL
Column: A stainless steel column 4.6 mm in inside diame-
of the subsequent filtrate, add exactly 1 mL of the internal
ter and 25 cm in length, packed with octadecylsilanized silica
standard solution, add methanol to make 50 mL, and use
gel for liquid chromatography (5 mm in particle diameter).
this solution as the sample solution. Proceed as directed in
Column temperature: A constant temperature of about
the Assay.
409 C.
Mobile phase: To 6 g of acetic acid (100) add water to Amount (mg) of acemetacin (C21H18ClNO6)
make 1000 mL, and adjust the pH to 3.2 with a solution of ＝ MS × QT/QS × V/25
1.36 g of sodium acetate trihydrate in 100 mL of water. To
200 mL of this solution add 300 mL of acetonitrile.
Flow rate: Adjust so that the retention time of acemetacin Internal standard solution—A solution of hexyl parahy-
is about 7 minutes. droxybenzoate in methanol (1 in 250).
System suitability—
System performance: Dissolve 75 mg of acemetacin and 75
tions per minute according to the Paddle method, using
mg of indometacin in 50 mL of methanol. To 2 mL of this
900 mL of 2nd fluid for dissolution test as the dissolution
solution add 2 mL of the internal standard solution, and add
medium, the dissolution rate in 45 minutes of Acemetacin
methanol to make 50 mL. When the procedure is run with
JP XVII Official Monographs / Acetaminophen 363
Tablets is not less than 80z. mg of indometacin in 50 mL of methanol. To 4 mL of this

Start the test with 1 tablet of Acemetacin Tablets, with- solution add 1 mL of the internal standard solution, and add
draw not less than 20 mL of the medium at the specified methanol to make 50 mL. When the procedure is run with
minute after starting the test, and filter through a membrane 10 mL of this solution under the above operating conditions,
filter with a pore size not exceeding 0.45 mm. Discard the acemetacin, indometacin and the internal standard are eluted
first 10 mL of the filtrate, pipet V mL of the subsequent in this order with the resolutions between the peaks of
filtrate, add the dissolution medium to make exactly V? mL acemetacin and indometacin and between the peaks of
so that each mL contains about 33 mg of acemetacin indometacin and the internal standard being not less than 3,
(C21H18ClNO6), and use this solution as the sample solution. respectively.
Separately, weigh accurately about 17 mg of acemetacin for System repeatability: When the test is repeated 6 times
assay, previously dried at 1059C for 2 hours, dissolve in the with 10 mL of the standard solution under the above operat-
dissolution medium to make exactly 100 mL. Pipet 4 mL of ing conditions, the relative standard deviation of the ratio of
this solution, add the dissolution medium to make exactly 20 the peak area of acemetacin to that of the internal standard
mL, and use this solution as the standard solution. Deter- is not more than 1.0z.
Dissolution rate (z) with respect to the labeled amount Acetaminophen
of acemetacin (C21H18ClNO6)
＝ MS × AT/AS × V?/V × 1/C × 180 Paracetamol
アセトアミノフェン
C: Labeled amount (mg) of acemetacin (C21H18ClNO6) in
1 tablet
Assay Weigh accurately the mass of not less than 20
Acemetacin Tablets, and powder. Weigh accurately a por-
tion of the powder, equivalent to about 0.6 g of acemetacin
(C21H18ClNO6), add 120 mL of methanol, shake for 20 C8H9NO2: 151.16
minutes, and add methanol to make exactly 200 mL. Centri- N-(4-Hydroxyphenyl)acetamide
fuge this solution, filter the supernatant liquid, discard the [103-90-2]
first 10 mL of the filtrate, pipet 2 mL of the subsequent
filtrate, add exactly 1 mL of the internal standard solution, Acetaminophen, when dried, contains not less than
add methanol to make 50 mL, and use this solution as the 98.0z of acetaminophen (C8H9NO2).
sample solution. Separately, weigh accurately about 30 mg
Description Acetaminophen occurs as white, crystals or
of acemetacin for assay, previously dried at 1059C for 2
crystalline powder.
hours, and dissolve in methanol to make exactly 25 mL.
It is freely soluble in methanol and in ethanol (95), spar-
Pipet 5 mL of this solution, add exactly 1 mL of the internal
ingly soluble in water, and very slightly, soluble in diethyl
standard solution, add methanol to make 50 mL, and use
ether.
this solution as the standard solution. Perform the test with
It dissolves in sodium hydroxide TS.
10 mL each of the sample solution and standard solution as
directed under Liquid Chromatography <2.01> according to Identification Determine the infrared absorption spectra of
the following conditions, and calculate the ratios, QT and Acetaminophen, previously dried, as directed in the potas-
QS, of the peak area of acemetacin to that of the internal sium bromide disk method under Infrared Spectrophotome-
standard. try <2.25>, and compare the spectrum with the Reference
Spectrum or the spectrum of dried Acetaminophen RS: both
Amount (mg) of acemetacin (C21H18ClNO6)
spectra exhibit similar intensities of absorption at the same
＝ MS × QT/QS × 20
wave numbers.
Melting point <2.60> 169 – 1729C
Internal standard solution—A solution of hexyl parahy-
Purity (1) Chloride <1.03>—Dissolve 4.0 g of Acetamino-
phen in 100 mL of water by heating, cool with shaking in ice
water, allow to stand until ordinary temperature is attained,
add water to make 100 mL, and filter. To 25 mL of the
length 254 nm).
filtrate add 6 mL of dilute nitric acid and water to make 50
mL, and perform the test using this solution as the test solu-
tion. Prepare the control solution with 0.40 mL of 0.01
mol/L hydrochloric acid VS (not more than 0.014z).
(2) Sulfate <1.14>—To 25 mL of the filtrate obtained in
409 C.
(1) add 1 mL of dilute hydrochloric acid and water to make
Mobile phase: To 6 g of acetic acid (100) add water to
50 mL, and perform the test using this solution as the test
make 1000 mL, and adjust the pH to 3.2 with a solution of
solution. Prepare the control solution with 0.40 mL of
1.36 g of sodium acetate trihydrate in 100 mL of water. To
0.005 mol/L sulfuric acid VS (not more than 0.019z).
(3) Heavy metals <1.07>—Proceed with 2.0 g of Acet-
Flow rate: Adjust so that the retention time of acemetacin
aminophen according to Method 4, and perform the test.
is about 7 minutes.
System performance: Dissolve 75 mg of acemetacin and 75
364 Acetazolamide / Official Monographs JP XVII
(4) Arsenic <1.11>—Prepare the test solution with 1.0 g
of Acetaminophen according to Method 3, and perform the Acetazolamide
test (not more than 2 ppm).
(5) Related substances—Dissolve 50 mg of Acetamino- アセタゾラミド
phen in 1 mL of methanol, add the mobile phase to make 50
mL, and use this solution as the sample solution. Pipet 1 mL
of the sample solution, add the mobile phase to make exactly
200 mL, and use this solution as the standard solution. Per-
form the test with exactly 10 mL each of the sample solution
and standard solution as directed under Liquid Chromatog-
raphy <2.01> according to the following conditions. Deter-
C4H6N4O3S2: 222.25
mine each peak area of both solutions by the automatic
N-(5-Sulfamoyl-1,3,4-thiadiazol-2-yl)acetamide
integration method: the total area of all peaks other than
[59-66-5]
acetaminophen from the sample solution is not larger than
the peak area of acetaminophen from the standard solution.
Acetazolamide contains not less than 98.0z and not
more than 102.0z of acetazolamide (C4H6N4O3S2),
calculated on the dried basis.
length: 225 nm).
Column: A stainless steel column about 4 mm in inside Description Acetazolamide occurs as a white to pale yel-
diameter and about 15 cm in length, packed with octadecyl- lowish white crystalline powder. It is odorless, and has a
silanized silica gel for liquid chromatography (5 mm in parti- slight bitter taste.
cle diameter). It is slightly soluble in ethanol (95), very slightly soluble in
Column temperature: A constant temperature of about water, and practically insoluble in diethyl ether.
409 C. Melting point: about 2559 C (with decomposition).
Mobile phase: A mixture of 0.05 mol/L potassium dihy-
Identification (1) To 0.1 g of Acetazolamide add 5 mL of
drogenphosphate (pH 4.7) and methanol (4:1).
sodium hydroxide TS, then add 5 mL of a solution of 0.1 g
Flow rate: Adjust so that the retention time of acetamino-
of hydroxylammonium chloride and 0.05 g of copper (II)
phen is about 5 minutes.
sulfate pentahydrate in 10 mL of water: a light yellow color
Selection of column: Dissolve 0.01 g each of Acetamino-
develops. Then heat this solution for 5 minutes: a deep yel-
phen and 4-aminophenol hydrochloride in 1 mL of metha-
low color is produced gradually.
nol, add the mobile phase to make 50 mL, to 1 mL of this
(2) To 0.02 g of Acetazolamide add 2 mL of dilute hy-
solution add the mobile phase to make 10 mL. Proceed with
drochloric acid, boil for 10 minutes, cool, and add 8 mL of
10 mL of this solution under the above operating conditions,
water: this solution responds to the Qualitative Tests <1.09>
and calculate the resolution. Use a column giving elution of
for primary aromatic amines.
4-aminophenol and acetaminophen in this order with the
(3) To 0.2 g of Acetazolamide add 0.5 g of granulated
resolution between these peaks being not less than 7.
zinc and 5 mL of diluted hydrochloric acid (1 in 2): the gas
Detection sensitivity: Adjust the detection sensitivity so
evolved darkens moistened lead (II) acetate paper.
that the peak height of acetaminophen obtained from 10 mL
of the standard solution is about 15z of the full scale. Purity (1) Clarity and color of solution—Dissolve 1.0 g
Time span of measurement: About 6 times as long as the of Acetazolamide in 10 mL of sodium hydroxide TS: the
retention time of acetaminophen, beginning after the solvent solution is clear and colorless to pale yellow.
peak. (2) Chloride <1.03>—To 1.5 g of Acetazolamide add 75
mL of water, and warm at 709C for 20 minutes with occa-
sional shaking. After cooling, filter, and to 25 mL of the
2 hours).
Residue on ignition <2.44> Not more than 0.1z (1 g). mL. Perform the test using this solution as the test solution.
Prepare the control solution with 0.20 mL of 0.01 mol/L
Assay Weigh accurately about 20 mg each of Acetamino-
hydrochloric acid VS (not more than 0.014z).
phen and Acetaminophen RS, previously dried, dissolve in
2 mL of methanol, and add water to make exactly 100 mL.
Pipet 3 mL each of these solutions, add water to make ex-
50 mL. Perform the test using this solution as the test solu-
actly 100 mL, and use these solutions as the sample solution
and the standard solution, respectively. Determine the absor-
mol/L sulfuric acid VS (not more than 0.038z).
bances, AT and AS, of the sample solution and standard
(4) Heavy metals <1.07>—Proceed with 1.0 g of Aceta-
solution at the wavelength of maximum absorption at about
zolamide according to Method 2, and perform the test.
244 nm as directed under Ultraviolet-visible Spectropho-
tometry <2.24>, using water as the blank.
Amount (mg) of acetaminophen (C8H9NO2) (5) Silver-reducing substances—Wet 5 g of Acetazola-
＝ M S × AT / AS mide with 5 mL of aldehyde-free ethanol, and add 125 mL
of water, 10 mL of nitric acid and exactly 5 mL of 0.1 mol/L
MS: Amount (mg) of Acetaminophen RS taken
silver nitrate VS. Stir for 30 minutes by protecting from
Containers and storage Containers—Tight containers. light, filter through a glass filter (G3), and wash the residue
Storage—Light-resistant. on the glass filter with two 10-mL portions of water. Com-
bine the filtrate with the washings, to the solution add 5 mL
of ferric ammonium sulface TS, and titrate <2.50> with 0.1
mol/L ammonium thiocyanate VS: not less than 4.8 mL of
0.1 mol/L ammonium thiocyanate VS is consumed.
JP XVII Official Monographs / Glacial Acetic Acid 365

3 hours). Glacial Acetic Acid
Residue on ignition <2.44> Not more than 0.1z (0.5 g).
氷酢酸
Assay Weigh accurately about 0.15 g of Acetazolamide,
and dissolve in 400 mL of water in a water bath by heating.
After cooling, add water to make exactly 1000 mL. Pipet 5
C2H4O2: 60.05
mL of the solution, add 10 mL of 1 mol/L hydrochloric acid
Acetic acid
TS, and then add water to make exactly 100 mL. Determine
[64-19-7]
the absorbance A of this solution at the wavelength of maxi-
mum absorption at about 265 nm as directed under Ultravio-
Glacial Acetic Acid contains not less than 99.0z of
acetic acid (C2H4O2).
Amount (mg) of acetazolamide (C4H6N4O3S2)
Description Glacial Acetic Acid is a clear, colorless, vola-
＝ A/474 × 200,000
tile liquid, or colorless or white, crystalline masses. It has a
Containers and storage Containers—Well-closed contain- pungent, characteristic odor.
ers. It is miscible with water, with ethanol (95) and with diethyl
Storage—Light-resistant. ether.
Boiling point: about 1189 C
Specific gravity d 20
20 : about 1.049
Acetic Acid Identification A solution of Glacial Acetic Acid (1 in 3)
changes blue litmus paper to red, and responds to the
酢酸
Qualitative Tests <1.09> for acetate.
Congealing point <2.42> Not less than 14.59
C.
Acetic Acid contains not less than 30.0 w/vz and
not more than 32.0 w/vz of acetic acid (C2H4O2: Purity (1) Chloride—To 10 mL of Glacial Acetic Acid
60.05). add water to make 100 mL, and use this solution as the sam-
ple solution. To 10 mL of the sample solution add 5 drops of
Description Acetic Acid is a clear, colorless liquid. It has a
silver nitrate TS: no opalescence is produced.
pungent, characteristic odor and an acid taste.
(2) Sulfate—To 10 mL of the sample solution obtained
It is miscible with water, with ethanol (95) and with
in (1) add 1 mL of barium chloride TS: no turbidity is pro-
glycerin.
duced.
20: about 1.04
(3) Heavy metals <1.07>—Evaporate 2.0 mL of Glacial
Identification Acetic Acid changes blue litmus paper to Acetic Acid on a water bath to dryness. Dissolve the residue
red, and responds to the Qualitative Tests <1.09> for acetate. in 2 mL of dilute acetic acid and water to make 50 mL, and
perform the test using this solution as the test solution. Pre-
Purity (1) Chloride—To 20 mL of Acetic Acid add 40 mL
pare the control solution with 2.0 mL of Standard Lead So-
of water, and use this solution as the sample solution. To 10
lution by adding 2.0 mL of dilute acetic acid and water to
mL of the sample solution add 5 drops of silver nitrate TS:
make 50 mL (not more than 10 ppm).
no opalescence is produced.
(4) Potassium permanganate-reducing substances—To
(2) Sulfate—To 10 mL of the sample solution obtained
20 mL of the sample solution obtained in (1) add 0.10 mL of
in (1) add 1 mL of barium chloride TS: no turbidity is pro-
0.1 mol/L potassium permanganate VS: the red color does
duced.
not disappear within 30 minutes.
(3) Heavy metals <1.07>—Evaporate 10 mL of Acetic
(5) Non-volatile residue—Evaporate 10 mL of Glacial
Acid on a water bath to dryness, and to the residue add 2 mL
Acetic Acid on a water bath to dryness, and dry at 1059C for
of dilute acetic acid and water to make 50 mL. Perform the
1 hour: the mass of the residue is not more than 1.0 mg.
test with this solution as the test solution. Prepare the con-
trol solution with 3.0 mL of Standard Lead Solution by add- Assay Place 10 mL of water in a glass-stoppered flask, and
ing 2 mL of dilute acetic acid and water to make 50 mL (not weigh accurately. Add about 1.5 g of Glacial Acetic Acid,
more than 3 ppm). weigh accurately again, then add 30 mL of water, and titrate
(4) Potassium permanganate-reducing substances—To <2.50> with 1 mol/L sodium hydroxide VS (indicator: 2
20 mL of the sample solution obtained in (1) add 0.02 mol/L drops of phenolphthalein TS).
potassium permanganate VS: the red color does not disap-
Each mL of 1 mol/L sodium hydroxide VS
pear within 30 minutes.
＝ 60.05 mg of C2H4O2
(5) Non-volatile residue—Evaporate 30 mL of Acetic
Acid on a water bath to dryness, and dry at 1059C for 1 Containers and storage Containers—Tight containers.
hour: the mass of the residue is not more than 1.0 mg.
Assay Measure exactly 5 mL of Acetic Acid, add 30 mL of
water, and titrate <2.50> with 1 mol/L sodium hydroxide VS
(indicator: 2 drops of phenolphthalein TS).
＝ 60.05 mg of C2H4O2
366 Acetohexamide / Official Monographs JP XVII
(4) Related substances (i) Cyclohexylamine—Dissolve
Acetohexamide exactly 1.0 g of Acetohexamide in exactly 30 mL of 0.5
mol/L sodium hydroxide TS, add exactly 5 mL of hexane,
アセトヘキサミド shake vigorously for 60 minutes, allow to stand for 5
minutes, and use the upper layer as the sample solution.
Separately, dissolve exactly 50 mg of cyclohexylamine in 0.5
mol/L sodium hydroxide TS to make exactly 50 mL. Pipet
2 mL of this solution, and add 0.5 mol/L sodium hydroxide
TS to make exactly 300 mL. Pipet 30 mL of this solution,
add exactly 5 mL of hexane, shake vigorously for 60
minutes, allow to stand for 5 minutes, and use the upper
C15H20N2O4S: 324.40
layer as the standard solution. Perform the test with exactly
4-Acetyl-N-(cyclohexylcarbamoyl)benzenesulfonamide
[968-81-0]
directed under Gas Chromatography <2.02> according to the
following conditions, and determine the peak area of cyclo-
Acetohexamide, when dried, contains not less than
hexylamine by the automatic integration method: the peak
98.0z and not more than 101.0z of acetohexamide
area of cyclohexylamine with the sample solution is not more
(C15H20N2O4S).
than that with the standard solution.
Description Acetohexamide occurs as a white to yellowish Operating conditions—
white powder. Detector: A hydrogen flame-ionization detector.
It is freely soluble in N, N-dimethylformamide, sparingly Column: A fused-silica column 0.53 mm in inside diame-
soluble in acetone, slightly soluble in methanol and in ter and 30 m in length, coated the inner surface with methyl-
ethanol (99.5), and practically insoluble in water. silicone polymer for gas chromatography 1.5 mm in thick-
Melting point: about 1859C (with decomposition). ness.
Identification (1) Dissolve 0.10 g of Acetohexamide in
909C.
100 mL of methanol. To 5 mL of the solution add 20 mL of
Injection port temperature: A constant temperature of
0.5 mol/L hydrochloric acid TS and 75 mL of methanol, and
about 1509C.
use the solution as the sample solution (1). Determine the
Detector temperature: A constant temperature of about
absorption spectrum of the sample solution (1) as directed
2109C.
under Ultraviolet-visible Spectrophotometry <2.24>, using
Carrier gas: Helium.
methanol as the blank, and compare the spectrum with the
Flow rate: Adjust so that the retention time of cyclohex-
Reference Spectrum 1: both spectra exhibit similar intensities
ylamine is about 4 minutes.
of absorption at the same wavelengths. Separately, to exactly
Split ratio: 1:1.
10 mL of the sample solution (1) add methanol to make ex-
actly 50 mL, and use the solution as the sample solution (2).
System performance: When the procedure is run with 2 mL
Determine the absorption spectrum of the sample solution
of the standard solution under the above operating condi-
(2) as directed under Ultraviolet-visible Spectrophotometry
tions, the number of theoretical plates of the peak of cyclo-
<2.24>, using methanol as the blank, and compare the spec-
hexylamine is not less than 8000.
trum with the Reference Spectrum 2: both spectra exhibit
similar intensities of absorption at the same wavelengths.
with 2 mL of the standard solution under the above operating
conditions, the relative standard deviation of the peak area
Acetohexamide, previously dried, as directed in the potas-
of cyclohexylamine is not more than 5z.
sium bromide disk method under Infrared Spectrophotome-
(ii) Dicyclohexylurea—Dissolve exactly 1.0 g of Aceto-
try <2.25>, and compare the spectrum with the Reference
hexamide in exactly 10 mL of 0.5 mol/L sodium hydroxide
Spectrum: both spectra exhibit similar intensities of absorp-
TS, add exactly 20 mL of methanol, shake, then add exactly
tion at the same wave numbers.
5 mL of diluted hydrochloric acid (1 in 10), shake vigorously
Purity (1) Chloride <1.03>—Dissolve 1.5 g of Aceto- for 15 minutes, and centrifuge. Filter 10 mL or more of the
hexamide in 40 mL of N, N-dimethylformamide, add 6 mL supernatant liquid through a membrane filter with pore size
of dilute nitric acid and N, N-dimethylformamide to make 50 of not larger than 0.5 mm. Discard the first 5 mL of the fil-
mL. Perform the test using this solution as the test solution. trate, and use the subsequent filtrate as the sample solution.
Prepare the control solution as follows: to 0.45 mL of 0.01 Separately, dissolve exactly 50 mg of dicyclohexylurea in
mol/L hydrochloric acid VS add 6 mL of dilute nitric acid methanol to make exactly 100 mL. Pipet 2 mL of this solu-
and N, N-dimethylformamide to make 50 mL (not more than tion, and add methanol to make exactly 100 mL. Pipet 20
0.011z). mL of this solution, add exactly 10 mL of 0.5 mol/L sodium
(2) Sulfate <1.14>—Dissolve 2.0 g of Acetohexamide in hydroxide TS, shake, then add exactly 5 mL of diluted hy-
40 mL of N, N-dimethylformamide, and add 1 mL of dilute drochloric acid (1 in 10), shake, and use this solution as the
hydrochloric acid and N, N-dimethylformamide to make 50 standard solution. Perform the test with exactly 50 mL each
mL. Perform the test using this solution as the test solution. of the sample solution and standard solution as directed
Prepare the control solution as follows: to 0.40 mL of 0.005 under Liquid Chromatography <2.01> according to the fol-
mol/L sulfuric acid VS add 1 mL of dilute hydrochloric acid lowing conditions, and determine the peak area of dicyclo-
and N, N-dimethylformamide to make 50 mL (not more than hexylurea by the automatic integration method: the peak
0.010z). area of dicyclohexylurea with the sample solution is not
(3) Heavy metals <1.07>—Proceed with 1.0 g of Aceto- more than that with the standard solution.
hexamide according to Method 2, and perform the test. Operating conditions—
Prepare the control solution with 2.0 mL of Standard Lead Detector: An ultraviolet absorption photometer (wave-
Solution (not more than 20 ppm). length: 210 nm).
JP XVII Official Monographs / Acetylcholine Chloride for Injection 367

ter and 25 cm in length, packed with octadecylsilanized silica Acetylcholine Chloride for Injection
Column temperature: A constant temperature of about 注射用アセチルコリン塩化物
259 C.
Mobile phase: Dissolve 0.5 g of sodium hydroxide in 1000
mL of 0.05 mol/L sodium dihydrogen phosphate TS, and
adjust the pH to 6.5 with 0.5 mol/L sodium hydroxide TS.
To 500 mL of this solution add 500 mL of acetonitrile. C7H16ClNO2: 181.66
Flow rate: Adjust so that the retention time of dicyclo- 2-Acetoxy-N, N, N-trimethylethylaminium chloride
hexylurea is about 10 minutes. [60-31-1]
System performance: When the procedure is run with 50 Acetylcholine Chloride for Injection is a prepara-
mL of the standard solution under the above operating tion for injection which is dissolved before use.
conditions, the number of theoretical plates of the peak of It contains not less than 98.0z and not more than
dicyclohexylurea is not less than 10,000. 102.0z of acetylcholine chloride (C7H16ClNO2), and
System repeatability: When the test is repeated 6 times not less than 19.3z and not more than 19.8z of chlo-
with 50 mL of the standard solution under the above operat- rine (Cl: 35.45), calculated on the dried basis.
ing conditions, the relative standard deviation of the peak It contains not less than 93.0z and not more than
area of dicyclohexylurea is not more than 2.0z. 107.0z of the labeled amount of acetylcholine chlo-
(iii) Other related substances—Dissolve 0.10 g of Aceto- ride (C7H16ClNO2).
hexamide in 10 mL of acetone, and use this solution as the
Method of preparation Prepare as directed under Injec-
sample solution. Pipet 1 mL of the sample solution, and add
tions.
acetone to make exactly 20 mL. Pipet two 1 mL portions of
this solution, add acetone to make exactly 10 mL and 25 mL, Description Acetylcholine Chloride for Injection occurs as
respectively, and use these solutions as the standard solution white, crystals or crystalline powder.
(1) and the standard solution (2). Perform the test with these It is very soluble in water, and freely soluble in ethanol
solutions as directed under Thin-layer Chromatography (95).
<2.03>. Spot 10 mL each of the sample solution and standard It is extremely hygroscopic.
solutions (1) and (2) on a plate of silica gel with fluorescent
Identification (1) Determine the infrared absorption spec-
indicator for thin-layer chromatography. Develop the plate
trum of Acetylcholine Chloride for Injection, previously
with a mixture of ethyl acetate, methanol, ammonia solution
dried, as directed in the potassium bromide disk method
(28) and cyclohexane (6:2:1:1) to a distance of about 10 cm,
under Infrared Spectrophotometry <2.25>, and compare the
and air-dry the plate. Examine under ultraviolet light (main
spectrum with the Reference Spectrum: both spectra exhibit
wavelength: 254 nm): the spots other than the principal spot
similar intensities of absorption at the same wave numbers.
from the sample solution are not more intense than spot
(2) A solution of Acetylcholine Chloride for Injection
from the standard solution (1), and the number of them
(1 in 10) responds to the Qualitative Tests <1.09> (2) for
which are more intense than the spot from the standard
chloride.
solution (2) is not more than 4.
Melting point <2.60> 149 – 1529C. Seal Acetylcholine Chlo-
Loss on drying <2.41> Not more than 1.0z (1 g, 1059C,
ride for Injection in a capillary tube for melting point imme-
4 hours).
diately after drying both of the sample and the tube at 1059C
Residue on ignition <2.44> Not more than 0.1z (1 g). for 3 hours, and determine the melting point.
Assay Weigh accurately about 0.3 g of Acetohexamide, Purity (1) Clarity and color of solution—Dissolve 1.0 g
previously dried, dissolve in 30 mL of N, N-dimethylfor- of Acetylcholine Chloride for Injection in 10 mL of water:
mamide, add 10 mL of water, and titrate <2.50> with 0.1 the solution is clear and colorless.
mol/L sodium hydroxide VS (potentiometric titration). (2) Acidity—Dissolve 0.10 g of Acetylcholine Chloride
Perform a blank determination using a solution prepared for Injection in 10 mL of freshly boiled and cooled water,
by adding 19 mL of water to 30 mL of N, N-dimethylfor- and add 1 drop of bromothymol blue TS, and 0.30 mL of
mamide, and make any necessary correction. 0.01 mol/L sodium hydroxide VS: the solution is blue in
color.
(3) Heavy metals <1.07>—Proceed with 2.0 g of Acetyl-
＝ 32.44 mg of C15H20N2O4S
choline Chloride for Injection according to Method 1, and
Containers and storage Containers—Well-closed contain- perform the test. Prepare the control solution with 2.0 mL of
ers. Standard Lead Solution (not more than 10 ppm).
3 hours).
Uniformity of dosage units <6.02> It meets the requirement
of the Mass variation test.
Foreign insoluble matter <6.06> Perform the test according
to Method 2: it meets the requirement.
Insoluble particulate matter <6.07> It meets the require-
ment.
368 Acetylcysteine / Official Monographs JP XVII
Sterility <4.06> Perform the test according to the Mem- and cool. Then add 5 mL of nitric acid, add water to make
brane filtration method: it meets the requirement. 50 mL, and perform the test using this solution as the test so-
lution. Prepare the control solution with 0.45 mL of 0.01
Assay (1) Acetylcholine chloride—Weigh accurately the
contents of not less than 10 Acetylcholine Chloride for Injec-
(2) Sulfate <1.14>—Perform the test with 0.8 g of Acetyl-
tions. Weigh accurately about 0.5 g of the contents, dissolve
cysteine. Prepare the control solution with 0.50 mL of 0.005
in 15 mL of water, then add exactly 40 mL of 0.1 mol/L so-
dium hydroxide VS, stopper loosely, and heat on a water
(3) Ammonium <1.02>—Perform the test with 0.10 g of
bath for 30 minutes. Cool quickly, and titrate <2.50> the
Acetylcysteine, using the distillation under reduced pressure.
excess sodium hydroxide with 0.05 mol/L sulfuric acid VS
Prepare the control solution with 2.0 mL of Standard
(indicator: 3 drops of phenolphthalein TS). Perform a blank
Ammonium Solution (not more than 0.02z).
determination.
(4) Heavy metals <1.07>—Dissolve 1.0 g of Acetyl-
Each mL of 0.1 mol/L sodium hydroxide VS cysteine in 40 mL of water, add 3 mL of sodium hydroxide
＝ 18.17 mg of C7H16ClNO2 TS, 2 mL of dilute acetic acid and water to make 50 mL, and
(2) Chlorine—Titrate <2.50> the solution, which has been
pare the control solution as follows: To 1.0 mL of Standard
titrated in (1), with 0.1 mol/L silver nitrate VS (indicator:
Lead Solution add 2 mL of dilute acetic acid and water to
3 drops of fluorescein sodium TS).
Each mL of 0.1 mol/L silver nitrate VS (5) Iron <1.10>—Prepare the test solution with 1.0 g of
＝ 3.545 mg of Cl Acetylcysteine according to Method 1, and perform the test
according to Method A. Prepare the control solution with
Containers and storage Containers—Hermetic containers.
1.0 mL of Standard Iron Solution (not more than 10 ppm).
(6) Related substances—Dissolve 50 mg of Acetyl-
cysteine in 25 mL of the mobile phase, and use this solution
Acetylcysteine as the sample solution. The sample solution is prepared
before using. Perform the test with 10 mL of the sample
アセチルシステイン
solution as directed under Liquid Chromatography <2.01>
according to the following conditions, determine each peak
area by the automatic integration method, and calculate
their amounts by the area percentage method: the area of the
peak other than acetylcysteine is not more than 0.3z, and
the total area of the peak other than acetylcysteine is not
C5H9NO3S: 163.19 more than 0.6z.
(2R)-2-Acetylamino-3-sulfanylpropanoic acid Operating conditions—
[616-91-1] Detector: An ultraviolet absorption photometer (wave-
length: 220 nm).
Acetylcysteine contains not less than 99.0z and not Column: A stainless steel column 4.6 mm in inside diame-
more than 101.0z of acetylcysteine (C5H9NO3S), cal- ter and 25 cm in length, packed with octadecylsilanized silica
culated on the dried basis. gel for liquid chromatography (5 mm in particle diameter).
Description Acetylcysteine occurs as white, crystals or crys-
409C.
talline powder.
Mobile phase: A mixture of diluted phosphoric acid (1 in
It is freely soluble in water and in ethanol (99.5).
2500) and acetonitrile (19:1).
Flow rate: Adjust so that the retention time of acetyl-
Identification Determine the infrared absorption spectrum cysteine is about 7 minutes.
of Acetylcysteine as directed in the potassium bromide disk Time span of measurement: About 4 times as long as the
method under Infrared Spectrophotometry <2.25>, and com- retention time of acetylcysteine, beginning after the solvent
pare the spectrum with the Reference Spectrum: both spectra peak.
exhibit similar intensities of absorption at the same wave System suitability—
numbers. Test for required detectability: To 1 mL of the sample so-
lution add the mobile phase to make 10 mL. To 1 mL of this
Optical rotation <2.49> [a]20D : ＋21.0 – ＋27.09Weigh accu-
solution, add the mobile phase to make 20 mL, and use this
rately an amount of Acetylcysteine, equivalent to about 2.5 g
solution as the solution for system suitability test. Pipet 5
calculated on the dried basis, and dissolve with 2 mL of a so-
mL of the solution for system suitability test, and add the
lution of disodium dihydrogen ethylenediamine tetraacetate
mobile phase to make exactly 25 mL. Confirm that the peak
dihydrate (1 in 100) and 15 mL of a solution of sodium hy-
area of acetylcysteine obtained with 10 mL of this solution is
droxide (1 in 25). To this solution add a solution, prepared
equivalent to 15 to 25z of that obtained with 10 mL of the
by adjusting the pH to 7.0 of 500 mL of a solution of potas-
solution for system suitability test.
sium dihydrogen phosphate (17 in 125) with sodium hydrox-
System performance: When the procedure is run with 10
ide TS and adding water to make 1000 mL, to make exactly
mL of the solution for system suitability test under the above
50 mL. Determine the optical rotation of this solution using
operating conditions, the number of theoretical plates and
a 100-mm cell.
the symmetry factor of the peak of acetylcysteine are not less
Melting point <2.60> 107 – 1119C than 15,000 and not more than 1.5, respectively.
Purity (1) Chloride <1.03>—Dissolve 0.40 g of Acetyl-
with 10 mL of the solution for system suitability test under
cysteine in 25 mL of sodium hydroxide TS, add 4 mL of
the above operating conditions, the relative standard devia-
hydrogen peroxide (30), heat in a water bath for 45 minutes,
tion of the peak area of acetylcysteine is not more than
JP XVII Official Monographs / Aciclovir 369
2.0z. more than 10 ppm).

(3) Related substances—Use the sample solution ob-
tained in the Assay as the sample solution. Separately, weigh
3 hours).
accurately about 25 mg of guanine, dissolve in 50 mL of
Residue on ignition <2.44> Not more than 0.3z (1 g). dilute sodium hydroxide TS, and add the mobile phase to
make exactly 100 mL. Pipet 2 mL of this solution, add the
Assay Weigh accurately about 0.2 g of Acetylcysteine,
mobile phase to make exactly 100 mL, and use this solution
place it in a stoppered flask, dissolve in 20 mL of water, add
as the standard solution. Perform the test with exactly 10 mL
4 g of potassium iodide and 5 mL of dilute hydrochloric
each of the sample solution and standard solution as directed
acid, then add exactly 25 mL of 0.05 mol/L iodine VS, stop-
under Liquid Chromatography <2.01> according to the fol-
per tightly, allow to stand for 20 minutes in an ice cold water
lowing conditions. Determine the peak areas of guanine, AT
in the dark, and titrate <2.50> the excess of iodine with 0.1
and AS, and calculate the amount of guanine by the follow-
mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS).
ing equation: it is not more than 0.7z. Determine each peak
Perform a blank determination in the same manner.
area from the sample solution by the automatic integration
Each mL of 0.05 mol/L iodine VS method, and calculate the amount of each related substance
＝ 16.32 mg of C5H9NO3S other than aciclovir and guanine by the area percentage
method: it is not more than 0.2z. Furthermore, the sum of
the amount of guanine calculated above and the amounts of
related substances determined by the area percentage method
is not more than 1.5z.
Aciclovir
Amount (z) of guanine ＝ MS/MT × AT/AS × 2/5
アシクロビル
MS: Amount (mg) of guanine taken
MT: Amount (mg) of Aciclovir taken
Detector, column, column temperature, mobile phase, and
flow rate: Proceed as directed in the operating conditions in
the Assay.
C8H11N5O3: 225.20 Time span of measurement: About 8 times as long as the
2-Amino-9-[(2-hydroxyethoxy)methyl]-1,9-dihydro- retention time of aciclovir, beginning after the solvent peak.
6H-purin-6-one System suitability—
[59277-89-3] System performance: Proceed as directed in the system
suitability in the Assay.
Aciclovir contains not less than 98.5z and not more Test for required detectability: To 1 mL of the sample
than 101.0z of aciclovir (C8H11N5O3), calculated on solution add the mobile phase to make 100 mL, and use this
the anhydrous basis. solution as the solution for system suitability test. Pipet 1
Description Aciclovir occurs as a white to pale yellowish
white crystalline powder.
area of aciclovir obtained from 10 mL of this solution is
It is slightly soluble in water and very slightly soluble in
equivalent to 7 to 13z of that obtained from 10 mL of the
ethanol (99.5).
It dissolves in 0.1 mol/L hydrochloric acid TS and in
dilute sodium hydroxide TS.
with 10 mL of the standard solution under the above operat-
Identification (1) Determine the absorption spectrum of a ing conditions, the relative standard deviation of the peak
solution of Aciclovir in 0.1 mol/L hydrochloric acid TS (1 in area of guanine is not more than 2.0z.
100,000) as directed under Ultraviolet-visible Spectropho-
Water <2.48> Not more than 6.0z (50 mg, coulometric
tometry <2.24>, and compare the spectrum with the Refer-
titration).
ence Spectrum or the spectrum of a solution of Aciclovir RS
prepared in the same manner as the sample solution: both Residue on ignition <2.44> Not more than 0.1z (1 g).
Assay Weigh accurately about 20 mg each of Aciclovir and
wavelengths.
Aciclovir RS (separately determine the water <2.48> in the
same manner as Aciclovir), dissolve each in 1 mL of dilute
Aciclovir as directed in the potassium bromide disk method
sodium hydroxide TS, add the mobile phase to make exactly
20 mL each, and use these solutions as the sample solution
spectrum with the Reference Spectrum or the spectrum of
and the standard solution, respectively. Perform the test
Aciclovir RS: both spectra exhibit similar intensities of ab-
with exactly 10 mL each of the sample solution and standard
sorption at the same wave numbers.
Purity (1) Clarity and color of solution—Dissolve 0.5 g according to the following conditions, and determine the
of Aciclovir in 20 mL of dilute sodium hydroxide TS: the so- peak areas, AT and AS, of aciclovir in each solution.
lution is clear and is not more colored than the following
Amount (mg) of aciclovir (C8H11N5O3) ＝ MS × AT/AS
control solution.
Control solution: To 2.5 mL of Matching Fluid F add MS: Amount (mg) of Aciclovir RS taken, calculated on
diluted dilute hydrochloric acid (1 in 10) to make 100 mL. the anhydrous basis
(2) Heavy metals <1.07>—Proceed with 1.0 g of Aciclovir
according to Method 2, and perform the test. Prepare the
control solution with 1.0 mL of Standard Lead Solution (not
370 Aciclovir Granules / Official Monographs JP XVII
length: 254 nm). Dissolution <6.10> When the test is performed at 50 revolu-
Column: A stainless steel column 4.6 mm in inside diame- tions per minute according to the Paddle method, using 900
ter and 10 cm in length, packed with octadecylsilanized silica mL of water as the dissolution medium, the dissolution rate
gel for liquid chromatography (3 mm in particle diameter). in 30 minutes of Aciclovir Granules is not less than 85z.
Column temperature: A constant temperature of about Start the test with an accurately weighed amount of
209 C. Aciclovir Granules, equivalent to about 0.4 g of aciclovir
Mobile phase: Dissolve 1.0 g of sodium 1-decanesulfonate (C8H11N5O3), withdraw not less than 20 mL of the medium
and 6.0 g of sodium dihydrogen phosphate dihydrate in 1000 at the specified minute after starting the test, and filter
mL of water, and adjust the pH to 3.0 with phosphoric acid. through a membrane filter with a pore size not exceeding
To this solution add 40 mL of acetonitrile. 0.45 mm. Discard the first 10 mL of the filtrate, pipet 2 mL
Flow rate: Adjust so that the retention time of aciclovir is of the subsequent filtrate, add water to make exactly 100
about 3 minutes. mL, and use this solution as the sample solution. Separately,
System suitability— weigh accurately about 22 mg of Aciclovir RS (separately
System performance: Dissolve 0.1 g of Aciclovir in 5 mL determine the water <2.48> in the same manner as Aciclovir),
of dilute sodium hydroxide TS, add 2 mL of a solution of and dissolve in water to make exactly 100 mL. Pipet 4 mL of
guanine in dilute sodium hydroxide TS (1 in 4000), and add this solution, add water to make exactly 100 mL, and use
the mobile phase to make 100 mL. When the procedure is this solution as the standard solution. Determine the absor-
run with 10 mL of this solution under the above operating bances, AT and AS, at 252 nm of the sample solution and
conditions, aciclovir and guanine are eluted in this order standard solution as directed under Ultraviolet-visible
with the resolution between these peaks being not less than Spectrophotometry <2.24>.
17.
of aciclovir (C8H11N5O3)
＝ MS/MT × AT/AS × 1/C × 1800
ing conditions, the relative standard deviation of the peak
area of aciclovir is not more than 1.0z. MS: Amount (mg) of aciclovir RS taken, calculated on the
anhydrous basis
Containers and storage Containers—Well-closed contain-
MT: Amount (g) of Aciclovir Granules taken
ers.
C: Labeled amount (mg) of aciclovir (C8H11N5O3) in 1 g
Assay Powder Aciclovir Granules, and weigh accurately a
Aciclovir Granules portion of the powder, equivalent to about 0.1 g of aciclovir
(C8H11N5O3), add 60 mL of dilute sodium hydroxide TS,
アシクロビル顆粒 agitate with the aid of ultrasonic waves for 15 minutes, then
add dilute sodium hydroxide TS to make exactly 100 mL,
and filter. Discard the first 20 mL of filtrate, pipet 15 mL of
Aciclovir Granules contain not less than 93.0z
the subsequent filtrate, add 50 mL of water and 5.8 mL of 2
and not more than 107.0z of the labeled amount of
mol/L hydrochloric acid TS, and add water to make exactly
aciclovir (C8H11N5O3: 225.20).
100 mL. Pipet 5 mL of this solution, add 0.1 mol/L hydro-
Method of preparation Prepare as directed under Gran- chloric acid TS to make exactly 100 mL, and use this solu-
ules, with Aciclovir. tion as the sample solution. Separately, weigh accurately
about 25 mg of Aciclovir RS (separately determine the water
Identification Determine the absorption spectrum of the
<2.48> in the same manner as Aciclovir), and dissolve in
sample solution obtained in the Assay as directed under
dilute sodium hydroxide TS to make exactly 25 mL. Pipet 15
Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a
mL of this solution, add 50 mL of water and 5.8 mL of 2
maximum between 254 nm and 258 nm.
mol/L hydrochloric acid TS, add water to make exactly 100
Uniformity of dosage units <6.02> Perform the test ac- mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric
cording to the following method: Aciclovir Granules in acid TS to make exactly 100 mL, and use this solution as the
single-dose packages meet the requirement of the Content standard solution. Determine the absorbances, AT and AS, at
uniformity test. 255 nm of the sample solution and standard solution as
To the total amount of the content of 1 package of directed under Ultraviolet-visible Spectrophotometry <2.24>,
Aciclovir Granules, add 100 mL of dilute sodium hydroxide using 0.1 mol/L hydrochloric acid TS as the blank.
TS, agitate with the aid of ultrasonic waves with occasional
Amount (mg) of acyclovir (C8H11N5O3)
shaking, and add dilute sodium hydroxide TS to make ex-
＝ M S × AT / AS × 4
actly 200 mL. Filter this solution, discard the first 20 mL of
the filtrate, pipet V mL of the subsequent filtrate, add dilute MS: Amount (mg) of Aciclovir RS taken, calculated on
sodium hydroxide TS to make exactly V? mL so that each the anhydrous basis
mL contains about 1 mg of aciclovir (C8H11N5O3). Pipet 15
mL of this solution, add 50 mL of water and 5.8 mL of 2
mol/L hydrochloric acid TS, add water to make exactly 100
mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric
acid TS to make exactly 100 mL, and use this solution as the
sample solution. Then, proceed as directed in the Assay.
＝ MS × AT/AS × V?/V × 8
MS: Amount (mg) of Aciclovir RS taken, calculated on
the anhydrous basis
JP XVII Official Monographs / Aciclovir for Injection 371
dilute acetic acid add water to make 900 mL. Adjust this so-
Aciclovir Injection lution to pH 2.5 with 1 mol/L sodium hydroxide TS, and
add water to make 1000 mL. To 950 mL of this solution add
アシクロビル注射液 50 mL of methanol.
Flow rate: Adjust so that the retention time of aciclovir is
about 7 minutes.
Aciclovir Injection is an aqueous injection.
It contains not less than 95.0z and not more
than 105.0z of the labeled amount of aciclovir
mL of the standard solution under the above operating con-
(C8H11N5O3: 225.20).
ditions, the internal standard and aciclovir are eluted in this
Method of preparation Prepare as directed under Injec- order with the resolution between these peaks being not less
tions, with Aciclovir. than 3.
Description Aciclovir Injection occurs as a colorless or pale
yellow, clear liquid.
ing conditions, the relative standard deviation of the ratio of
Identification To a volume of Aciclovir Injection, equiva- the peak area of aciclovir to that of the internal standard is
lent to 25 mg of Aciclovir, add 0.5 mol/L hydrochloric acid not more than 1.0z.
TS to make 100 mL. To 2 mL of this solution add 0.5 mol/L
hydrochloric acid TS to make 50 mL. Determine the absorp-
tion spectrum of this solution as directed under Ultraviolet-
visible Spectrophotometry <2.24>: it exhibits a maximum be-
tween 254 nm and 258 nm. Aciclovir for Injection
Bacterial endotoxins <4.01> Less than 0.5 EU/mg. 注射用アシクロビル
Extractable volume <6.05> It meets the requirement.
Aciclovir for Injection is a preparation for injection
which is dissolved before use.
Insoluble particulate matter <6.07> It meets the require- than 105.0z of the labeled amount of aciclovir
ment. (C8H11N5O3: 225.20).
Sterility <4.06> Perform the test according to the Mem- Method of preparation Prepare as directed under Injec-
brane filtration method: it meets the requirement. tions, with Aciclovir.
Assay To an exact volume of Aciclovir Injection, equiva- Description Aciclovir for Injection occurs as white to pale
lent to about 25 mg of aciclovir (C8H11N5O3), add 0.1 mol/L yellowish white, light masses or powder.
hydrochloric acid TS to make exactly 100 mL. Pipet 2 mL of
this solution, add exactly 5 mL of the internal standard solu-
tion, then add 0.1 mol/L hydrochloric acid TS to make 50
mL, and use this solution as the sample solution. Separately,
weigh accurately about 25 mg of Aciclovir RS (separately
determine the water <2.48> in the same manner as Aciclovir), pH Being specified separately when the drug is granted
and dissolve in 0.1 mol/L hydrochloric acid TS to make approval based on the Law.
exactly 100 mL. Pipet 2 mL of this solution, add exactly
Purity Clarity and color of solution—Dissolve an amount
5 mL of the internal standard solution, then add 0.1 mol/L
of Aciclovir for Injection, equivalent to 0.25 g of Aciclovir,
hydrochloric acid TS to make 50 mL, and use this solution
in 10 mL of water: the solution is clear and is not more
as the standard solution. Perform the test with 20 mL each of
colored than the following control solution.
the sample solution and standard solution as directed under
Control solution: To 2.5 mL of Matching Fluid F add
Liquid Chromatography <2.01> according to the following
diluted dilute hydrochloric acid (1 in 10) to make 100 mL.
conditions, and calculate the ratios, QT and QS, of the peak
area of aciclovir to that of the internal standard. Water <2.48> Not more than 7.5z (0.1 g, volumetric titra-
tion, direct titration).
Amount (mg) of aciclovir (C8H11N5O3)
＝ M S × Q T / QS Bacterial endotoxins <4.01> Less than 0.25 EU/mg.
MS: Amount (mg) of Aciclovir RS taken, calculated on Uniformity of dosage units <6.02> It meets the requirement
the anhydrous basis of the Mass variation test.
Internal standard solution—A solution of nicotinic acid in Foreign insoluble matter <6.06> Perform the test according
0.1 mol/L hydrochloric acid TS (3 in 20,000). to Method 2: it meets the requirement.
ment.
length: 254 nm).
Column: A stainless steel column 4.6 mm in inside diame- Sterility <4.06> Perform the test according to the Mem-
ter and 15 cm in length, packed with octadecylsilanized silica brane filtration method: it meets the requirement.
Assay Weigh accurately the mass of the contents of not
less than 10 Aciclovir for Injection. Weigh accurately an
259 C.
amount of the contents, equivalent to about 0.1 g of
Mobile phase: To 1.45 g of phosphoric acid and 25 mL of
aciclovir (C8H11N5O3), and dissolve in dilute sodium hy-
372 Aciclovir Ointment / Official Monographs JP XVII
droxide TS to make exactly 100 mL. Pipet 15 mL of this
solution, add 70 mL of water and 5 mL of 2 mol/L hy- Aciclovir Ophthalmic Ointment
drochloric acid TS, then add water to make exactly 100 mL.
Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric アシクロビル眼軟膏
Aciclovir Ophthalmic Ointment contains not less
of Aciclovir RS (separately determine the water <2.48> in the
than 90.0z and not more than 110.0z of the labeled
same manner as Aciclovir), and dissolve in dilute sodium
amount of aciclovir (C8H11N5O3: 225.20).
hydroxide TS to make exactly 20 mL. Pipet 15 mL of this
solution, add 70 mL of water and 5 mL of 2 mol/L hydro- Method of preparation Prepare as directed under Oph-
chloric acid TS, then add water to make exactly 100 mL. thalmic Ointments, with Aciclovir.
Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric
standard solution. Determine the absorbances, AT and AS,
at 255 nm of the sample solution and standard solution as
directed under Ultraviolet-visible Spectrophotometry <2.24>,
using 0.1 mol/L hydrochloric acid TS as the blank. Metal Particles <6.01> It meets the requirement.
Amount (mg) of aciclovir (C8H11N5O3) Sterility <4.06> Perform the test according to the Mem-
＝ M S × A T / AS × 5 brane filtration method: it meets the requirement.
MS: Amount (mg) of Aciclovir RS taken, calculated on Particle diameter Being specified separately when the drug
the anhydrous basis is granted approval based on the Law.
Containers and storage Containers—Hermetic containers. Assay Weigh accurately a portion of Aciclovir Ophthalmic
Ointment, equivalent to about 15 mg of aciclovir
(C8H11N5O3), add exactly 20 mL of hexane and exactly 20
Aciclovir Ointment mL of dilute sodium hydroxide TS, and shake vigorously.
Centrifuge this mixture, discard the upper layer, pipet 1 mL
アシクロビル軟膏 of the lower layer, add 70 mL of water and 5 mL of 2 mol/L
hydrochloric acid TS, then add water to make exactly 100
Aciclovir Ointment contains not less than 95.0z
weigh accurately about 15 mg of Aciclovir RS (separately
determine the water <2.48> in the same manner as Aciclovir),
aciclovir (C8H11N5O3: 225.20).
and dissolve in dilute sodium hydroxide TS to make exactly
Method of preparation Prepare as directed under Oint- 20 mL. Pipet 1 mL of this solution, add 70 mL of water and
ments, with Aciclovir. 5 mL of 2 mol/L hydrochloric acid TS, then add water to
make exactly 100 mL, and use this solution as the standard
solution. Determine the absorbances, AT and AS, at 255 nm
of the sample solution and standard solution as directed
water as the blank.
Assay Weigh accurately an amount of Aciclovir Ointment,
Amount (mg) of acyclovir (C8H11N5O3) ＝ MS × AT/AS
equivalent to about 10 mg of aciclovir (C8H11N5O3), add
25 mL of dilute sodium hydroxide TS, warm if necessary, MS: Amount (mg) of Aciclovir RS taken, calculated on
and dissolve by shaking. After cooling, add water to make the anhydrous basis
exactly 100 mL. Pipet 15 mL of this solution, add 0.1 mol/L
hydrochloric acid TS to make exactly 200 mL, and use this
solution as the sample solution. Separately, weigh accurately
about 20 mg of Aciclovir RS (separately, determine the
water <2.48> in the same manner as Aciclovir), and dissolve Aciclovir Syrup
in dilute sodium hydroxide TS to make exactly 20 mL. Pipet
アシクロビルシロップ
10 mL of this solution, and add 15 mL of dilute sodium
hydroxide TS and water to make exactly 100 mL. Pipet 15
mL of this solution, add 0.1 mol/L hydrochloric acid TS to Aciclovir Syrup is a suspension syrup.
make exactly 200 mL, and use this solution as the standard It contains not less than 93.0z and not more
solution. Determine the absorbances, AT and AS, at 255 nm than 107.0z of the labeled amount of aciclovir
of the sample solution and standard solution as directed (C8H11N5O3: 225.20).
Method of preparation Prepare as directed under Syrups,
0.1 mol/L hydrochloric acid TS as the blank.
with Aciclovir.
Identification To a volume of thoroughly shaked Aciclovir
＝ MS × AT/AS × 1/2
Syrup, equivalent to 80 mg of Aciclovir, add 0.1 mol/L
MS: Amount (mg) of Aciclovir RS taken, calculated on hydrochloric acid TS to make 100 mL. Centrifuge this
the anhydrous basis solution, to 1 mL of the supernatant liquid add 0.1 mol/L
hydrochloric acid TS to make 100 mL. Determine the
absorption spectrum of this solution as directed under
JP XVII Official Monographs / Aciclovir for Syrup 373
maximum between 254 nm and 258 nm. Flow rate: Adjust so that the retention time of aciclovir is
about 7 minutes.
tions per minute according to the Paddle method, using 900
mL of water as the dissolution medium, the dissolution rate
in 15 minutes of Aciclovir Syrup is not less than 85z.
ditions, the internal standard and aciclovir are eluted in this
Start the test with an exact volume of thoroughly shaked
order with the resolution between these peaks being not less
Aciclovir Syrup, equivalent to about 0.4 g of aciclovir
than 3.
(C8H11N5O3), withdraw not less than 20 mL of the medium
at the specified minute after starting the test, and centrifuge.
Pipet 2 mL of the supernatant liquid, add water to make
exactly 100 mL, and use this solution as the sample solution.
the peak area of aciclovir to that of the internal standard is
Separately, weigh accurately about 22 mg of Aciclovir RS
not more than 1.0z.
(separately determine the water <2.48> in the same manner as
Aciclovir), and dissolve in water to make exactly 100 mL. Containers and storage Containers—Tight containers.
Pipet 4 mL of this solution, add water to make exactly 100
mine the absorbances, AT and AS, at 252 nm of the sample Aciclovir for Syrup
solution and standard solution as directed under Ultraviolet-
visible Spectrophotometry <2.24>. シロップ用アシクロビル
of aciclovir (C8H11N5O3) Aciclovir for Syrup is a preparation for syrup,
＝ MS/VT × AT/AS × 1/C × 1800 which is suspended before use.
than 105.0z of the labeled amount of aciclovir
the anhydrous basis
(C8H11N5O3: 225.20).
VT: Amount (mL) of Aciclovir Syrup taken
C: Labeled amount (mg) of aciclovir (C8H11N5O3) in 1 mL Method of preparation Prepare as directed under Prepara-
tions for Syrup, with Aciclovir.
Assay Shake thoroughly Aciclovir Syrup. To an exact
volume of the syrup, equivalent to about 80 mg of aciclovir Identification Dissolve an amount of Aciclovir for Syrup,
(C8H11N5O3), add 0.1 mol/L hydrochloric acid TS to make equivalent to 12 mg of Aciclovir, in 0.1 mol/L hydrochloric
exactly 100 mL. Centrifuge this solution, pipet 2 mL of the acid TS to make 50 mL. To 2 mL of this solution add 0.1
supernatant liquid, add exactly 5 mL of the internal standard mol/L hydrochloric acid TS to make 50 mL, and determine
solution, then add 0.1 mol/L hydrochloric acid TS to make the absorption spectrum of this solution as directed under
50 mL, and use this solution as the sample solution. Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a
Separately, weigh accurately about 40 mg of Aciclovir RS maximum between 254 nm and 258 nm.
Aciclovir), and dissolve in 0.1 mol/L hydrochloric acid TS
ing to the following method: Aciclovir for Syrup in single-
to make exactly 50 mL. Pipet 2 mL of this solution, add
dose packages meets the requirement of the Content unifor-
exactly 5 mL of the internal standard solution, then add 0.1
mity test.
mol/L hydrochloric acid TS to make 50 mL, and use this so-
To the total content of 1 package of Aciclovir for Syrup
lution as the standard solution. Perform the test with 20 mL
add 2V/25 mL of diluted sodium hydroxide TS (1 in 10),
and treat with ultrasonic waves to disintegrate, add water to
make exactly V mL so that each mL contains about 0.8 mg
lowing conditions, and calculate the ratios, QT and QS, of
of aciclovir (C8H11N5O3), and filter this solution through a
the peak area of aciclovir to that of the internal standard.
membrane filter with a pore size not exceeding 0.45 mm.
Amount (mg) of aciclovir (C8H11N5O3) Discard the first 2 mL of the filtrate, pipet 5 mL of the sub-
＝ M S × Q T / QS × 2 sequent filtrate, add exactly 5 mL of the internal standard
solution, then add the mobile phase to make 50 mL, and use
this solution as the sample solution. Then, proceed as di-
the anhydrous basis
rected in the Assay.
Internal standard solution—A solution of nicotinic acid in
0.1 mol/L hydrochloric acid TS (1 in 2000).
＝ MS × QT/QS × V/10
Detector: An ultraviolet absorption photometer (wave- MS: Amount (mg) of Aciclovir RS taken, calculated on
length: 254 nm). the anhydrous basis
Internal standard solution—A solution of parahydroxyben-
zoic acid in the mobile phase (1 in 1250).
Column temperature: A constant temperature of about Dissolution <6.10> When the test is performed at 50 revolu-
259 C. tions per minute according to the Paddle method, using 900
Mobile phase: To 1.45 g of phosphoric acid and 25 mL of mL of water as the dissolution medium, the dissolution rate
dilute acetic acid add water to make 900 mL. Adjust this so- in 45 minutes of Aciclovir for Syrup is not less than 85z.
lution to pH 2.5 with 1 mol/L sodium hydroxide TS, and Start the test with an accurately weighed amount of
add water to make 1000 mL. To 950 mL of this solution add Aciclovir for Syrup, equivalent to about 0.2 g of aciclovir
50 mL of methanol. (C8H11N5O3), withdraw not less than 5 mL of the medium at
374 Aciclovir Tablets / Official Monographs JP XVII
the specified minute after starting the test, and filter through order with the resolution between these peaks being not less
a membrane filter with a pore size not exceeding 0.45 mm. than 20.
Discard the first 2 mL of the filtrate, pipet 2 mL of the sub- System repeatability: When the test is repeated 6 times
sequent filtrate, add water to make exactly 50 mL, and use with 20 mL of the standard solution under the above operat-
this solution as the sample solution. Separately, weigh accu- ing conditions, the relative standard deviation of the ratio of
rately about 11 mg of Aciclovir RS (separately determine the the peak area of aciclovir to that of the internal standard is
water <2.48> in the same manner as Aciclovir), and dissolve not more than 1.0z.
in water to make exactly 50 mL. Pipet 2 mL of this solution,
add water to make exactly 50 mL, and use this solution as
the standard solution. Determine the absorbances, AT and
AS, at 254 nm of the sample solution and standard solution
as directed under Ultraviolet-visible Spectrophotometry Aciclovir Tablets
<2.24>.
アシクロビル錠
＝ MS/MT × AT/AS × 1/C × 1800
Aciclovir Tablets contain not less than 95.0z and
not more than 105.0z of the labeled amount of
MS: Amount (mg) of Aciclovir RS taken, calculated on aciclovir (C8H11N5O3: 225.20).
the anhydrous basis
Method of preparation Prepare as directed under Tablets,
MT: Amount (g) of Aciclovir for Syrup taken
with Aciclovir.
C: Labeled amount (mg) of aciclovir (C8H11N5O3) in 1 g
Assay Weigh accurately an amount of Aciclovir for Syrup,
previously powdered if necessary, equivalent to about 0.2 g
of aciclovir (C8H11N5O3), add 20 mL of diluted sodium hy-
droxide TS (1 in 10), treat with ultrasonic waves to disinte-
grate, then add water to make exactly 200 mL, and filter this Uniformity of dosage units <6.02> It meets the requirement
solution through a membrane filter with a pore size not of the Mass variation test.
exceeding 0.45 mm. Discard the first 2 mL of the filtrate,
pipet 5 mL of the subsequent filtrate, add exactly 5 mL of
the internal standard solution, then add the mobile phase to
in 30 minutes of Aciclovir Tablets is not less than 80z.
Separately, weigh accurately about 10 mg of Aciclovir RS
Start the test with 1 tablet of Aciclovir Tablets, withdraw
(separately determine the water <2.48> in the same manner
not less than 20 mL of the medium at the specified minute
as Aciclovir), add exactly 10 mL of the internal standard
after starting the test, and filter through a membrane filter
solution, then add the mobile phase to make 100 mL, and
with a pore size not exceeding 0.45 mm. Discard the first 10
use this solution as the standard solution. Perform the test
mL of the filtrate, pipet V mL of the subsequent filtrate, add
with 20 mL each of the sample solution and standard solution
water to make exactly V? mL so that each mL contains about
as directed under Liquid Chromatography <2.01> according
8.9 mg of aciclovir (C8H11N5O3), and use this solution as the
to the following conditions, and calculate the ratios, QT
and QS, of the peak area of aciclovir to that of the internal
of aciclovir RS (separately determine the water <2.48> in the
standard.
same manner as Aciclovir), and dissolve in water to make
Amount (mg) of aciclovir (C8H11N5O3) exactly 100 mL. Pipet 4 mL of this solution, add water to
＝ MS × QT/QS × 20 make exactly 100 mL, and use this solution as the standard
the anhydrous basis
under Ultraviolet-visible Spectrophotometry <2.24>.
＝ MS × AT/AS × V?/V × 1/C × 36
length: 254 nm). MS: Amount (mg) of Acyclovir RS taken, calculated on
Column: A stainless steel column 4.6 mm in inside diame- the anhydrous basis
ter and 25 cm in length, packed with octadecylsilanized silica C: Labeled amount (mg) of aciclovir (C8H11N5O3) in 1
gel for liquid chromatography (5 mm in particle diameter). tablet
409 C.
Aciclovir Tablets, and powder. Weigh accurately a portion
Mobile phase: Dissolve 7.8 g of sodium dihydrogen phos-
of the powder, equivalent to about 0.1 g of aciclovir
phate dihydrate and 0.85 g of sodium 1-octanesulfonate in
(C8H11N5O3), add 60 mL of dilute sodium hydroxide TS, and
900 mL of water, adjust to pH 3.0 with phosphoric acid, add
agitate for 15 minutes with the aid of ultrasonic waves, then
water to make 950 mL, and add 50 mL of acetonitrile.
add dilute sodium hydroxide TS to make exactly 100 mL,
Flow rate: Adjust so that the retention time of aciclovir is
and filter. Discard the first 20 mL of filtrate, pipet 15 mL of
about 5 minutes.
the subsequent filtrate, add 50 mL of water and 5.8 mL of 2
mol/L hydrochloric acid TS, and add water to make exactly
chloric acid TS to make exactly 100 mL, and use this solu-
ditions, aciclovir and the internal standard are eluted in this
JP XVII Official Monographs / Aclarubicin Hydrochloride 375
tion as the sample solution. Separately, weigh accurately ties of absorption at the same wavelengths.
about 25 mg of Aciclovir RS (separately determine the water (2) Determine the infrared absorption spectrum of
<2.48> in the same manner as Aciclovir), and dissolve in Aclarubicin Hydrochloride as directed in the potassium bro-
dilute sodium hydroxide TS to make exactly 25 mL. Pipet 15 mide disk method under Infrared Spectrophotometry <2.25>,
mL of this solution, add 50 mL of water and 5.8 mL of 2 and compare the spectrum with the Reference Spectrum:
mol/L hydrochloric acid TS, add water to make exactly 100 both spectra exhibit similar intensities of absorption at the
mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric same wave numbers.
acid TS to make exactly 100 mL, and use this solution as the (3) A solution of Aclarubicin Hydrochloride in methanol
standard solution. Determine the absorbances, AT and AS, at (1 in 200) responds to the Qualitative Tests <1.09> (2) for
255 nm of the sample solution and standard solution as chloride.
D : －146 – －1629(50 mg calcu-
using 0.1 mol/L hydrochloric acid TS as the blank.
lated on the anhydrous basis, water, 10 mL, 100 mm).
pH <2.54> The pH of a solution obtained by dissolving
＝ MS × AT/AS × 4
0.05 g of Aclarubicin Hydrochloride in 10 mL of water is
MS: Amount (mg) of Aciclovir RS taken, calculated on between 5.5 and 6.5.
the anhydrous basis
Purity (1) Clarity and color of solution—Dissolve 0.10 g
Containers and storage Containers—Well-closed contain- of Aclarubicin Hydrochloride in 10 mL of water: the solu-
ers. tion is clear and yellow to pale orange-yellow.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Aclaru-
bicin Hydrochloride according to Method 2, and perform
Aclarubicin Hydrochloride the test. Prepare the control solution with 2.0 mL of Stand-
ard Lead Solution (not more than 20 ppm).
アクラルビシン塩酸塩 (3) Related substances—Dissolve 10 mg of Aclarubicin
Hydrochloride in 10 mL of the mobile phase, and use this
solution as the sample solution. Perform the test with 20 mL
of the sample solution as directed under Liquid Chromatog-
raphy <2.01> according to the following conditions, and de-
termine each peak area by the automatic integration method.
Calculate the amount of the related substances by the area
percentage method: the amount of aklavinone having the
relative retention time of about 0.6 to aclarubicin is not more
than 0.2z, aclacinomycin L1 having the relative retention
time of about 0.75 to aclarubicin is not more than 0.5z, 1-
deoxypyrromycin having the relative retention time of about
1.7 to aclarubicin is not more than 1.5z and aclacinomycin
S1 having the relative retention time of about 2.3 to aclarubi-
cin is not more than 0.5z, and the total amount of the peaks
other than aclarubicin and the peaks mentioned above is not
more than 1.0z of the peak area of aclarubicin.
C42H53NO15.HCl: 848.33
Methyl (1R,2R,4S )-4-{2,6-dideoxy-4-O-[(2R,6S )-6-
Detector: A visible absorption photometer (wavelength:
methyl-5-oxo-3,4,5,6-tetrahydro-2H-pyran-2-yl]-
436 nm).
a-L-lyxo-hexopyranosyl-(1→4)-2,3,6-trideoxy-3-
dimethylamino-a-L-lyxo-hexopyranosyloxy}-2-ethyl-2,5,7-
ter and 30 cm in length, packed with silica gel for liquid
trihydroxy-6,11-dioxo-1,2,3,4-tetrahydrotetracene-
chromatography (10 mm in particle diameter).
1-carboxylate monohydrochloride
[75443-99-1]
259C.
Mobile phase: A mixture of chloroform, methanol, acetic
Aclarubicin Hydrochloride is the hydrochloride of
acid (100), water and triethylamine (6800:2000:1000:200:1).
an anthracycline substance having antitumor activity
Flow rate: Adjust so that the retention time of aclarubicin
produced by the growth of Streptomyces galilaeus.
is about 5 minutes.
It contains not less than 920 mg (potency) and not
Time span of measurement: As long as about 4 times of
more than 975 mg (potency) per mg, calculated on the
the retention time of aclarubicin, beginning after the solvent
anhydrous basis. The potency of Aclarubicin Hydro-
peak.
chloride is expressed as mass (potency) of aclarubicin
(C42H53NO15: 811.87).
Test for required detectability: To 1 mL of the sample so-
Description Aclarubicin Hydrochloride occurs as a yellow lution, add the mobile phase to make 100 mL, and use this
to pale orange-yellow powder. solution as the solution for system suitability test. Pipet 1
It is very soluble in chloroform and in methanol, freely mL of the solution for system suitability test, and add the
soluble in water, and slightly soluble in ethanol (95). mobile phase to make exactly 10 mL. Confirm that the peak
area of aclarubicin obtained from 20 mL of this solution is
equivalent to 7 to 13z of that obtained from 20 mL of the
solution of Aclarubicin Hydrochloride in diluted methanol
(4 in 5) (3 in 100,000) as directed under Ultraviolet-visible
System performance: Dissolve 5 mg of Aclarubicin Hy-
Spectrophotometry <2.24>, and compare the spectrum with
drochloride in 10 mL of 0.1 mol/L hydrochloric acid TS,
the Reference Spectrum: both spectra exhibit similar intensi-
376 Acrinol Hydrate / Official Monographs JP XVII
and allow to stand for 60 minutes. To 1.0 mL of this solu- Ultraviolet-visible Spectrophotometry <2.24>, and compare
tion add 1.0 mL of 0.2 mol/L sodium hydroxide TS, 1.0 mL the spectrum with the Reference Spectrum: both spectra
of phosphate buffer solution (pH 8.0) and 1.0 mL of chlo- exhibit similar intensities of absorption at the same wave-
roform, shake vigorously, and take the chloroform layer. lengths.
When the procedure is run with 20 mL of the chloroform (2) Determine the infrared absorption spectrum of
under the above operating conditions, aclarubicin and 1- Acrinol Hydrate as directed in the potassium bromide disk
deoxypyrromycin are eluted in this order with the resolution method under Infrared Spectrophotometry <2.25>, and com-
between these peaks being not less than 3.0. pare the spectrum with the Reference Spectrum: both spectra
System repeatability: When the test is repeated 5 times exhibit similar intensities of absorption at the same wave
with 20 mL of the sample solution under the above operating numbers.
conditions, the relative standard deviation of the peak area (3) To 5 mL of a solution of Acrinol Hydrate (1 in 100)
of aclarubicin is not more than 2.0z. add 5 mL of dilute sulfuric acid, shake well, allow to stand
for about 10 minutes at room temperature, and filter: the fil-
Water <2.48> Not more than 3.5z (0.1 g, volumetric titra-
trate responds to the Qualitative Tests <1.09> for lactate.
Purity (1) Chloride <1.03>—Dissolve 1.0 g of Acrinol Hy-
drate in 80 mL of water by warming on a water bath, cool,
Assay Weigh accurately an amount of Aclarubicin Hydro- and add 10 mL of sodium hydroxide TS and water to make
chloride, equivalent to about 20 mg (potency), and dissolve 100 mL. Shake well, allow to stand for 30 minutes, filter, to
in diluted methanol (4 in 5) to make exactly 100 mL. Pipet 15 40 mL of the filtrate add 7 mL of dilute nitric acid and water
mL of this solution, add diluted methanol (4 in 5) to make to make 50 mL, and perform the test using this solution as
exactly 100 mL, and use this solution as the sample solution. the test solution. Prepare 50 mL of the control solution with
Separately, weigh accurately an amount of Aclarubicin RS, 4 mL of sodium hydroxide TS, 7 mL of dilute nitric acid,
equivalent to about 20 mg (potency), add 0.6 mL of diluted 0.30 mL of 0.01 mol/L hydrochloric acid VS and sufficient
hydrochloric acid (1 in 250) and diluted methanol (4 in 5) to water (not more than 0.026z).
make exactly 100 mL. Pipet 15 mL of this solution, add (2) Heavy metals <1.07>—Proceed with 1.0 g of Acrinol
diluted methanol (4 in 5) to make exactly 100 mL, and use Hydrate according to Method 2, and perform the test. Pre-
this solution as the standard solution. Perform the test with pare the control solution with 2.0 mL of Standard Lead So-
the sample solution and standard solution as directed under lution (not more than 20 ppm).
Ultraviolet-visible Spectrophotometry <2.24>, and determine (3) Volatile fatty acids—Dissolve 0.5 g of Acrinol Hy-
the absorbances, AT and AS, at 433 nm. drate in a mixture of 20 mL of water and 5 mL of dilute sul-
furic acid, shake well, filter, and heat the filtrate: no odor of
Amount [ mg (potency)] of aclarubicin (C42H53NO15)
volatile fatty acids is perceptible.
＝ MS × AT/AS × 1000
(4) Related substances—Dissolve 10 mg of Acrinol Hy-
MS: Amount [mg (potency)] of Aclarubicin RS taken drate in 25 mL of the mobile phase, and use this solution as
the mobile phase to make exactly 100 mL, and use this solu-
Storage—Light-resistant and at 59C or below.
tion as the standard solution (1). Pipet 1 mL of the standard
solution (1), add the mobile phase to make exactly 10 mL,
and use this solution as the standard solution (2). Perform
Acrinol Hydrate the test with exactly 10 mL each of the sample solution and
standard solutions (1) and (2) as directed under Liquid Chro-
Ethacridine Lactate matography <2.01> according to the following conditions,
and determine each peak area by the automatic integration
アクリノール水和物
method: the area of the peak other than acrinol obtained
with the sample solution is not larger than 3 times the peak
area of acrinol obtained with the standard solution (2), and
the total area of the peaks other than acrinol is not larger
than the peak area of acrinol with the standard solution (1).
C15H15N3O.C3H6O3.H2O: 361.39 Detector: An ultraviolet absorption photometer (wave-
2-Ethoxy-6,9-diaminoacridine monolactate monohydrate length: 268 nm).
[6402-23-9] Column: A stainless steel column 4.6 mm in inside diame-
Acrinol Hydrate contains not less than 98.5z and gel for liquid chromatography (5 mm in particle diameter).
not more than 101.0z of acrinol (C15H15N3O.C3H6O3: Column temperature: A constant temperature of about
343.38), calculated on the anhydrous basis. 259C.
Description Acrinol Hydrate occurs as a yellow crystalline
phate in 900 mL of water, adjust to pH 2.8 with phosphoric
powder.
acid, and add water to make 1000 mL. To 700 mL of this
It is sparingly soluble in water, in methanol and in ethanol
solution add 300 mL of acetonitrile for liquid chromatogra-
(99.5).
phy, and add 1.0 g of sodium 1-octanesulfonate to dissolve.
Melting point: about 2459C (with decomposition).
Flow rate: Adjust so that the retention time of acrinol is
The pH of a solution of Acrinol Hydrate (1 in 100) is
about 15 minutes.
between 5.5 and 7.0.
Time span of measurement: About 3 times as long as the
Identification (1) Determine the absorption spectrum of a retention time of acrinol, beginning after the solvent peak.
solution of Acrinol Hydrate (3 in 250,000) as directed under
JP XVII Official Monographs / Compound Acrinol and Zinc Oxide Oil 377
System suitability— hydrochloric acid, filter after thorough shaking, and to the
Test for required detectability: Confirm that the peak area filtrate add 2 to 3 drops of potassium hexacyanoferrate (II)
of acrinol obtained with 10 mL of the standard solution (2) TS: a white precipitate is formed (zinc oxide).
is equivalent to 7 to 13z of that obtained with 10 mL of the (3) Shake well 0.2 g of Acrinol and Zinc Oxide Oil with
standard solution (1). 20 mL of ethanol (95) and 1 mL of acetic acid (100), centri-
System performance: When the procedure is run with 10 fuge, filter, and use the filtrate as the sample solution. Sepa-
mL of the standard solution (1) under the above operating rately, dissolve 5 mg of acrinol in 50 mL of ethanol (95) and
conditions, the number of theoretical plates and the symme- 2.5 mL of acetic acid (100), and use this solution as the
try factor of the peak of acrinol are not less than 5000 and standard solution. Perform the test with these solutions as
not more than 2.0, respectively. directed under Thin-layer Chromatography <2.03>. Spot 5
System repeatability: When the test is repeated 6 times mL each of the sample solution and standard solution on a
with 10 mL of the standard solution (1) under the above plate of silica gel for thin-layer chromatography. Develop
operating conditions, the relative standard deviation of the the plate with a mixture of 2-propanol and acetic acid (100)
peak area of acrinol is not more than 1.5z. (9:1) to a distance of about 10 cm, and air-dry the plate. Ex-
amine under ultraviolet light (main wavelength: 365 nm): the
Water <2.48> 4.5 – 5.5z (0.2 g, volumetric titration, direct
spots from the sample solution and standard solution exhibit
titration)
a blue fluorescence and show the same R f value.
Assay Transfer about 0.8 g of well-mixed Acrinol and Zinc
Assay Weigh accurately about 0.27 g of Acrinol Hydrate, Oxide Oil, accurately weighed, to a crucible, heat, gradually
dissolve in 5 mL of formic acid, add 60 mL of a mixture of raising the temperature until the mass is throughly charred,
acetic anhydride and acetic acid (100) (1:1), and titrate <2.50> then strongly heat until the residue becomes yellow. After
immediately with 0.1 mol/L perchloric acid VS (potentio- cooling, dissolve the residue by addition of 1 mL of water
metric titration). Perform a blank determination in the same and 1.5 mL of hydrochloric acid, and add water to make
manner, and make any necessary correction. exactly 100 mL. Pipet 20 mL of this solution, add 80 mL of
water, then add sodium hydroxide solution (1 in 50) until
slightly precipitates appear, and add 5 mL of ammonia-
＝ 34.34 mg of C15H15N3O.C3H6O3
ammonium chloride buffer solution (pH 10.7). Titrate <2.50>
Containers and storage Containers—Tight containers. with 0.05 mol/L disodium dihydrogen ethylenediamine
Storage—Light-resistant. tetraacetate VS (indicator: 40 mg of eriochrome black T-
sodium chloride indicator).
Acrinol and Zinc Oxide Oil ethylenediamine tetraacetate VS
＝ 4.069 mg of ZnO
アクリノール・チンク油
Storage—Light-resistant.
Acrinol and Zinc Oxide Oil contains not less than
44.6z and not more than 54.4z of zinc oxide (ZnO:
81.38).
Compound Acrinol and Zinc Oxide
Method of preparation
Oil
Acrinol Hydrate, very finely powdered 10 g
Zinc Oxide Oil 990 g 複方アクリノール・チンク油
To make 1000 g
Prepare by mixing the above ingredients. Acrinol Hydrate
may be mixed after being dissolved in a little amount of Acrinol Hydrate, very finely powdered 10 g
warmed Purified Water or Purified Water in Containers. Zinc Oxide Oil 650 g
Instead of Zinc Oxide Oil adequate amounts of Zinc Oxide Ethyl Aminobenzoate, finely powdered 50 g
and vegetable oil may be used, and an adequate amount of White Beeswax 20 g
Castor Oil or polysorbate 20 may be substituted for a part of Hydrophilic Petrolatum 270 g
the vegetable oil. To make 1000 g
Description Acrinol and Zinc Oxide Oil is a yellowish Prepare by mixing the above ingredients.
white, slimy substance. Separation of a part of its ingredi-
Description Compound Acrinol and Zinc Oxide Oil is light
ents occurs on prolonged standing.
yellow to yellow in color.
Identification (1) Shake well 1 g of Acrinol and Zinc
Identification (1) Shake well 1 g of Compound Acrinol
Oxide Oil with 10 mL of diethyl ether, 2 mL of acetic acid
and Zinc Oxide Oil with 10 mL of diethyl ether, 2 mL of
(100) and 10 mL of water, and separate the water layer.
acetic acid (100) and 10 mL of water, and separate the water
Shake the layer with 5 mL of hydrochloric acid and 2 to 3
layer. Shake the layer with 5 mL of hydrochloric acid and 2
drops of sodium nitrite TS, and allow to stand: a dark red
to 3 drops of sodium nitrite TS, and allow to stand: a dark
color is produced (acrinol).
red color is produced (acrinol).
(2) Place 1 g of Acrinol and Zinc Oxide Oil in a crucible,
(2) Place 1 g of Compound Acrinol and Zinc Oxide Oil
melt by warming, heat, gradually raising the temperature
in a crucible, melt by warming, heat, gradually raising the
until the mass is thoroughly charred, and then ignite
temperature until the mass is thoroughly charred, and then
strongly: a yellow color is produced, and disappears on cool-
ignite strongly: a yellow color is produced, and disappears
ing. To the residue add 10 mL of water and 5 mL of dilute
378 Acrinol and Zinc Oxide Ointment / Official Monographs JP XVII
on cooling. To the residue add 10 mL of water and 5 mL of Containers and storage Containers—Tight containers.
dilute hydrochloric acid, shake well, and filter. To the fil- Storage—Light-resistant.
trate add 2 to 3 drops of potassium hexacyanoferrate (II) TS:
a white precipitate is produced (zinc oxide).
(3) Shake well 0.2 g of Compound Acrinol and Zinc Actinomycin D
Oxide Oil with 20 mL of ethanol (95) and 1 mL of acetic acid
(100), centrifuge, filter, and use the filtrate as the sample アクチノマイシン D
solution. Separately, dissolve 5 mg of acrinol and 25 mg of
ethyl aminobenzoate in 50 mL of ethanol (95) and in 2.5 mL
of acetic acid (100), respectively, and use both solutions as
the standard solutions (1) and (2). Perform the test with
these solutions as directed under Thin-layer Chromatogra-
phy <2.03>. Spot 5 mL each of the sample solution and stand-
ard solutions on a plate of silica gel with fluorescent indica-
tor for thin-layer chromatography. Develop the plate with a
mixture of 2-propanol and acetic acid (100) (9:1) to a dis-
tance of about 10 cm, and air-dry the plate. Examine under
C62H86N12O16: 1255.42
ultraviolet light (main wavelength: 365 nm): the spots from
[50-76-0]
the sample solution and standard solution (1) exhibit a blue
fluorescence, and show the same R f value. Also examine
Actinomycin D is a peptide substance having an-
titumor activity produced by the growth of Strep-
from the sample solution and standard solution (2) exhibit a
tomyces parvulus.
purple color, and show the same R f value.
It, when dried, contains not less than 950 mg (po-
Containers and storage Containers—Tight containers. tency) and not more than 1030 mg (potency) per mg.
Storage—Light-resistant. The potency of Actinomycin D is expressed as mass
(potency) of actinomycin D (C62H86N12O16).
Description Actinomycin D occurs as an orange-red to red
Acrinol and Zinc Oxide Ointment crystalline powder.
It is freely soluble in acetone, sparingly soluble in aceto-
アクリノール・亜鉛華軟膏
nitrile and in methanol, slightly soluble in ethanol (99.5),
and very slightly soluble in water.
Acrinol Hydrate, very finely powdered 10 g
solution of Actinomycin D in methanol (3 in 100,000) as
Zinc Oxide Ointment 990 g
To make 1000 g and compare the spectrum with the Reference Spectrum or
Prepare as directed under Ointments, with the above the spectrum of a solution of Actinomycin D RS prepared in
ingredients. the same manner as the sample solution: both spectra exhibit
Description Acrinol and Zinc oxide Ointment is yellow in (2) Dissolve 0.1 g each of Actinomycin D and Actinomy-
color. cin D RS in 10 mL of acetone, and use these solutions as the
Identification (1) Shake 0.5 g of Acrinol and Zinc Oxide sample solution and standard solution. Perform the test with
Ointment with 5 mL of diethyl ether, 5 mL of dilute hydro- these solutions as directed under Thin-layer Chromatogra-
chloric acid and 2 to 3 drops of sodium nitrite TS, and allow phy <2.03>. Spot 10 mL each of the sample solution and
to stand: a dark red color develops in the water layer standard solution on a plate of silica gel with fluorescent
(acrinol). indicator for thin-layer chromatography. Develop the plate
(2) Ignite 0.5 g of Acrinol and Zinc Oxide Ointment to with a mixture of 1-butanol, water and methanol (4:2:1) to
char, and dissolve the residue in 5 mL of dilute hydrochloric a distance of about 10 cm, and air-dry the plate. Examine
acid: the solution responds to the Qualitative Tests <1.09> for under ultraviolet light (main wavelength: 254 nm): the R f
zinc salt. value of the principal spot from the sample solution is the
(3) Shake 0.5 g of Acrinol and Zinc Oxide Ointment with same as that from the standard solution.
5 mL of diethyl ether, 1 mL of acetic acid (100) and 5 mL of Optical rotation <2.49> [a]20
D : －293 – －3299(after drying,
water, separate the water layer, and use the water layer as 10 mg, methanol, 10 mL, 100 mm).
the sample solution. Dissolve 5 mg of acrinol in 1 mL of
acetic acid (100) and 5 mL of water, and use this solution as Loss on drying <2.41> Not more than 5.0z (1 g, in vacu-
the standard solution. Perform the test with these solutions um, 609C, 3 hours).
as directed under Thin-layer Chromatography <2.03>. Spot 5 Assay Weigh accurately an amount of Actinomycin D and
mL each of the sample solution and standard solution on a Actinomycin D RS, previously dried, equivalent to about 60
plate of silica gel for thin-layer chromatography. Develop mg (potency), dissolve each in the mobile phase to make ex-
the plate with a mixture of diethyl ether, ethanol (95) and actly 50 mL, and use these solutions as the sample solution
acetic acid (100) (40:10:1) to a distance of about 10 cm, and and standard solution. Perform the test with exactly 25 mL
air-dry the plate. Examine under ultraviolet light (main each of the sample solution and standard solution as directed
wavelength: 365 nm): the spots from the sample solution and under Liquid Chromatography <2.01> according to the fol-
the standard solution exhibit a blue fluorescence and show lowing conditions, and determine the peak area of actino-
the same R f value. mycin D, AT and AS, in each solution.
JP XVII Official Monographs / Adrenaline Injection 379
Amount [ mg (potency)] of actinomycin D (C62H86N12O16) exhibit similar intensities of absorption at the same wave
＝ MS × AT/AS × 1000 numbers.
MS: Amount [mg (potency)] of Actinomycin D RS taken Optical rotation <2.49> [a]20
D : －50.0 – －53.59(after dry-
ing, 1 g, 1 mol/L hydrochloric acid TS, 25 mL, 100 mm).
Detector: An ultraviolet absorption photometer (wave- Purity (1) Clarity and color of solution—Dissolve 0.10 g
length: 254 nm). of Adrenaline in 10 mL of dilute hydrochloric acid: the solu-
Column: A stainless steel column 3.9 mm in inside diame- tion is clear, and is not more colored than Matching Fluid A.
ter and 30 cm in length, packed with octadecylsilanized silica (2) Heavy metals <1.07>—Proceed with 1.0 g of Adrena-
gel for liquid chromatography (10 mm in particle diameter). line according to Method 4, and perform the test. Prepare
Column temperature: A constant temperature of about the control solution with 2.0 mL of Standard Lead Solution
259 C. (not more than 20 ppm).
Mobile phase: A mixture of 0.02 mol/L acetic acid-sodi- (3) Adrenalone—Dissolve 50 mg of Adrenaline in 0.05
um acetate TS and acetonitrile (25:23). mol/L hydrochloric acid TS to make exactly 25 mL, and de-
Flow rate: Adjust so that the retention time of actinomy- termine the absorbance of this solution at 310 nm as directed
cin D is about 23 minutes. under Ultraviolet-visible Spectrophotometry <2.24>: it is not
System suitability— more than 0.2.
System performance: When the procedure is run with 25 (4) Noradrenaline—Dissolve 0.20 g of Adrenaline in 1
mL of the standard solution under the above operating con- mL of formic acid, add methanol to make exactly 10 mL,
ditions, the number of theoretical plates and the symmetry and use this solution as the sample solution. Separately, dis-
factor of the peak of actinomycin D are not less than 2000 solve 8.0 mg of Noradrenaline Bitartrate RS in methanol to
and not more than 1.5, respectively. make exactly 10 mL, and use this solution as the standard
System repeatability: When the test is repeated 5 times solution. Perform the test with these solutions as directed
with 25 mL of the standard solution under the above operat- under Thin-layer Chromatography <2.03>. Spot 5 mL each of
ing conditions, the relative standard deviation of the peak the sample solution and standard solution on a plate of silica
area of actinomycin D is not more than 2.0z. gel with fluorescent indicator for thin-layer chromatogra-
phy. Develop the plate with a mixture of 1-butanol, water
and formic acid (7:2:1) to a distance of about 10 cm, and air-
dry the plate. Spray evenly Folin's TS on the plate: the spot
obtained from the sample solution, corresponding to the
spot obtained from the standard solution, is not more in-
Adrenaline tense than the spot from the standard solution.
Epinephrine Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-
um, silica gel, 18 hours).
アドレナリン
Assay Weigh accurately about 0.3 g of Adrenaline, previ-
ously dried, dissolve in 50 mL of acetic acid (100), and titrate
<2.50> with 0.1 mol/L perchloric acid VS (potentiometric
titration). Perform a blank determination in the same
manner, and make any necessary correction.
C9H13NO3: 183.20
4-[(1R)-1-Hydroxy-2-(methylamino)ethyl]benzene-1,2-diol
＝ 18.32 mg of C9H13NO3
[51-43-4]
Adrenaline, when dried, contains not less than Storage—Light-resistant, and under Nitrogen atmosphere.
98.0z and not more than 101.0z of adrenaline
(C9H13NO3).
Description Adrenaline occurs as a white to grayish white Adrenaline Injection
crystalline powder.
It is freely soluble in formic acid and in acetic acid (100),
Epinephrine Injection
very slightly soluble in water, and practically insoluble in
アドレナリン注射液
methanol and in ethanol (99.5).
It dissolves in dilute hydrochloric acid.
It gradually changes to brown in color by air and by light. Adrenaline Injection is an aqueous injection.
It contains not less than 0.085 w/vz and not more
than 0.115 w/vz of adrenaline (C9H13NO3: 183.20).
solution of Adrenaline in 0.01 mol/L hydrochloric acid TS
(1 in 25,000) as directed under Ultraviolet-visible Spectro- Method of preparation Dissolve Adrenaline in diluted Hy-
photometry <2.24>, and compare the spectrum with the Ref- drochloric Acid (9 in 10,000), and prepare as directed under
erence Spectrum: both spectra exhibit similar intensities of Injections.
absorption at the same wavelengths.
Description Adrenaline Injection is a colorless, clear liq-
uid.
Adrenaline as directed in the potassium bromide disk
It changes gradually to pale red and then to brown on
method under Infrared Spectrophotometry <2.25>, and com-
exposure to air and light.
pare the spectrum with the Reference Spectrum: both spectra
pH: 2.3 – 5.0
380 Adrenaline Solution / Official Monographs JP XVII
Identification (1) To 1 mL of Adrenaline Injection add exposure to air and light.
4 mL of water and 1 drop of iron (III) chloride TS: a deep pH: 2.3 – 5.0
green color is produced, and it gradually changes to red.
Identification Proceed as directed in the Identification
(2) Place 1 mL each of Adrenaline Injection in test tubes
under Adrenaline Injection.
A and B, and proceed as directed in the Identification (2)
under Adrenaline. Assay Proceed as directed in the Assay under Adrenaline
Injection.
Amount (mg) of adrenaline (C9H13NO3)
Assay Pipet 30 mL of Adrenaline Injection into a separa-
＝ M × {0.5 ＋ (0.5 × |[a]20D |)/93} × 0.592
tor, add 25 mL of carbon tetrachloride, shake vigorously for
1 minute, allow the liquids to separate, and discard the car- Containers and storage Containers—Tight containers.
bon tetrachloride. Repeat this procedure three times. Rinse Storage—Light-resistant.
the stopper and mouth of the separator with a small amount
of water. Add 0.2 mL of starch TS, then while swirling the
separator add iodine TS dropwise until a persistent blue Afloqualone
color develops, and immediately add sodium thiosulfate TS
to discharge the blue color. Add 2.1 g of sodium hydrogen アフロクアロン
carbonate to the liquid in the separator, preventing it from
coming in contact with the mouth of the separator, and
shake until most of the sodium hydrogen carbonate dis-
solves. Rapidly inject 1.0 mL of acetic anhydride into the
contents of the separator. Immediately stopper the separator
loosely, and allow to stand until the evolution of gas ceases.
Shake vigorously, allow to stand for 5 minutes, extract with
C16H14FN3O: 283.30
six 25-mL portions of chloroform, and filter each chlo-
6-Amino-2-fluoromethyl-3-(2-tolyl)-3H-quinazolin-4-one
roform extract through a pledget of absorbent cotton.
[56287-74-2]
Evaporate the combined chloroform extracts on a water bath
in a current of air to 3 mL, completely transfer this residue
Afloqualone, when dried, contains not less than
by means of small portions of chloroform to a tared beaker,
98.5z of afloqualone (C16H14FN3O).
and heat again to evaporate to dryness. Dry the residue at
1059C for 30 minutes, cool in a desiccator (silica gel), and Description Afloqualone occurs as white to light yellow,
accurately measure the mass M (mg) of the dried residue. crystals or crystalline powder.
Dissolve in chloroform to make exactly 5 mL, and determine It is soluble in acetonitrile, sparingly soluble in ethanol
the optical rotation <2.49> [a]20
D using a 100-mm cell. (99.5), and practically insoluble in water.
It is gradually colored by light.
Amount (mg) of adrenaline (C9H13NO3)
Melting point: about 1979 C (with decomposition).
＝ M × {0.5 ＋ (0.5 × |[a]20D |)/93} × 0.592
Identification (1) Conduct this procedure without ex-
Containers and storage Containers—Hermetic containers,
posure to light, using light-resistant containers. Determine
and colored containers may be used.
the absorption spectrum of a solution of Afloqualone in
ethanol (99.5) (1 in 150,000) as directed under Ultraviolet-
Adrenaline Solution tensities of absorption at the same wavelengths.
Epinephrine Solution Afloqualone, previously dried, as directed in the potassium
bromide disk method under Infrared Spectrophotometry
アドレナリン液
<2.25>, and compare the spectrum with the Reference Spec-
trum: both spectra exhibit similar intensities of absorption at
Adrenaline Solution contains not less than 0.085 the same wave numbers.
w/vz and not more than 0.115 w/vz of adrenaline
Purity (1) Acidity or alkalinity—Take 1.0 g of Afloqua-
(C9H13NO3: 183.20)
lone in a light-resistant vessel, add 20 mL of freshly boiled
Method of preparation and cooled water, shake well, and filter. To 10 mL of the fil-
trate add 2 drops of bromothymol blue TS: a yellow color
Adrenaline 1g
develops. The color changes to blue by adding 0.20 mL of
Sodium Chloride 8.5 g
0.01 mol/L sodium hydroxide TS.
Diluted Hydrochloric Acid (9 in 100) 10 mL
(2) Heavy metals <1.07>—Proceed with 2.0 g of Afloqua-
Stabilizer a suitable quantity
lone in a platinum crucible according to Method 2, and per-
Preservative a suitable quantity
form the test. Prepare the control solution with 2.0 mL of
Purified Water or Purified
Standard Lead Solution (not more than 10 ppm).
Water in Containers a sufficient quantity
(3) Related substances—Conduct this procedure without
To make 1000 mL exposure to light, using light-resistant vessels. Dissolve 10
Prepare by mixing the above ingredients. mg of Afloqualone in 25 mL of the mobile phase, and use
this solution as the sample solution. Pipet 3 mL of the sam-
Description Adrenaline Solution is clear, colorless or ple solution, add the mobile phase to make exactly 100 mL.
slightly reddish liquid. Pipet 2 mL of this solution, add the mobile phase to make
It changes gradually to pale red and then to brown on exactly 20 mL, and use this solution as the standard solution.
JP XVII Official Monographs / Ajmaline 381
Perform the test with exactly 20 mL each of the sample solu-

tion and standard solution as directed under Liquid Chroma- Ajmaline
tography <2.01> according to the following conditions, and
calculate the areas of each peak by the automatic integration アジマリン
method: the total of the peak areas other than afloqualone
from the sample solution is not larger than the peak area of
afloqualone from the standard solution.
length: 254 nm).
ter and 15 cm in length, packed with octylsilanized silica gel
for liquid chromatography (5 mm in particle diameter).
C20H26N2O2: 326.43
(17R,21R)-Ajmalan-17,21-diol
409 C.
[4360-12-7]
Mobile phase: Dissolve 7.2 g of disodium hydrogen
phosphate dodecahydrate in 1000 mL of water, adjust to pH
Ajmaline, when dried, contains not less than 96.0z
5.5 with diluted phosphoric acid (1 in 10). To 600 mL of this
of ajmaline (C20H26N2O2).
solution add 400 mL of acetonitrile.
Flow rate: Adjust so that the retention time of afloqua- Description Ajmaline occurs as a white to pale yellow
lone is about 5.5 minutes. crystalline powder. It is odorless, and has a bitter taste.
Time span of measurement: About 4 times as long as the It is freely soluble in acetic anhydride and in chloroform,
retention time of afloqualone, beginning after the solvent sparingly soluble in methanol, in ethanol (95), in acetone
peak. and in diethyl ether, and very slightly soluble in water.
System suitability— It dissolves in dilute hydrochloric acid.
Test for required detectability: Pipet 5 mL of the standard Melting point: about 1959C (with decomposition).
solution, add the mobile phase to make exactly 25 mL, and
Identification (1) Dissolve 0.05 g of Ajmaline in 5 mL of
confirm that the peak area of afloqualone obtained from
methanol, and use this solution as the sample solution. Add
20 mL of this solution is equivalent to 15 to 25z of that of
3 mL of nitric acid to 1 mL of the sample solution: a deep
afloqualone obtained from 20 mL of the standard solution.
red color develops.
System performance: Dissolve 0.01 g of Afloqualone in a
(2) Spot the sample solution of (1) on filter paper, and
suitable amount of the mobile phase, add 5 mL of a solution
spray Dragendorff's TS: an orange color develops.
of propyl parahydroxybenzoate in the mobile phase (1 in
2000) and the mobile phase to make 100 mL. When the Absorbance <2.24> E 11zcm (249 nm): 257 – 271 (after drying,
procedure is run with 20 mL of this solution under the above 2 mg, ethanol (95), 100 mL).
operating conditions, afloqualone and propyl parahydroxy- E 11zcm (292 nm): 85 – 95 (after drying, 2 mg, ethanol (95),
benzoate are eluted in this order with the resolution between 100 mL).
these peaks being not less than 4.
D : ＋136 – ＋1519(after drying,
0.5 g, chloroform, 50 mL, 100 mm).
ing conditions, the relative standard deviation of the peak Purity Related substances—Dissolve 0.10 g of Ajmaline in
areas of afloqualone is not more than 5z. 10 mL of chloroform, and use this solution as the sample so-
lution. Pipet 1 mL of the sample solution, add chloroform to
Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-
um, 609C, 2 hours).
solution. Perform the test with these solutions as directed
Residue on ignition <2.44> Not more than 0.1z (1.0 g, under Thin-layer Chromatography <2.03>. Spot 10 mL each
platinum crucible). of the sample solution and standard solution on a plate of
silica gel with fluorescent indicator for thin-layer chromatog-
Assay Weigh accurately about 0.4 g of Afloqualone, previ-
raphy. Develop the plate with a mixture of chloroform, ace-
ously dried, dissolve in 10 mL of hydrochloric acid and 40
tone and diethylamine (5:4:1) to a distance of about 10 cm,
mL of water, and add 10 mL of a solution of potassium bro-
mide (3 in 10). After cooling at 159C or below, titrate <2.50>
with 0.1 mol/L sodium nitrite VS according to the potentio-
from the sample solution are not more intense than the spot
metric titration or amperometric titration under the Electro-
from the standard solution.
metric Titration method.
Loss on drying <2.41> Not more than 1.0z (0.6 g, in vacu-
Each mL of 0.1 mol/L sodium nitrite
um, 809C, 3 hours).
＝ 28.33 mg of C16H14FN3O
Storage—Light-resistant. Assay Weigh accurately about 0.3 g of Ajmaline, previ-
ously dried, dissolve in 50 mL of acetic anhydride and 50 mL
of acetone for nonaqueous titration, and titrate <2.50> with
0.05 mol/L perchloric acid VS (potentiometric titration).
Perform a blank determination, and make any necessary
correction.
＝ 16.32 mg of C20H26N2O2
382 Ajmaline Tablets / Official Monographs JP XVII
Containers and storage Containers—Well-closed contain- standard solution. Determine the absorbances, AT and AS,
ers. of the sample solution and standard solution at 288 nm as di-
Storage—Light-resistant. rected under Ultraviolet-visible Spectrophotometry <2.24>.
of ajmaline (C20H26N2O2)
Ajmaline Tablets ＝ MS × AT/AS × V?/V × 1/C × 180
アジマリン錠 MS: Amount (mg) of ajmaline for assay taken
C: Labeled amount (mg) of ajmaline (C20H26N2O2) in 1
tablet
Ajmaline Tablets contain not less than 90.0z and
not more than 110.0z of the labeled amount of Assay Weigh accurately and powder not less than 20
ajmaline (C20H26N2O2: 326.43). Ajmaline Tablets. Weigh accurately a portion of the powder,
equivalent to about 0.3 g of ajmaline (C20H26N2O2), add 15
mL of ammonia solution (28), and extract with four 25-mL
with Ajmaline.
portions of chloroform. Combine the chloroform extracts,
Identification (1) Shake a quantity of powdered Ajmaline wash with 10 mL of water, add 5 g of anhydrous sodium
Tablets, equivalent to 0.1 g of Ajmaline, with 30 mL of chlo- sulfate, shake well, and filter. Wash the container and the
roform, and filter. Evaporate the filtrate on a water bath to residue with two 10-mL portions of chloroform, and filter.
dryness. With the residue, proceed as directed in the Identifi- Evaporate the combined filtrate on a water bath to dryness,
cation under Ajmaline. dissolve the residue in 50 mL of acetic anhydride and 50 mL
(2) Dissolve 0.01 g of the residue of (1) in 100 mL of of acetone for nonaqueous titration, and titrate <2.50> with
ethanol (95). To 10 mL of this solution add ethanol (95) to 0.05 mol/L perchloric acid VS (potentiometric titration).
make 50 mL, and determine the absorption spectrum of the Perform a blank determination, and make any necessary
solution as directed under Ultraviolet-visible Spectropho- correction.
tometry <2.24>: it exhibits maxima between 247 nm and
251 nm and between 291 nm and 294 nm, and a minimum
＝ 16.32 mg of C20H26N2O2
between 269 nm and 273 nm.
ers.
To 1 tablet of Ajmaline Tablets add 150 mL of 2nd fluid
for dissolution test, shake to disintegrate the tablet, then add
2nd fluid for dissolution test to make exactly 200 mL, and Alacepril
filter this solution through a membrane filter with a pore size
アラセプリル
not exceeding 0.8 mm. Discard the first 10 mL of the filtrate,
pipet V mL of the subsequent filtrate equivalent to about 0.5
mg of ajmaline (C20H26N2O2), add 2nd fluid for dissolution
test to make exactly 10 mL, and use this solution as the sam-
ple solution. Separately, weigh accurately about 25 mg of
ajmaline for assay, previously dried in vacuum at 809C for
3 hours, dissolve in 2nd fluid for dissolution test to make ex-
actly 500 mL, and use this solution as the standard solution.
C20H26N2O5S: 406.50
Determine the absorbances at 288 nm, AT and AS, of the
(2S )-2-{(2S )-1-[(2S )-3-(Acetylsulfanyl)-
sample solution and standard solution as directed under
2-methylpropanoyl]pyrrolidine-2-carbonyl}amino-
Ultraviolet-visible Spectrophotometry <2.24>.
3-phenylpropanoic acid
Amount (mg) of ajmaline (C20H26N2O2) [74258-86-9]
＝ MS × AT/AS × 1/V × 4
Alacepril, when dried, contains not less than 98.5z
MS: Amount (mg) of ajmaline for assay taken
and not more than 101.0z of alacepril (C20H26N2O5S).
Dissolution <6.10> When the test is performed at 100 revo-
Description Alacepril occurs as white, crystals or crystal-
lutions per minute according to the Paddle method, using
line powder.
It is freely soluble in methanol, soluble in ethanol (95),
medium, the dissolution rate in 60 minutes of Ajmaline
and slightly soluble in water.
Tablets is not less than 75z.
Start the test with 1 tablet of Ajmaline Tablets, withdraw
not less than 20 mL of the medium at the specified minute Identification (1) To 20 mg of Alacepril add 0.1 g of so-
after starting the test, and filter through a membrane filter dium hydroxide, and heat gradually to melt: the gas evolved
with a pore size not exceeding 0.8 mm. Discard the first 10 changes the color of a moisten red litmus paper to blue.
mL of the filtrate, pipet V mL of the subsequent filtrate, add After cooling, to the melted substance add 2 mL of water,
the dissolution medium to make exactly V? mL so that each shake, and add 1 mL of lead (II) acetate TS: a brown to
mL contains about 56 mg of ajmaline (C203H26N2O2), and use black precipitate is formed.
this solution as the sample solution. Separately, weigh accu- (2) Determine the infrared absorption spectrum of
rately about 28 mg of ajmaline for assay, previously dried in Alacepril, previously dried, as directed in the potassium bro-
vacuum at 809 C for 3 hours, dissolve in the dissolution me- mide disk method under Infrared Spectrophotometry <2.25>,
dium to make exactly 500 mL, and use this solution as the and compare the spectrum with the Reference Spectrum:
JP XVII Official Monographs / Alacepril Tablets 383
both spectra exhibit similar intensities of absorption at the with the resolution between these peaks being not less than 7.
same wave numbers. System repeatability: When the test is repeated 6 times
Optical rotation <2.49> [a]20D : －81 – －859 (after drying,
0.25 g, ethanol (95), 25 mL, 100 mm).
area of alacepril is not more than 2.0z.
Purity (1) Chloride <1.03>—Dissolve 0.5 g of Alacepril in 3 hours).
30 mL of methanol, add 6 mL of dilute nitric acid and water
to make 50 mL, and perform the test with this solution as the
test solution. Prepare the control solution as follows: to 0.30 Assay Weigh accurately about 0.6 g of Alacepril, previ-
mL of 0.01 mol/L hydrochloric acid VS add 30 mL of meth- ously dried, dissolve in 75 mL of a mixture of methanol and
anol, 6 mL of dilute nitric acid and water to make 50 mL water (2:1), and titrate <2.50> with 0.1 mol/L sodium hy-
(not more than 0.021z). droxide VS (potentiometric titration). Perform a blank de-
(2) Sulfate <1.14>—Dissolve 0.5 g of Alacepril in 30 mL termination in the same manner, and make any necessary
of methanol, add 1 mL of dilute hydrochloric acid and water correction.
to make 50 mL, and perform the test using this solution as
the test solution. Prepare the control solution as follows: to
＝ 40.65 mg of C20H26N2O5S
0.50 mL of 0.005 mol/L sulfuric acid VS add 30 mL of
methanol, 1 mL of dilute hydrochloric acid and water to Containers and storage Containers—Tight containers.
make 50 mL (not more than 0.048z).
(3) Heavy metals <1.07>—Proceed with 1.0 g of Alacepril
according to Method 2, and perform the test. Prepare the Alacepril Tablets
more than 20 ppm). アラセプリル錠
(4) Related substances—Dissolve 50 mg of Alacepril in 5
mL of ethanol (95), and use this solution as the sample solu-
Alacepril Tablets contain not less than 95.0z and
tion. Pipet 1 mL of the sample solution, add ethanol (95) to
alacepril (C20H26N2O5S: 406.50).
solution. Perform the test with exactly 10 mL each of the
sample solution and standard solution as directed under Liq- Method of preparation Prepare as directed under Tablets,
uid Chromatography <2.01> according to the following con- with Alacepril.
ditions, and determine each peak area by the automatic in-
Identification Shake well a quantity of powdered Alacepril
tegration method: the area of the peak other than alacepril
Tablets, equivalent to 0.1 g of Alacepril, with 10 mL of
from the sample solution is not larger than 2/5 times the
ethanol (95), filter, and use the filtrate as the sample solu-
peak area of alacepril from the standard solution, and the
tion. Separately, dissolve 10 mg of alacepril in 1 mL of
total area of the peaks other than alacepril from the sample
ethanol (95), and use this solution as the standard solution.
solution is not larger than the peak area of alacepril from the
standard solution. For the areas of the peaks, having the
relative retention times of about 2.3 and about 2.6 to
solution and standard solution on a plate of silica gel with
alacepril, multiply their relative response factors, 1.5 and
fluorescent indicator for thin-layer chromatography, de-
1.9, respectively.
velop the plate with a mixture of ethanol (99.5) and hexane
(2:1) to a distance of about 10 cm, and air-dry the plate. Ex-
amine under ultraviolet light (main wavelength: 254 nm): the
length: 254 nm).
principal spot from the sample solution and the spot from
the standard solution show the same color tone and R f
value.
Column temperature: A constant temperature of about Uniformity of dosage units <6.02> Perform the Mass varia-
409 C. tion test, or the Content uniformity test according to the fol-
Mobile phase: A mixture of diluted acetic acid (100) (1 in lowing method: it meets the requirement.
100), acetonitrile, methanol and tetrahydrofuran (6:2:1:1). To 1 tablet of Alacepril Tablets add 2 mL of water, dis-
Flow rate: Adjust so that the retention time of alacepril is perse the particle with the aid of ultrasonic wave, and
about 5 minutes. add exactly 2 mL of the internal standard solution for every
Time span of measurement: About 3 times as long as the 10 mg of alacepril (C20H26N2O5S). To this solution add a
retention time of alacepril, beginning after the solvent peak. suitable amount of methanol, extract for 15 minutes with the
System suitability— aid of ultrasonic wave while occasional shaking, and shake
Test for required detectability: To exactly 4 mL of the more 15 minutes. Add methanol to make V mL so that
standard solution add ethanol (95) to make exactly 10 mL. each mL of the solution contains about 0.5 mg of alacepril
Confirm that the peak area of alacepril obtained with 10 mL (C20H26N2O5S), centrifuge, and use the supernatant liquid
of this solution is equivalent to 30 to 50z of that obtained as the sample solution. Separately, weigh accurately about
with 10 mL of the standard solution. 25 mg of alacepril for assay, previously dried at 1059C for
System performance: Dissolve 20 mg of Alacepril in 50 3 hours, add exactly 5 mL of the internal standard solution
mL of a solution of propyl parahydroxybenzoate in ethanol and methanol to make 50 mL, and use this solution as the
(95) (1 in 80,000). When the procedure is run with 10 mL of standard solution. Perform the test with 10 mL each of the
this solution under the above operating conditions, alacepril sample solution and standard solution as directed under
and propyl parahydroxybenzoate are eluted in this order Liquid Chromatography <2.01> according to the following
384 L-Alanine / Official Monographs JP XVII
conditions, and calculate the ratios, QT and QS, of the peak Operating conditions—
area of alacepril to that of the internal standard. Detector: An ultraviolet absorption photometer (wave-
length: 254 nm).
Amount (mg) of alacepril (C20H26N2O5S)
＝ MS × QT/QS × V/50
MS: Amount (mg) of alacepril for assay taken gel for liquid chromatography (5 mm in particle diameter).
Internal standard solution—A solution of propyl parahy-
409C.
droxybenzoate in methanol (3 in 20,000).
Mobile phase: A mixture of diluted acetic acid (100) (1 in
100), acetonitrile, methanol and tetrahydrofuran (13:5:1:1).
Proceed as directed in the operating conditions in the
Flow rate: Adjust so that the retention time of alacepril is
Assay.
about 6 minutes.
Proceed as directed in the system suitability in the Assay.
Dissolution <6.10> When the test is performed at 50 revolu- mL of the standard solution under the above operating con-
tions per minute according to the Paddle method, using 900 ditions, alacepril and the internal standard are eluted in this
mL of water as the dissolution medium, the dissolution rate order with the resolution between these peaks being not less
of a 12.5-mg tablet and a 25-mg tablet in 30 minutes is not than 7.
less than 75z, and that of a 50-mg tablet in 30 minutes is System repeatability: When the test is repeated 6 times
not less than 70z. with 10 mL of the standard solution under the above operat-
Start the test with 1 tablet of Alacepril Tablets, withdraw ing conditions, the relative standard deviation of the ratio of
not less than 20 mL of the medium at the specified minute the peak area of alacepril to that of the internal standard is
after starting the test, and filter through a membrane filter not more than 1.0z.
14 mg of alacepril (C20H26N2O5S), and use this solution as the
sample solution. Separately, weigh accurately about 14 mg L-Alanine
of alacepril for assay, previously dried at 1059C for 3 hours,
L-アラニン
dissolve in 2 mL of methanol, and add water to make exactly
100 mL. Pipet 5 mL of this solution, add water to make ex-
Determine the absorbances, AT1 and AS1, at 230 nm, and AT2
and AS2, at 300 nm, of the sample solution and standard so-
C3H7NO2: 89.09
lution as directed under Ultraviolet-visible Spectrophotome-
(2S )-2-Aminopropanoic acid
try <2.24>, using water as the blank.
[56-41-7]
Dissolution rate (z) with respect to the labeled amount of
alacepril (C20H26N2O5S) L-Alanine, when dried, contains not less than 98.5z
＝ MS × (AT1 － AT2)/(AS1 － AS2) × V?/V × 1/C × 90 and not more than 101.0z of L-alanine (C3H7NO2).
MS: Amount (mg) of alacepril for assay taken Description L-Alanine occurs as white, crystals or crystal-
C: Labeled amount (mg) of alacepril (C20H26N2O5S) in 1 line powder. It has a slightly sweet taste.
tablet It is freely soluble in water and in formic acid, and practi-
cally insoluble in ethanol (99.5).
Assay Weigh accurately, and powder not less than 20
It dissolves in 6 mol/L hydrochloric acid TS.
Alacepril Tablets. Weigh accurately a portion of the powder,
equivalent to about 50 mg of alacepril (C20H26N2O5S), Identification Determine the infrared absorption spectrum
moisten with 2 mL of water, add exactly 3 mL of the internal of L-Alanine as directed in the potassium bromide disk
standard solution and 40 mL of methanol, extract for 15 method under Infrared Spectrophotometry <2.25>, and com-
minutes with the aid of ultrasonic wave, cool, and add meth- pare the spectrum with the Reference Spectrum: both spectra
anol to make 50 mL. Centrifuge this solution, and use the exhibit similar intensities of absorption at the same wave
supernatant liquid as the sample solution. Separately, weigh numbers.
accurately about 50 mg of alacepril for assay, previously
D : ＋13.5 – ＋15.59(after dry-
dried at 1059C for 3 hours, add exactly 3 mL of the internal
ing, 2.5 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm).
standard solution, dissolve with methanol to make 50 mL,
and use this solution as the standard solution. Perform the pH <2.54> Dissolve 1.0 g of L-Alanine in 20 mL of water:
test with 10 mL each of the sample solution and standard so- the pH of the solution is between 5.7 and 6.7.
of L-Alanine in 10 mL of water: the solution is clear and
QT and QS, of the peak area of alacepril to that of the inter-
colorless.
nal standard.
(2) Chloride <1.03>—Perform the test with 0.5 g of
Amount (mg) of alacepril (C20H26N2O5S) ＝ MS × QT/QS L-Alanine. Prepare the control solution with 0.30 mL of
0.01 mol/L hydrochloric acid VS (not more than 0.021z).
MS: Amount (mg) of alacepril for assay taken
(3) Sulfate <1.14>—Perform the test with 0.6 g of
Internal standard solution—A solution of propyl parahy- L-Alanine. Prepare the control solution with 0.35 mL of
droxybenzoate in methanol (1 in 2000). 0.005 mol/L sulfuric acid VS (not more than 0.028z).
JP XVII Official Monographs / L-Alanine 385
L-Alanine. Prepare the control solution with 5.0 mL of

Standard Ammonium Solution (not more than 0.02z). Mobile Mobile Mobile Mobile Mobile
phase A phase B phase C phase D phase E
(5) Heavy metals <1.07>—Proceed with 1.0 g of
L-Alanine according to Method 1, and perform the test. Citric acid 19.80 g 22.00 g 12.80 g 6.10 g —
monohydrate
Solution (not more than 10 ppm). Trisodium citrate 6.19 g 7.74 g 13.31 g 26.67 g —
dihydrate
(6) Iron <1.10>—Prepare the test solution with 1.0 g of
Sodium chlo-
L-Alanine according to Method 1, and perform the test ride 5.66 g 7.07 g 3.74 g 54.35 g —
according to Method A. Prepare the control solution with Sodium hydroxide — — — — 8.00 g
Ethanol (99.5) 130 mL 20 mL 4 mL — 100 mL
(7) Related substances—Weigh accurately about 0.5 g of
Thiodiglycol 5 mL 5 mL 5 mL — —
L-Alanine, dissolve in 0.5 mL of hydrochloric acid and water
Benzyl alcohol — — — 5 mL —
0.02 mol/L hydrochloric acid TS to make exactly 50 mL, Lauromacrogol 4 mL 4 mL 4 mL 4 mL 4 mL
solution (1 in 4)
and use this solution as the sample solution. Separately,
Water Appropriate Appropriate Appropriate Appropriate Appropriate
accurately measure 2.5 mmol amounts of L-aspartic acid, amount amount amount amount amount
L-threonine, L-serine, L-glutamic acid, glycine, L-alanine,
L-cystine, L-valine, L-methionine, L-isoleucine, L-leucine,
Total volume 1000 mL 1000 mL 1000 mL 1000 mL 1000 mL
L-tyrosine, L-phenylalanine, L-lysine hydrochloride, ammo-
nium chloride, L-histidine and L-arginine, dissolve in 0.1 Changing mobile phases: When the procedure is run with
mol/L hydrochloric acid TS to make exactly 1000 mL, and 20 mL of the standard solution under the above operating
use this solution as the standard stock solution. Pipet 5 mL conditions, switchover in sequence to mobile phases A, B, C,
of the standard stock solution, add 0.02 mol/L hydrochloric D and E so that aspartic acid, threonine, serine, glutamic
acid TS to make exactly 100 mL. Pipet 4 mL of this solution, acid, glycine, alanine, cystine, valine, methionine, isoleu-
add 0.02 mol/L hydrochloric acid TS to make exactly 50 cine, leucine, tyrosine, phenylalanine, lysine, ammonia,
mL, and use this solution as the standard solution. Perform histidine, and arginine are eluted in this order with the reso-
the test with exactly 20 mL each of the sample solution and lution between the peaks of isoleucine and leucine being not
standard solution as directed under Liquid Chromatography less than 1.2.
<2.01> according to the following conditions. Based on the Reaction reagents: Dissolve 204 g of lithium acetate dihy-
peak heights obtained from the sample solution and stand- drate in water, add 123 mL of acetic acid (100) and 401 mL
ard solution, determine the mass of the amino acids other of 1-methoxy-2-propanol, and water to make 1000 mL,
than alanine contained in 1 mL of the sample solution, and introduce nitrogen for 10 minutes, and use this solution as
calculate the mass percent: the amount of each amino acid solution (I). Separately, add 39 g of ninhydrin to 979 mL of
other than alanine is not more than 0.1z. 1-methoxy-2-propanol, introduce nitrogen for 5 minutes,
Operating conditions— add 81 mg of sodium borohydride, introduce nitrogen for 30
Detector: A visible spectrophotometer (wavelength: 570 minutes, and use this solution as solution (II). To 1 volume
nm). of solution (I) add 1 volume of solution (II). Prepare before
Column: A stainless steel column 4.6 mm in inside diame- use.
ter and 8 cm in length, packed with strongly acidic ion- Flow rate of mobile phase: 0.20 mL per minute.
exchange resin for liquid chromatography (Na type) com- Flow rate of reaction reagent: 0.24 mL per minute.
posed with a sulfonated polystyrene copolymer (3 mm in System suitability—
particle diameter). System performance: When the procedure is run with
Column temperature: A constant temperature of about 20 mL of the standard solution under the above operating
579 C. conditions, the resolution between the peaks of glycine and
Chemical reaction bath temperature: A constant tempera- alanine is not less than 1.2.
ture of about 1309C. System repeatability: When the test is repeated 6 times
Reaction time: About 1 minute. with 20 mL of the standard solution under the above operat-
Mobile phase: Prepare mobile phases A, B, C, D and E ing conditions, the relative standard deviations of the peak
according to the following table, and add 0.1 mL of capric height and retention time of each amino acid obtained from
acid to each mobile phase. the standard solution are not more than 5.0z and not more
than 1.0z, respectively.
3 hours).
Assay Weigh accurately about 90 mg of L-Alanine, previ-
ously dried, dissolve in 3 mL of formic acid, add 50 mL of
acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric
acid VS (potentiometric titration). Perform a blank determi-
nation in the same manner, and make any necessary correc-
tion.
＝ 8.909 mg of C3H7NO2
386 Albumin Tannate / Official Monographs JP XVII
and not less than 11.1z and not more than 13.0z of
Albumin Tannate aluminum (Al: 26.98).
Description Aldioxa occurs as a white powder.
タンニン酸アルブミン
It is practically insoluble in water and in ethanol (99.5).
Albumin Tannate is a compound of tannic acid and A solution of Aldioxa in sodium fluoride-hydrochloric
a protein. acid TS (1 in 100) shows no optical rotation.
The label states the origin of the protein of Albumin Melting point: about 2309 C (with decomposition).
Tannate.
Description Albumin Tannate occurs as a light brown trum of Aldioxa, previously dried, as directed in the potas-
powder. It is odorless, or has a faint, characteristic odor. sium bromide disk method under Infrared Spectrophotome-
It is practically insoluble in water and in ethanol (95). try <2.25>, and compare the spectrum with the Reference
It dissolves in sodium hydroxide TS with turbidity. Spectrum: both spectra exhibit similar intensities of absorp-
Identification (1) To 0.1 g of Albumin Tannate add 10
(2) To 0.2 g of Aldioxa add 10 mL of dilute hydrochloric
mL of ethanol (95), and heat in a water bath for 3 minutes
acid, dissolve by warming, and cool: the solution responds
with shaking. After cooling, filter, and to 5 mL of the fil-
to the Qualitative Tests <1.09> for aluminum salt.
trate add 1 drop of iron (III) chloride TS: a blue-purple to
bluish black color is produced. On standing, a bluish black Purity (1) Chloride <1.03>—To 0.10 g of Aldioxa add
precipitate is produced. 6 mL of dilute nitric acid, boil to dissolve with shaking for
(2) To 0.1 g of Albumin Tannate add 5 mL of nitric 5 minutes, cool, and add water to make 50 mL. Perform the
acid: an orange-yellow color develops. test using this solution as the test solution. Prepare the con-
trol solution with 0.40 mL of 0.01 mol/L hydrochloric acid
Purity (1) Acidity—Shake 1.0 g of Albumin Tannate with
VS (not more than 0.142z).
50 mL of water for 5 minutes, and filter. To 25 mL of the fil-
(2) Heavy metals <1.07>—To 1.0 g of Aldioxa add 3 mL
trate add 1.0 mL of 0.1 mol/L sodium hydroxide VS and
of hydrochloric acid and 3 mL of water, heat gently to boil
2 drops of phenolphthalein TS: a red color develops.
with shaking, and evaporate on a water bath to dryness. To
(2) Fats—To 2.0 g of Albumin Tannate add 20 mL of
the residue add 30 mL of water, shake under warming, cool,
petroleum benzine, shake vigorously for 15 minutes, and
filter, and to the filtrate add 2 mL of dilute acetic acid and
filter. Evaporate 10 mL of the filtrate on a water bath: the
water to make 50 mL. Perform the test using this solution as
mass of the residue is not more than 50 mg.
the test solution. Prepare the control solution as follows:
Loss on drying <2.41> Not more than 6.0z (1 g, 1059C, evaporate 3 mL of hydrochloric acid on a water bath to dry-
3 hours). ness, and add 2.0 mL of Standard Lead Solution, 2 mL of
dilute acetic acid and water to make 50 mL (not more than
20 ppm).
Digestion test To 1.00 g of Albumin Tannate add 0.25 g of
saccharated pepsin and 100 mL of water, shake well, and
2 hours).
allow to stand for 20 minutes at 40 ± 19 C in a water bath.
Add 1.0 mL of dilute hydrochloric acid, shake, and allow to Assay (1) Allantoin—Weigh accurately about 0.1 g of
stand for 3 hours at 40 ± 19C. Cool rapidly to ordinary tem- Aldioxa, previously dried, dissolve in 50 mL of dilute sulfu-
perature, and filter. Wash the residue with three 10-mL por- ric acid by heating, cool, and add water to make exactly 100
tions of water, dry in a desiccator (silica gel) for 18 hours, mL. Pipet 10 mL of this solution, and perform the test as
and dry at 1059C for 5 hours: the mass of the residue is 0.50 directed under Nitrogen Determination <1.08>.
to 0.58 g.
Each mL of 0.005 mol/L sulfuric acid VS
Containers and storage Containers—Tight containers. ＝ 0.3953 mg of C4H6N4O3
(2) Aluminum—Weigh accurately about 0.2 g of
Aldioxa, previously dried, dissolve carefully in 50 mL of
dilute hydrochloric acid by heating, cool, and add dilute
Aldioxa hydrochloric acid to make exactly 100 mL. Pipet 4 mL of
this solution, add water to make exactly 25 mL, and use this
アルジオキサ
solution as the sample solution. Separately, pipet a suitable
quantity of Standard Aluminum Stock Solution, dilute with
water so that each mL of the solution contains not less than
16.0 mg and not more than 64.0 mg of aluminum (Al: 26.98),
test with the sample solution and standard solution as di-
rected under Atomic Absorption Spectrophotometry <2.23>
according to the following conditions, and calculate the alu-
Dihydroxo[(4RS)-5-oxo-4-ureido-4,5-dihydro-1H-imidazol-
minum content of the sample solution from the calibration
2-yl]oxoaluminium
curve obtained from the absorbance of the standard solu-
[5579-81-7]
tion.
Gas: Combustible gas—Acetylene.
Aldioxa is a condensation product of allantoin and
Supporting gas—Nitrous oxide.
aluminum hydroxide.
Lamp: An aluminum hollow cathode lamp.
When dried, it contains not less than 65.3z and not
Wavelength: 309.2 nm.
more than 74.3z of allantoin (C4H6N4O3: 158.12),
JP XVII Official Monographs / Aldioxa Tablets 387
Containers and storage Containers—Well-closed contain- MS: Amount (mg) of aldioxa for assay taken
ers. MT: Amount (g) of Aldioxa Granules taken
C: Labeled amount (mg) of aldioxa (C4H7AlN4O5) in 1 g
Assay Weigh accurately an amount of powdered Aldioxa
Aldioxa Granules Granules, equivalent to about 0.1 g of aldioxa
(C4H7AlN4O5), add 80 mL of sodium fluoride-hydrochloric
アルジオキサ顆粒
acid TS, shake for 20 minutes, add sodium fluoride-
hydrochloric acid TS to make exactly 100 mL, and filter.
Aldioxa Granules contain not less than 95.0z and Pipet 2 mL of the filtrate, add diluted ammonia-ammonium
not more than 105.0z of the labeled amount of chloride buffer solution (pH 10.0) (1 in 10) to make exactly
aldioxa (C4H7AlN4O5: 218.10). 100 mL, and use this solution as the sample solution. Sepa-
rately, weigh accurately about 50 mg of aldioxa for assay,
Method of preparation Prepare as directed under Gran-
previously dried at 1059C for 2 hours, and dissolve in so-
ules, with Aldioxa.
dium fluoride-hydrochloric acid TS to make exactly 100 mL.
Identification (1) Determine the absorption spectrum of Pipet 4 mL of this solution, add diluted ammonia-
the sample solution obtained in the Assay as directed under ammonium chloride buffer solution (pH 10.0) (1 in 10) to
Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a make exactly 100 mL, and use this solution as the standard
maximum between 221 nm and 225 nm. solution. Determine the absorbances, AT and AS, at 223 nm
(2) To a quantity of powdered Aldioxa Granules, of the sample solution and standard solution as directed
equivalent to 0.2 g of Aldioxa, add 10 mL of dilute hydro- under Ultraviolet-visible Spectrophotometry <2.24>.
chloric acid, boil for 5 minutes, and filter: the cooled filtrate
Amount (mg) of aldioxa (C4H7AlN4O5)
responds to the Qualitative Tests <1.09> for aluminum salt.
＝ MS × AT/AS × 2
MS: Amount (mg) of aldioxa for assay taken
ing to the following method: Aldioxa Granules in single-dose
packages meet the requirement of the Content uniformity Containers and storage Containers—Tight containers.
test.
To the total content of 1 package of Aldioxa Granules add
80 mL of sodium fluoride-hydrochloric acid TS, shake for Aldioxa Tablets
20 minutes, add sodium fluoride-hydrochloric acid TS to
make exactly 100 mL, and filter. Pipet V mL of the filtrate, アルジオキサ錠
add diluted ammonia-ammonium chloride buffer solution
(pH 10.0) (1 in 10) to make exactly V? mL so that each mL
Aldioxa Tablets contain not less than 95.0z and
contains about 20 mg of aldioxa (C4H7AlN4O5), and use this
solution as the sample solution. Then, proceed as directed in
aldioxa (C4H7AlN4O5: 218.10).
the Assay.
with Aldioxa.
＝ MS × AT/AS × V?/V × 1/25
Dissolution <6.10> When the test is performed at 50 revolu- Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a
tions per minute according to the Paddle method, using 900 maximum between 221 nm and 225 nm.
Uniformity of dosage units <6.02> Perform the Mass varia-
in 15 minutes of Aldioxa Granules is not less than 85z.
tion test, or the Content uniformity test according to the fol-
Start the test with an accurately weighed amount of
lowing method: it meets the requirement.
Aldioxa Granules, equivalent to about 0.1 g of aldioxa
To 1 tablet of Aldioxa Tablets add 80 mL of sodium
(C4H7AlN4O5), withdraw not less than 20 mL of the medium
fluoride-hydrochloric acid TS, shake for 20 minutes, add so-
at the specified minute after starting the test, and filter
dium fluoride-hydrochloric acid TS to make exactly 100 mL,
through a membrane filter with a pore size not exceeding
and filter. Pipet V mL of the filtrate, add diluted ammonia-
0.45 mm. Discard the first 10 mL of the filtrate, pipet 10 mL
ammonium chloride buffer solution (pH 10.0) (1 in 10) to
of the subsequent filtrate, add diluted ammonia-ammonium
make exactly V? mL so that each mL contains about 20 mg of
chloride buffer solution (pH 10.0) (1 in 10) to make exactly
aldioxa (C4H7AlN4O5), and use this solution as the sample
50 mL, and use this solution as the sample solution. Sepa-
solution. Then, proceed as directed in the Assay.
rately, weigh accurately about 28 mg of aldioxa for assay,
previously dried at 1059C for 2 hours, and dissolve in so- Amount (mg) of aldioxa (C4H7AlN4O5)
dium fluoride-hydrochloric acid TS to make exactly 25 mL. ＝ MS × AT/AS × V?/V × 1/25
Pipet 1 mL of this solution, add diluted ammonia-
ammonium chloride buffer solution (pH 10.0) (1 in 10) to
make exactly 50 mL, and use this solution as the standard Dissolution <6.10> When the test is performed at 50 revolu-
solution. Determine the absorbances, AT and AS, at 223 nm tions per minute according to the Paddle method, using 900
of the sample solution and standard solution as directed mL of water as the dissolution medium, the dissolution rates
under Ultraviolet-visible Spectrophotometry <2.24>. in 15 minutes of 50-mg tablet and in 30 minutes of 100-mg
tablet are not less than 80z and not less than 70z, respec-
tively.
of aldioxa (C4H7AlN4O5)
Start the test with 1 tablet of Aldioxa Tablets, withdraw
＝ MS/MT × AT/AS × 1/C × 360
388 Alendronate Sodium Hydrate / Official Monographs JP XVII
after starting the test, and filter through a membrane filter white crystalline powder.
with a pore size not exceeding 0.45 mm. Discard the first 10 It is sparingly soluble in water, and practically insoluble in
mL of the filtrate, pipet V mL of the subsequent filtrate, add ethanol (99.5).
diluted ammonia-ammonium chloride buffer solution (pH It dissolves in 0.1 mol/L trisodium citrate TS.
10.0) (1 in 10) to make exactly V? mL so that each mL con- Melting point: about 2529C (with decomposition, after
tains about 22 mg of aldioxa (C4H7AlN4O5), and use this so- drying).
lution as the sample solution. Separately, weigh accurately
Identification (1) To 5 mL of a solution of Alendronate
about 28 mg of aldioxa for assay, previously dried at 1059C
Sodium Hydrate (1 in 50) add 1 mL of ninhydrin TS, and
for 2 hours, and dissolve in sodium fluoride-hydrochloric
heat: a blue-purple color develops.
acid TS to make exactly 25 mL. Pipet 1 mL of this solution,
(2) Determine the infrared absorption spectrum of Alen-
add diluted ammonia-ammonium chloride buffer solution
dronate Sodium Hydrate as directed in the potassium bro-
(pH 10.0) (1 in 10) to make exactly 50 mL, and use this solu-
mide disk method under Infrared Spectrophotometry <2.25>,
tion as the standard solution. Determine the absorbances, AT
and compare the spectrum with the Reference Spectrum or
and AS, at 223 nm of the sample solution and standard solu-
the spectrum of Alendronate Sodium RS: both spectra ex-
tion as directed under Ultraviolet-visible Spectrophotometry
hibit similar intensities of absorption at the same wave num-
<2.24>.
bers.
Dissolution rate (z) with respect to the labeled amount (3) To 0.1 g of Alendronate Sodium Hydrate add 10 mL
of aldioxa (C4H7AlN4O5) of a mixture of nitric acid and perchloric acid (1:1). Heat to
＝ MS × AT/AS × V?/V × 1/C × 72 concentrate to about 1 mL, add about 10 mL of water while
hot, and neutralize with a solution of sodium hydroxide (2 in
5): the solution responds to the Qualitative Tests <1.09> for
C: Labeled amount (mg) of aldioxa (C4H7AlN4O5) in 1
phosphate.
tablet
(4) A solution of Alendronate Sodium Hydrate (1 in 100)
Assay Weigh accurately, and powder not less than 20 responds to the Qualitative Tests <1.09> for sodium salt.
Aldioxa Tablets. Weigh accurately a portion of the powder,
pH <2.54> The pH of a solution of 1.0 g of Alendronate
equivalent to about 0.1 g of aldioxa (C4H7AlN4O5), add 80
Sodium Hydrate in 100 mL of freshly boiled and cooled
mL of sodium fluoride-hydrochloric acid TS, shake for 20
water is between 4.0 and 5.0.
minutes, add sodium fluoride-hydrochloric acid TS to make
exactly 100 mL, and filter. Pipet 2 mL of the filtrate, add Purity (1) Heavy metals <1.07>—Put 1.0 g of Alen-
diluted ammonia-ammonium chloride buffer solution (pH dronate Sodium Hydrate in a Kjeldahl flask, add 9 mL of a
10.0) (1 in 10) to make exactly 100 mL, and use this solution mixture of nitric acid and sulfuric acid (5:4), and heat until
as the sample solution. Separately, weigh accurately about the solution becomes brown. After cooling, add 9 mL of a
50 mg of aldioxa for assay, previously dried at 1059C for 2 mixture of nitric acid and sulfuric acid (5:4), and heat again
hours, and dissolve in sodium fluoride-hydrochloric acid TS until the color changes from colorless to brown. After cool-
to make exactly 100 mL. Pipet 4 mL of this solution, add ing, add 2 mL of nitric acid, strongly heat until brown fumes
diluted ammonia-ammonium chloride buffer solution (pH are no longer evolved, and heat until large amounts of white
10.0) (1 in 10) to make exactly 100 mL, and use this solution fumes are evolved. After cooling, add carefully 5 mL of
as the standard solution. Determine the absorbances, AT and water and 1 mL of hydrogen peroxide (30), heat until white
AS, at 223 nm of the sample solution and standard solution fumes are no longer evolved, and continue heating for more
as directed under Ultraviolet-visible Spectrophotometry 5 minutes. After cooling, if any yellow color remains, add 2
<2.24>. mL of nitric acid, and repeat the same procedure. After
cooling, transfer the solution in the Kjeldahl flask to a
beaker, wash out the inside of the flask with 5 mL of water,
＝ M S × AT / AS × 2
and add the washing to the beaker. Adjust to pH 3 – 5 with
MS: Amount (mg) of aldioxa for assay taken ammonia solution (28), transfer to a Nessler tube, add water
to make 50 mL, and perform the test with this solution as the
test solution. Prepare the control solution in the same proce-
dure using the same amount of the reagents used for the
preparation of the sample solution, add 1.0 mL of Standard
Alendronate Sodium Hydrate Lead Solution and add water to make 50 mL (not more than
10 ppm).
アレンドロン酸ナトリウム水和物
(2) Related substances—Dissolve 15 mg of Alendronate
Sodium Hydrate in 25 mL of 0.1 mol/L trisodium citrate
TS, and use this solution as the sample stock solution. Pipet
5 mL of the sample stock solution, and add 0.1 mol/L triso-
dium citrate TS to make exactly 50 mL. Pipet 1 mL of this
C4H12NNaO7P2.3H2O: 325.12
solution, add 0.1 mol/L trisodium citrate TS to make exactly
Monosodium trihydrogen 4-amino-1-hydroxybutane-
100 mL, and use this solution as the standard stock solution.
1,1-diyldiphosphonate trihydrate
To exactly 5 mL each of the sample stock solution and
[121268-17-5]
standard stock solution, add exactly 5 mL each of a solution
of sodium tetraborate decahydrate (19 in 1000), acetonitrile
Alendronate Sodium Hydrate contains not less than
and a solution of 9-fluorenylmethyl chloroformate in aceto-
99.0z and not more than 101.0z of alendronate so-
nitrile (1 in 250), shake for 45 seconds, and allow to stand
dium (C4H12NNaO7P2: 271.08), calculated on the dried
for 30 minutes at room temperature. Then, add 20 mL of
basis.
dichloromethane to them, shake for 60 seconds, centrifuge,
Description Alendronate Sodium Hydrate occurs as a and use the supernatant liquids so obtained as the sample
JP XVII Official Monographs / Alendronate Sodium Injection 389
solution and the standard solution, respectively. Perform decahydrate (19 in 1000) and a solution of 9-fluorenylmethyl
the test with exactly 20 mL each of the sample solution and chloroformate in acetonitrile (1 in 2000), shake for 30
standard solution as directed under Liquid Chromatography seconds, and allow to stand for 25 minutes. Then, add 25
<2.01> according to the following conditions. Determine each mL of dichloromethane, shake for 60 seconds, centrifuge,
peak area of these solutions by the automatic integration and use the supernatant liquids so obtained as the sample
method: each peak area other than alendronic acid obtained solution and the standard solution, respectively. Perform
from the sample solution is not larger than the peak area of the test with exactly 10 mL each of the sample solution and
alendronic acid obtained from the standard solution. standard solution as directed under Liquid Chromatography
Operating conditions— <2.01> according to the following conditions, and determine
Detector: An ultraviolet absorption photometer (wave- the peak areas, AT and AS, of alendronic acid in each solu-
length: 266 nm). tion.
Amount (mg) of alendronate sodium (C4H12NNaO7P2)
ter and 25 cm in length, packed with styrene-divinylbenzene
＝ MS × AT/AS
copolymer for liquid chromatography (10 mm in particle di-
ameter). MS: Amount (mg) of Alendronate Sodium RS taken, cal-
Column temperature: A constant temperature of about culated on the dried basis
459 C.
Mobile phase A: Dissolve 2.94 g of trisodium citrate dihy-
drate and 1.42 g of anhydrous disodium hydrogen phosphate
length: 266 nm).
in 900 mL of water, adjust to pH 8.0 with phosphoric acid,
and add water to make 1000 mL. To 850 mL of this solution
add 150 mL of acetonitrile for liquid chromatography.
Mobile phase B: Dissolve 2.94 g of trisodium citrate dihy-
ameter).
359C.
Mobile phase: Dissolve 14.7 g of trisodium citrate dihy-
add 700 mL of acetonitrile for liquid chromatography.
Flowing of mobile phase: Control the gradient by mixing
the mobile phases A and B as directed in the following table.
add 250 mL of acetonitrile for liquid chromatography and
Time after injection Mobile phase A Mobile phase B 50 mL of methanol.
of sample (min) (volz) (volz) Flow rate: Adjust so that the retention time of alendronic
acid is about 3 minutes.
0 – 15 100 → 50 0 → 50
15 – 25 50 → 0 50 → 100
Flow rate: About 1.8 mL per minute. ditions, the number of theoretical plates and the symmetry
Time span of measurement: About 5 times as long as the factor of the peak of alendronic acid are not less than 1500
retention time of alendronic acid, beginning after the solvent and not more than 1.5, respectively.
peak. System repeatability: When the test is repeated 6 times
System suitability— with 10 mL of the standard solution under the above operat-
System performance: Dissolve 15 mg of Alendronate So- ing conditions, the relative standard deviation of the peak
dium Hydrate and 2 mg of 4-aminobutylic acid in 100 mL of area of alendronic acid is not more than 1.0z.
0.1 mol/L trisodium citrate TS. To 5 mL of this solution add
5 mL each of a solution of sodium tetraborate decahydrate Containers and storage Containers—Tight containers.
(19 in 1000), acetonitrile and a solution of 9-fluorenylmethyl
chloroformate in acetonitrile (1 in 250), then, proceed in the
same manner as the sample solution. When the procedure is Alendronate Sodium Injection
run with 20 mL of this solution under the above operating
conditions, alendronic acid and 4-aminobutylic acid are アレンドロン酸ナトリウム注射液
eluted in this order with the resolution between these peaks
being not less than 6. Alendronate Sodium Injection is an aqueous injec-
System repeatability: When the test is repeated 6 times tion.
with 20 mL of the standard solution under the above operat- It contains not less than 95.0z and not more than
ing conditions, the relative standard deviation of the peak 105.0z of the labeled amount of alendronic acid
area of alendronic acid is not more than 2.0z. (C4H13NO7P2: 249.10).
Loss on drying <2.41> 16.1 – 17.1z (1 g, 1409C, 3 hours). Method of preparation Prepare as directed under Injec-
Assay Weigh accurately about 10 mg each of Alendronate tions, with Alendronate Sodium Hydrate.
Sodium Hydrate and Alendronate Sodium RS (separately Description Alendronate Sodium Injection is a clear, color-
determine the loss on drying <2.41> in the same conditions as less liquid.
Alendronate Sodium Hydrate), dissolve in 0.1 mol/L triso-
dium citrate TS to make exactly 100 mL, and use these solu- Identification Use Alendronate Sodium Injection as the
tions as the sample stock solution and the standard stock so- sample solution. Separately, dissolve 33 mg of alendronate
lution, respectively. To exactly 5 mL each of these solutions sodium hydrate in 10 mL of water, and use this solution as
add exactly 5 mL each of a solution of sodium tetraborate the standard solution. Perform the test with these solutions
as directed under Thin-layer Chromatography <2.03>. Spot
390 Alendronate Sodium Tablets / Official Monographs JP XVII
5 mL each of the sample solution and standard solution on a 200 mL of acetonitrile for liquid chromatography and 50 mL
plate of cellulose for thin-layer chromatography. Develop of methanol.
the plate with a mixture of water, pyridine, acetic acid (100) Flow rate: Adjust so that the retention time of alendronic
and ethyl acetate (1:1:1:1) to a distance of about 10 cm, and acid is about 7 minutes.
air-dry the plate. Spray evenly a solution of ninhydrin in ace- System suitability—
tone (1 in 50) on the plate, and heat at 1009C for 10 minutes: System performance: When the procedure is run with 50
the principal spots from the sample solution and standard mL of the standard solution under the above operating con-
solution show a blue-purple color and the same R f value. ditions, the number of theoretical plates and the symmetry
factor of the peak of alendronic acid are not less than 1500
pH Being specified separately when the drug is granted ap-
and not more than 1.5, respectively.
proval based on the Law.
Bacterial endotoxins <4.01> Less than 119 EU/mg. with 50 mL of the standard solution under the above opera-
tions conditions, the relative standard deviation of the peak
of alendronic acid is not more than 1.0z.
ment. Alendronate Sodium Tablets
Sterility <4.06> Perform the test according to Membrane-
アレンドロン酸ナトリウム錠
filter method: it meets the requirement.
Assay To an exactly measured volume of Alendronate So-
Alendronate Sodium Tablets contain not less than
dium Injection, equivalent to about 5 mg of alendronic acid
95.0z and not more than 105.0z of the labeled
(C4H13NO7P2), add a solution of disodium dihydrogen ethyl-
amount of alendronic acid (C4H13NO7P2: 249.10).
enediamine tetraacetate dihydrate (1 in 500) to make exactly
100 mL, and use this solution as the sample stock solution. Method of preparation Prepare as directed under Tablets,
Separately, weigh accurately about 33 mg of Alendronate with Alendronate Sodium Hydrate.
Sodium RS (separately determine the loss on drying <2.41>
Identification To a quantity of powdered Alendronate
under the same conditions as Alendronate Sodium Hydrate),
Sodium Tablets, equivalent to 25 mg of alendronic acid
and dissolve in a solution of disodium dihydrogen ethylene-
(C4H13NO7P2), add 25 mL of water, shake, centrifuge, and
diamine tetraacetate dihydrate (1 in 500) to make exactly 100
use the supernatant liquid as the sample solution. Separately,
mL. Pipet 10 mL of this solution, add a solution of diso-
weigh 33 mg of alendronate sodium hydrate, and dissolve in
dium dihydrogen ethylenediamine tetraacetate dihydrate (1
25 mL of water, and use this solution as the standard solu-
in 500) to make exactly 50 mL, and use this solution as the
tion. Perform the test with these solutions as directed under
standard stock solution. Pipet 5 mL each of the sample stock
Thin-layer Chromatography <2.03>. Spot 5 mL each of the
solution and standard stock solution, add exactly 5 mL of a
sample solution and standard solution on a plate of cellulose
solution of sodium tetraborate decahydrate (19 in 500) and
for thin-layer chromatography. Develop the plate with a
exactly 4 mL of a solution of 9-fluorenylmethyl chlorofor-
mixture of water, pyridine, acetic acid (100) and ethyl ace-
mate in acetonitrile (1 in 1000), shake for 30 seconds, and
tate (1:1:1:1) to a distance of about 10 cm, and air-dry the
allow to stand at room temperature for 25 minutes. Then,
plate. Spray evenly a solution of ninhydrin in acetone (1 in
add 25 mL of dichloromethane to them, shake for 45
50) on the plate, and heat at 1009C for 10 minutes: the prin-
seconds, centrifuge, and use the supernatant liquid so ob-
cipal spots from the sample solution and standard solution
tained as the sample solution and the standard solution,
show a blue-purple color and the same R f value.
respectively. Perform the test with 50 mL each of the sample
solution and standard solution as directed under Liquid Uniformity of dosage units <6.02> Perform the Mass varia-
Chromatography <2.01> according to the following condition test, or the Content uniformity test according to the fol-
tions, and determine the peak areas, AT and AS, of alen- lowing method: it meets the requirement.
dronic acid in each solution. To 1 tablet of Alendronate Sodium Tablets add 0.1 mol/L
trisodium citrate TS to make exactly 100 mL, and stir until
Amount (mg) of alendronic acid (C4H13NO7P2)
the tablet is completely disintegrated. Centrifuge this solu-
＝ MS × AT/AS × 1/5 × 0.919
tion, pipet V mL of the supernatant liquid, and add
MS: Amount (mg) of Alendronate Sodium RS taken, cal- 0.1 mol/L trisodium citrate TS to make exactly V? mL so
culated on the dried basis that each mL contains about 25 mg of alendronic acid
(C4H13NO7P2), and use this solution as the sample stock
length: 265 nm). Amount (mg) of alendronic acid (C4H13NO7P2)
Column: A stainless steel column 4.1 mm in inside diame- ＝ MS × AT/AS × V?/V × 2/25 × 0.919
MS: Amount (mg) of Alendronate Sodium RS taken, cal-
culated on the dried basis
ameter).
Column temperature: A constant temperature of about Dissolution <6.10> When the test is performed at 50 revolu-
409 C. tions per minute according to the Paddle method, using 900
Mobile phase: Dissolve 14.7 g of trisodium citrate dihy- mL of water as the dissolution medium, the dissolution rate
drate and 8.7 g of dipotassium hydrogen phosphate in 900 in 15 minutes of Alendronate Sodium Tablets is not less than
mL of water, adjust to pH 8.0 with phosphoric acid, and 85z.
add water to make 1000 mL. To 750 mL of this solution add Start the test with 1 tablet of Alendronate Sodium Tablets,
JP XVII Official Monographs / Alimemazine Tartrate 391
withdraw not less than 10 mL of the medium at the specified Column: A stainless steel column 4.1 mm in inside diame-
minute after starting the test, and centrifuge. Pipet V mL ter and 25 cm in length, packed with styrene-divinylbenzene
of the supernatant liquid, add water to make exactly V? mL copolymer for liquid chromatography (10 mm in particle di-
so that each mL contains about 6 mg of alendronic acid ameter).
(C4H13NO7P2), and use this solution as the sample stock so- Column temperature: A constant temperature of about
lution. Separately, weigh accurately about 29 mg of Alen- 359C.
dronate Sodium RS (separately determine the loss on drying Mobile phase: Dissolve 14.7 g of trisodium citrate dihy-
<2.41> under the same conditions as Alendronate Sodium drate and 7.1 g of anhydrous disodium hydrogen phosphate
Hydrate), and dissolve in water to make exactly 250 mL. in 900 mL of water, adjust to pH 8.0 with phosphoric acid,
Pipet 3 mL of this solution, add water to make exactly 50 and add water to make 1000 mL. To 750 mL of this solution
mL, and use this solution as the standard stock solution. add 200 mL of acetonitrile for liquid chromatography and
Pipet 5 mL each of the sample stock solution and standard 50 mL of methanol.
stock solution, add exactly 1 mL of trisodium citrate dihy- Flow rate: Adjust so that the retention time of alendronic
drate solution (22 in 125), exactly 5 mL of a solution ob- acid is about 7 minutes.
tained by dissolving 6.2 g of boric acid in 950 mL of water, System suitability—
adjusting to pH 9.0 with sodium hydrate TS, and adding System performance: When the procedure is run with 50
water to make 1000 mL, and add exactly 4 mL of a solution mL of the standard solution under the above operating con-
of 9-fluorenylmethyl chloroformate in acetonitrile (1 in ditions, the number of theoretical plates and the symmetry
2000), shake for 30 seconds, and allow to stand at room factor of the peak of alendronic acid are not less than 1500
temperature for 25 minutes. Add 25 mL of dichloromethane, and not more than 1.5, respectively.
shake for 45 seconds, then centrifuge, and use the superna- System repeatability: When the test is repeated 6 times
tant liquid as the sample solution and the standard solution, with 50 mL of the standard solution under the above opera-
respectively. Then, proceed as directed in the Assay. tions conditions, the relative standard deviation of the peak
of alendronic acid is not more than 1.0z.
of alendronic acid (C4H13NO7P2) Containers and storage Containers—Well-closed contain-
＝ MS × AT/AS × V?/V × 1/C × 108/5 × 0.919 ers.
C: Labeled amount (mg) of alendronic acid (C4H13NO7P2) Alimemazine Tartrate
in 1 tablet
アリメマジン酒石酸塩
Assay Weigh accurately and powder not less than 20
Alendronate Sodium Tablets. Weigh accurately a portion of
the powder, equivalent to about 50 mg of alendronic acid
(C4H13NO7P2), add 0.1 mol/L trisodium citrate TS to make
exactly 1000 mL, stir for 30 minutes, and centrifuge. Pipet
5 mL of the supernatant liquid, add 0.1 mol/L trisodium
citrate TS to make exactly 10 mL, and use this solution as
the sample stock solution. Separately, weigh accurately
(C18H22N2S)2.C4H6O6: 746.98
about 39 mg of Alendronae Sodium RS (separately deter-
N, N,2-Trimethyl-3-(10H-phenothiazin-10-
mine the loss on drying <2.41> under the same conditions as
yl)propylamine hemitartrate
Alendronae Sodium Hydrate), dissolve in 0.1 mol/L triso-
[41375-66-0]
dium citrate TS to make exactly 50 mL. Pipet 2 mL of this
solution, add 0.1 mol/L trisodium citrate TS to make exactly
Alimemazine Tartrate, when dried, contains not less
50 mL, and use this solution as the standard stock solution.
than 98.0z of alimemazine tartrate [(C18H22N2S)2.
Pipet 5 mL each of the sample stock solution and standard
C4H6O6].
stock solution, add exactly 5 mL of a solution of sodium
tetraborate decahydrate (19 in 500) and exactly 4 mL of a so- Description Alimemazine Tartrate occurs as a white pow-
lution of 9-fluprenylmethyl chloroformate in acetonitrile (1 der. It is odorless, and has a bitter taste.
in 1000), shake for 30 seconds, and allow to stand at room It is freely soluble in water and in acetic acid (100), spar-
temperature for 25 minutes. Then, add 25 mL of dichloro- ingly soluble in ethanol (95), and practically insoluble in
methane to them, shake for 45 seconds, centrifuge, and use diethyl ether.
the supernatant liquid so obtained as the sample solution The pH of a solution of 1.0 g of Alimemazine Tartrate in
and the standard solution, respectively. Perform the test 50 mL of water is between 5.0 and 6.5.
with exactly 50 mL each of the sample solution and standard It is gradually colored by light.
Identification (1) To 2 mL of a solution of Alimemazine
according to the following conditions, and determine the
Tartrate (1 in 100) add 1 drop of iron (III) chloride TS: a
peak areas, AT and AS, of alendronic acid in each solution.
red-brown color is produced, and immediately a yellow pre-
Amount (mg) of alendronic acid (C4H13NO7P2) cipitate is formed.
＝ MS × AT/AS × 8/5 × 0.919 (2) Dissolve 1 g of Alimemazine Tartrate in 5 mL of
water, add 3 mL of sodium hydroxide TS, extract with two
10-mL portions of diethyl ether [use the aqueous layer ob-
tained in the Identification (4)]. Shake the combined diethyl
Operating conditions— ether extracts with 3 g of anhydrous sodium sulfate, filter,
Detector: An ultraviolet absorption photometer (wave- and evaporate the diethyl ether with the aid of a current of
length: 266 nm). air. Dry the residue in a desiccator (in vacuum, phosphorus
392 Allopurinol / Official Monographs JP XVII
(V) oxide) for 16 hours: it melts <2.60> between 669C and similar intensities of absorption at the same wavelengths.
709 C. (2) Determine the infrared absorption spectrum of
(3) Determine the absorption spectrum of a solution of Allopurinol, previously dried, as directed in the potassium
Alimemazine Tartrate (1 in 100,000) as directed under Ultra- bromide disk method under Infrared Spectrophotometry
violet-visible Spectrophotometry <2.24>, and compare the <2.25>, and compare the spectrum with the Reference Spec-
spectrum with the Reference Spectrum: both spectra exhibit trum: both spectra exhibit similar intensities of absorption at
similar intensities of absorption at the same wavelengths. the same wave numbers.
(4) The aqueous layer, obtained in the identification (2),
when neutralized with dilute acetic acid, responds to the
Allopurinol according to Method 2, and perform the test.
Qualitative Tests <1.09> (1) and (2) for tartrate.
Melting point <2.60> 159 – 1639C Solution (not more than 20 ppm).
of Allopurinol according to Method 3, and perform the test
of Alimemazine Tartrate in 20 mL of water: the solution is
(not more than 2 ppm).
clear and colorless.
(3) Related substances—Dissolve 50 mg of Allopurinol
in 10 mL of ammonia TS, and use this solution as the sample
Alimemazine Tartrate according to Method 2, and perform
solution. Pipet 1 mL of the sample solution, add ammonia
the test. Prepare the control solution with 2.0 mL of Stand-
TS to make exactly 500 mL, and use this solution as the
directed under Thin-layer Chromatography <2.03>. Spot 5
of Alimemazine Tartrate according to Method 3, and per-
mL each of the sample solution and standard solution on a
form the test. Use a solution of magnesium nitrate hexahy-
plate of cellulose with fluorescent indicator for thin-layer
drate in ethanol (95) (1 in 5) (not more than 2 ppm).
chromatography. Develop the plate with ammonia TS-satu-
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, rated 1-butanol to a distance of about 10 cm, and air-dry the
3 hours). plate. Examine under ultraviolet light (main wavelength: 254
nm): the spots other than the principal spot from the sample
solution are not more intense than the spot from the stand-
Assay Weigh accurately about 0.8 g of Alimemazine Tar- ard solution.
trate, previously dried, dissolve in 50 mL of acetic acid (100),
and titrate <2.50> with 0.1 mol/L perchloric acid VS until the
4 hours).
color of the solution changes from red through brown to
green-brown (indicator: 2 mL of p-naphtholbenzein TS). Residue on ignition <2.44> Not more than 0.1z (1 g).
Assay Weigh accurately about 0.16 g of Allopurinol, previ-
correction.
ously dried, dissolve in 70 mL of N, N-dimethylformamide
Each mL of 0.1 mol/L perchloric acid VS by warming. Cool, and titrate <2.50> with 0.1 mol/L tetra-
＝ 37.35 mg of (C18H22N2S)2.C4H6O6 methylammonium hydroxide VS (potentiometric titration).
To 70 mL of N, N-dimethylformamide add 12 mL of water,
perform a blank determination, and make any necessary cor-
rection.
Each mL of 0.1 mol/L tetramethylammonium
Allopurinol hydroxide VS
＝ 13.61 mg of C5H4N4O
アロプリノール
Allopurinol Tablets
アロプリノール錠
C5H4N4O: 136.11
1H-Pyrazolo[3,4-d ]pyrimidin-4-ol
Allopurinol Tablets contain not less than 93.0z
[315-30-0]
allopurinol (C5H4N4O: 136.11).
Allopurinol, when dried, contains not less than
98.0z and not more than 101.0z of allopurinol Method of preparation Prepare as directed under Tablets,
(C5H4N4O). with Allopurinol.
Description Allopurinol occurs as white to pale yellowish Identification (1) Determine the absorption spectrum of
white, crystals or crystalline powder. the sample solution obtained in the Assay as directed under
It is slightly soluble in N, N-dimethylformamide, and very Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a
slightly soluble in water and in ethanol (99.5). maximum between 248 nm and 252 nm.
It dissolves in ammonia TS. (2) To a quantity of powdered Allopurinol Tablets,
equivalent to 0.1 g of Allopurinol, add 5 mL of a solution of
diethylamine (1 in 10), shake well, add 5 mL of methanol,
solution of Allopurinol (1 in 200,000) as directed under Ul-
centrifuge, and use the supernatant liquid as the sample solu-
traviolet-visible Spectrophotometry <2.24>, and compare the
tion. Separately, dissolve 0.1 g of allopurinol in 5 mL of a
JP XVII Official Monographs / Alminoprofen 393
solution of diethylamine (1 in 10), add 5 mL of methanol, Allopurinol Tablets, and powder. Weigh accurately a por-
and use this solution as the standard solution. Perform the tion of the powder, equivalent to about 0.1 g of allopurinol
test with these solutions as directed under Thin-layer Chro- (C5H4N4O), add 20 mL of 0.05 mol/L sodium hydroxide TS,
matography <2.03>. Spot 2.5 mL each of the sample solution shake well, and treat with ultrasonic waves for 10 minutes.
and standard solution on a plate of silica gel with fluorescent After cooling, add 0.1 mol/L hydrochloric acid TS to make
indicator for thin-layer chromatography. Develop the plate exactly 200 mL, and filter through a membrane filter with a
with a mixture of 2-butanone, ammonia solution (28) and 2- pore size not exceeding 0.8 mm. Discard the first 10 mL of
methoxyethanol (3:1:1) to a distance of about 10 cm, and the filtrate, pipet 2 mL of the subsequent filtrate, add 0.1
air-dry the plate. Examine under ultraviolet light (main mol/L hydrochloric acid TS to make exactly 100 mL, and
wavelength: 254 nm): the principal spots obtained from the use this solution as the sample solution. Separately, weigh
sample solution and standard solution show the same R f accurately about 0.1 g of allopurinol for assay, previously
value. dried at 1059C for 4 hours, dissolve in 20 mL of 0.05 mol/L
sodium hydroxide TS, and add 0.1 mol/L hydrochloric acid
TS to make exactly 200 mL. Pipet 2 mL of this solution, add
0.1 mol/L hydrochloric acid TS to make exactly 100 mL,
and use this solution as the standard solution. Determine the
To 1 tablet of Allopurinol Tablets add V/10 mL of 0.05
absorbances, AT and AS, of the sample solution and stand-
mol/L sodium hydroxide TS, shake well, and treat with
ard solution at 250 nm as directed under Ultraviolet-visible
ultrasonic waves for 10 minutes. After cooling, add 0.1
Spectrophotometry <2.24>.
mol/L hydrochloric acid TS to make exactly V mL so that
each mL contains about 0.5 mg of allopurinol (C5H4N4O), Amount (mg) of allopurinol (C5H4N4O) ＝ MS × AT/AS
and filter through a membrane filter with a pore size not
MS: Amount (mg) of allopurinol for assay taken
pipet 2 mL of the subsequent filtrate, add 0.1 mol/L of hy- Containers and storage Containers—Well-closed contain-
drochloric acid TS to make exactly 100 mL, and use this so- ers.
about 50 mg of allopurinol for assay, previously dried at
1059C for 4 hours, dissolve in 10 mL of 0.05 mol/L sodium Alminoprofen
hydroxide TS, and add 0.1 mol/L hydrochloric acid TS to
make exactly 100 mL. Pipet 2 mL of this solution, add 0.1 アルミノプロフェン
mol/L hydrochloric acid TS to make exactly 100 mL, and
use this solution as the standard solution. Determine the ab-
sorbances, AT and AS, of the sample solution and standard
solution at 250 nm as directed under Ultraviolet-visible Spec-
trophotometry <2.24>.
Amount (mg) of allopurinol (C5H4N4O)
C13H17NO2: 219.28
＝ MS × AT/AS × V/100
(2RS )-2-{[4-(2-Methylprop-2-en-1-
MS: Amount (mg) of allopurinol for assay taken yl)amino]phenyl}propanoic acid
[39718-89-3]
Alminoprofen, when dried, contains not less than
99.0z and not more than 101.0z of alminoprofen
in 30 minutes of Allopurinol Tablets is not less than 80z.
(C13H17NO2).
Start the test with 1 tablet of Allopurinol Tablets, with-
draw not less than 20 mL of the medium at the specified Description Alminoprofen occurs as white to pale yellow,
minute after starting the test, and filter through a membrane crystals or crystalline powder.
filter with a pore size not exceeding 0.8 mm. Discard the first It is freely soluble in ethanol (99.5) and in acetic acid
10 mL of the filtrate, pipet V mL of the subsequent filtrate, (100), and very slightly soluble in water.
add water to make exactly V? mL so that each mL contains It gradually turns brown on exposure to light.
about 11 mg of allopurinol (C5H4N4O), and use this solution A solution of Alminoprofen in ethanol (99.5) (1 in 10)
as the sample solution. Separately, weigh accurately about shows no optical rotation.
11 mg of allopurinol for assay, previously dried at 1059C for
4 hours, and dissolve in water to make exactly 100 mL. Pipet
solution of Alminoprofen in ethanol (99.5) (3 in 500,000) as
5 mL of this solution, add water to make exactly 50 mL, and
and compare the spectrum with the Reference Spectrum:
both spectra exhibit similar intensities of absorption at the
same wavelengths.
Dissolution rate (z) with respect to the labeled amount Alminoprofen as directed in the potassium bromide disk
of allopurinol (C5H4N4O) method under Infrared Spectrophotometry <2.25>, and com-
＝ MS × AT/AS × V?/V × 1/C × 90 pare the spectrum with the Reference Spectrum: both spectra
exhibit similar intensities of absorption at the same wave
MS: Amount (mg) of allopurinol for assay taken
numbers.
C: Labeled amount (mg) of allopurinol (C5H4N4O) in 1
tablet Melting point <2.60> 106 – 1089C
Assay Weigh accurately the mass of not less than 20 Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
394 Alminoprofen Tablets / Official Monographs JP XVII
Alminoprofen according to Method 2, and perform the test. Containers and storage Containers—Well-closed contain-
Prepare the control solution with 2.0 mL of Standard Lead ers.
Solution (not more than 10 ppm). Storage—Light-resistant.
of Alminoprofen according to Method 3, and perform the
test (not more than 2 ppm). Alminoprofen Tablets
(3) Related substances—Conduct this procedure using
light-resistant vessels. Dissolve 50 mg of Alminoprofen in アルミノプロフェン錠
100 mL of the mobile phase, and use this solution as the
sample solution. Pipet 2 mL of the sample solution, add the
Alminoprofen Tablets contain not less than 93.0z
alminoprofen (C13H17NO2: 219.28).
under Liquid Chromatography <2.01> according to the fol- Method of preparation Prepare as directed under Tablets,
lowing conditions. Determine each peak area of these solu- with Alminoprofen.
tions by the automatic integration method: the area of the
Identification Take an amount of powdered Alminoprofen
peak other than alminoprofen obtained from the sample so-
Tablets, equivalent to 30 mg of Alminoprofen, add ethanol
lution is not larger than 1/5 times the peak area of almino-
(99.5) to make 100 mL, shake thoroughly, and centrifuge.
profen obtained from the standard solution. Furthermore,
To 2 mL of the supernatant liquid add ethanol (99.5) to
the total area of the peaks other than alminoprofen from the
make 100 mL, and determine the absorption spectrum of
sample solution is not larger than the peak area of almino-
this solution as directed under Ultraviolet-visible Spectro-
profen from the standard solution.
photometry <2.24>: it exhibits maxima between 253 nm and
257 nm, and between 298 nm and 302 nm.
length: 254 nm). Purity Related substances—Conduct this procedure using
Column: A stainless steel column 6.0 mm in inside diame- light-resistant vessels. Powder 10 tablets of Alminoprofen
ter and 15 cm in length, packed with octadecylsilanized silica Tablets, weigh a portion of the powder equivalent to 50 mg
gel for liquid chromatography (5 mm in particle diameter). of Alminoprofen, add 50 mL of the mobile phase, shake for
Column temperature: A constant temperature of about 15 minutes, add the mobile phase to make exactly 100 mL,
259 C. centrifuge, and use the supernatant liquid as the sample solu-
Mobile phase: A mixture of methanol and diluted acetic tion. Pipet 2 mL of the sample solution, add the mobile
acid (100) (1 in 1000) (4:1). phase to make exactly 200 mL, and use this solution as the
Flow rate: Adjust so that the retention time of almino- standard solution. Perform the test with exactly 5 mL each of
profen is about 5 minutes. the sample solution and standard solution as directed under
Time span of measurement: About 5 times as long as the Liquid Chromatography <2.01> according to the following
retention time of alminoprofen, beginning after the solvent conditions. Determine each peak area of each solution by the
peak. automatic integration method: the area of the peak other
System suitability— than alminoprofen obtained from the sample solution is not
Test for required detectability: Pipet 1 mL of the standard larger than 1/2 times the peak area of alminoprofen ob-
solution, and add the mobile phase to make exactly 10 mL. tained from the standard solution. Furthermore, the total
Confirm that the peak area of alminoprofen obtained from area of the peaks other than alminoprofen from the sample
5 mL of this solution is equivalent to 7 to 13z of that ob- solution is not larger than 2 times the peak area of almino-
tained from 5 mL of the standard solution. profen from the standard solution.
System performance: Dissolve 10 mg each of Almino- Operating conditions—
profen and butyl parahydroxybenzoate in 100 mL of metha- Proceed as directed in the operating conditions in the
nol. Pipet 10 mL of this solution, and add methanol to make Purity (3) under Alminoprofen.
exactly 50 mL. When the procedure is run with 5 mL of this System suitability—
solution under the above operating conditions, alminoprofen Proceed as directed in the system suitability in the Purity
and butyl parahydroxybenzoate are eluted in this order with (3) in Assay under Alminoprofen.
the resolution between these peaks being not less than 2.0.
To 1 tablet of Alminoprofen Tablets add 5 mL of water,
of alminoprofen is not more than 2.0z.
shake until the tablet is disintegrated, add 50 mL of ethanol
Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- (99.5), shake for 20 minutes, then add ethanol (99.5) to
um, phosphorus (V) oxide, 1 hour). make exactly 100 mL, and centrifuge. Pipet 3 mL of the su-
pernatant liquid, add ethanol (99.5) to make exactly 50 mL.
Pipet V mL of this solution, add ethanol (99.5) to make
Assay Weigh accurately about 0.3 g of Alminoprofen, exactly V? mL so that each mL contains about 6 mg of
previously dried, dissolve in 50 mL of acetic acid (100), and alminoprofen (C13H17NO2), and use this solution as the sam-
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- ple solution. Then, proceed as directed in the Assay.
metric titration). Perform a blank determination in the same
Amount (mg) of alminoprofen (C13H17NO2)
＝ MS × AT/AS × V?/V × 1/3
MS: Amount (mg) of alminoprofen for assay taken
＝ 21.93 mg of C13H17NO2
JP XVII Official Monographs / Alprazolam 395

mL of 2nd fluid for dissolution test as the dissolution me- Alprazolam
dium, the dissolution rate in 45 minutes of Alminoprofen
Tablets is not less than 80z. アルプラゾラム
Start the test with 1 tablet of Alminoprofen Tablets, with-
draw not less than 20 mL of the medium at specified minute
0.05 mol/L sodium hydroxide TS to make exactly V? mL so
that each mL contains about 8.9 mg of alminoprofen
(C13H17NO2), and use this solution as the sample solution.
Separately, weigh accurately about 30 mg of alminoprofen
C17H13ClN4: 308.76
for assay, previously dried in vacuum for 1 hour using phos-
8-Chloro-1-methyl-6-phenyl-4H-
phorus (V) oxide as the dessicant, and dissolve in 0.05 mol/L
[1,2,4]triazolo[4,3-a][1,4]benzodiazepine
sodium hydroxide TS to make exactly 100 mL. Pipet 3 mL
[28981-97-7]
of this solution, add 0.05 mol/L sodium hydroxide TS to
Alprazolam, when dried, contains not less than
98.5z of alprazolam (C17H13ClN4).
under Ultraviolet-visible Spectrophotometry <2.24>. Description Alprazolam occurs as white, crystals or crys-
talline powder.
It is freely soluble in chloroform, soluble in methanol and
of alminoprofen (C13H17NO2)
in ethanol (95), sparingly soluble in acetic anhydride, and
＝ MS × AT/AS × V?/V × 1/C × 27
practically insoluble in water.
MS: Amount (mg) of alminoprofen for assay taken It dissolves in dilute nitric acid.
C: Labeled amount (mg) of alminoprofen (C13H17NO2) in
1 tablet
solution of Alprazolam in ethanol (95) (1 in 200,000) as
Assay Weigh accurately the mass of not less than 20 tablets directed under Ultraviolet-visible Spectrophotometry <2.24>,
of Alminoprofen Tablets, and powder. Weigh accurately and compare the spectrum with the Reference Spectrum:
an amount equivalent to about 60 mg of alminoprofen both spectra exhibit similar intensities of absorption at the
(C13H17NO2), add ethanol (99.5) and shake well, add ethanol same wavelengths.
(99.5) to make exactly 200 mL, and centrifuge. Pipet 2 mL (2) Dissolve 0.05 g of Alprazolam in 0.7 mL of deu-
of the supernatant liquid, add ethanol (99.5) to make exactly terochloroform for nuclear magnetic resonance spectros-
100 mL, and use this solution as the sample solution. Sepa- copy, and determine the spectrum of this solution using
rately, weigh accurately about 30 mg of alminoprofen for tetramethylsilane for nuclear magnetic resonance spectros-
assay, previously dried in vacuum for 1 hour using phospho- copy as an internal reference compound, as directed under
rus (V) oxide as the dessicant, dissolve in ethanol (99.5) to Nuclear Magnetic Resonance Spectroscopy <2.21> (1H): it ex-
make exactly 100 mL. Pipet 2 mL of this solution, add hibits a single signal A at around d 2.6 ppm, doublet signals
ethanol (99.5) to make exactly 100 mL, and use this solution B and C at around d 4.0 ppm and d 5.4 ppm, and a broad
as the standard solution. Determine the absorbances, AT and signal D between d 7.1 ppm and 7.9 ppm. The ratio of inte-
AS, at the wavelength of maximum absorption at about 255 grated intensity of each signal, A:B:C:D, is about 3:1:1:8.
nm of the sample solution and standard solution as directed (3) Perform the test with Alprazolam as directed under
under Ultraviolet-visible Spectrophotometry <2.24>. Flame Coloration Test <1.04> (2): a green color appears.
Amount (mg) of alminoprofen (C13H17NO2) Melting point <2.60> 228 – 2329C
＝ M S × AT / AS × 2
Purity (1) Chloride <1.03>—Dissolve 0.5 g of Alprazolam
MS: Amount (mg) of alminoprofen for assay taken in 10 mL of dilute nitric acid, and add water to make 50 mL.
Perform the test using this solution as the test solution. Pre-
pare the control solution with 0.20 mL of 0.01 mol/L hydro-
ers.
chloric acid VS (not more than 0.014z).
Alprazolam according to Method 4, and perform the test.
(3) Related substances—Dissolve 50 mg of Alprazolam
in 10 mL of methanol, and use this solution as the sample so-
lution. Pipet 1 mL of the sample solution, add methanol to
make exactly 100 mL, then pipet 1 mL of this solution, add
methanol to make exactly 10 mL, and use this solution as the
plate of silica gel with fluorescent indicator for thin-layer
chromatography. Develop with a mixture of acetone,
hexane, ethyl acetate and ethanol (95) (4:2:2:1) to a distance
396 Alprenolol Hydrochloride / Official Monographs JP XVII
of about 10 cm, and air-dry the plate. Examine under ultra- pH <2.54> Dissolve 1.0 g of Alprenolol Hydrochloride in
violet light (main wavelength: 254 nm): the spots other than 10 mL of water: the pH of this solution is between 4.5 and
the principal spot from the sample solution are not more 6.0.
intense than the spot from the standard solution.
Melting point <2.60> 108 – 1129
C
Loss on drying <2.41> Not more than 0.5z (1 g, reduced
pressure not exceeding 0.67 kPa, 609C, 4 hours).
of Alprenolol Hydrochloride in 10 mL of water: the solution
Residue on ignition <2.44> Not more than 0.1z (1 g). is clear and colorless.
Assay Weigh accurately about 0.25 g of Alprazolam,
Alprenolol Hydrochloride according to Method 2, and
previously dried, dissolve in 100 mL of acetic anhydride,
perform the test. Prepare the control solution with 2.0 mL
and titrate <2.50> with 0.1 mol/L perchloric acid VS (poten-
of Standard Lead Solution (not more than 10 ppm).
tiometric titration). Perform a blank determination in the
same manner, and make any necessary correction.
of Alprenolol Hydrochloride according to Method 3, and
Each mL of 0.1 mol/L perchloric acid VS perform the test (not more than 2 ppm).
＝ 15.44 mg of C17H13ClN4 (4) Related substances—Dissolve 0.10 g of Alprenolol
Hydrochloride in 10 mL of ethanol (95), and use this solu-
tion as the sample solution. Pipet 1 mL of the sample solu-
ers.
tion, and add ethanol (95) to make exactly 100 mL. Pipet 2.5
mL of this solution, add ethanol (95) to make exactly 10 mL,
Alprenolol Hydrochloride test with these solutions as directed under Thin-layer Chro-
matography <2.03>. Spot 10 mL each of the sample solution
アルプレノロール塩酸塩
and standard solution on a plate of silica gel for thin-layer
dichloromethane, acetone, acetic acid (100) and water
(60:42:5:3) to a distance of about 10 cm, air-dry the plate,
and then dry at 809C for 30 minutes. After cooling, allow
the plate to stand in iodine vapor for 30 minutes: the spots
C15H23NO2.HCl: 285.81 other than the principal spot and the spot on the starting
(2RS )-1-(2-Allylphenoxy)-3- point from the sample solution are not more intense than the
[(1-methylethyl)amino]propan-2-ol monohydrochloride spot from the standard solution.
[13707-88-5]
Alprenolol Hydrochloride, when dried, contains
not less than 99.0z of alprenolol hydrochloride Residue on ignition <2.44> Not more than 0.1z (1 g).
(C15H23NO2.HCl).
Assay Weigh accurately about 0.5 g of Alprenolol Hydro-
Description Alprenolol Hydrochloride occurs as white, chloride, previously dried, dissolve in 50 mL of a mixture of
crystals or crystalline powder. acetic anhydride and acetic acid (100) (7:3), and titrate <2.50>
It is freely soluble in water, in ethanol (95) and in acetic with 0.1 mol/L perchloric acid VS (potentiometric titration).
acid (100), slightly soluble in acetic anhydride, and practi- Perform a blank determination, and make any necessary
cally insoluble in diethyl ether. correction.
Identification (1) To 2 mL of a solution of Alprenolol Each mL of 0.1 mol/L perchloric acid VS
Hydrochloride (1 in 100) add 0.05 mL of copper (II) sulfate ＝ 28.58 mg of C15H23NO2.HCl
TS and 2 mL of sodium hydroxide TS: a blue-purple color
develops. To this solution add 1 mL of diethyl ether, shake
ers.
well, and allow to stand: a red-purple color develops in the
diethyl ether layer.
(2) Dissolve 0.05 g of Alprenolol Hydrochloride in 5 mL
of water, add 1 to 2 drops of bromine TS, and shake: the Alprostadil
color of the test solution disappears.
(3) Determine the absorption spectrum of a solution of
Prostaglandin E1
Alprenolol Hydrochloride in ethanol (95) (1 in 10,000) as
アルプロスタジル
same wavelengths.
Alprenolol Hydrochloride, previously dried, as directed in
the potassium chloride disk method under Infrared Spectro-
C20H34O5: 354.48
photometry <2.25>, and compare the spectrum with the
7-{(1R,2R,3R)-3-Hydroxy-2-[(1E,3S )-3-
Reference Spectrum: both spectra exhibit similar intensities
hydroxyoct-1-en-1-yl]-5-oxocyclopentyl}heptanoic acid
of absorption at the same wave numbers.
[745-65-3]
(5) A solution of Alprenolol Hydrochloride (1 in 50)
Alprostadil, when dried, contains not less than
JP XVII Official Monographs / Alprostadil 397
97.0z and not more than 103.0z of alprostadil uid chromatography and water (9:1) to make exactly 20 mL.
(C20H34O5). Confirm that the peak area of alprostadil obtained with 5 mL
of this solution is equivalent to 7 to 13z of that obtained
Description Alprostadil occurs as white, crystals or crystal-
with 5 mL of the standard solution.
line powder.
It is freely soluble in ethanol (99.5) and in tetrahydrofu-
ran, slightly soluble in acetonitrile, and practically insoluble
in water.
of alprostadil is not more than 1.5z.
Identification (1) The absorption spectrum of a solution
of Alprostadil in ethanol (99.5) (1 in 100,000) determined as
um, phosphorus (V) oxide, 4 hours).
directed under Ultraviolet-visible Spectrophotometry <2.24>
shows no absorption between 210 nm and 350 nm. Sepa- Assay Weigh accurately about 5 mg each of Alprostadil
rately, to 10 mL of this solution add 1 mL of potassium hy- and Alprostadil RS, previously dried, dissolve in exactly 5
droxide-ethanol TS, allow to stand for 15 minutes, and de- mL of the internal standard solution, add a mixture of aceto-
termine the absorption spectrum in the same way. Compare nitrile for liquid chromatography and water (9:1) to make 50
the spectrum so obtained with the Reference Spectrum or the mL, and use these solutions as the sample solution and
spectrum of a solution of Alprostadil RS prepared in the standard solution, respectively. Perform the test with 5 mL
same manner as the sample solution: both spectra exhibit each of the sample solution and standard solution as directed
similar intensities of absorption at the same wavelengths. under Liquid Chromatography <2.01> according to the fol-
(2) Determine the infrared absorption spectrum of lowing conditions, and calculate the ratios, QT and QS, of
Alprostadil, previously dried, as directed in the potassium the peak area of alprostadil to that of the internal standard.
Amount (mg) of alprostadil (C20H34O5) ＝ MS × QT/QS
trum or the spectrum of previously dried Alprostadil RS: MS: Amount (mg) of Alprostadil RS taken
Internal standard solution—A solution of dimethyl phtha-
same wave numbers.
late in the mixture of acetonitrile for liquid chromatography
D : －53 – －619 (after drying, and water (9:1) (1 in 10,000).
25 mg, tetrahydrofuran, 5 mL, 100 mm). Operating conditions—
length: 196 nm).
Purity Related substances—Dissolve 4 mg of Alprostadil in Column: A stainless steel column 4.6 mm in inside diame-
2 mL of a mixture of acetonitrile for liquid chromatography ter and 15 cm in length, packed with octadecylsilanized silica
and water (9:1), and use this solution as the sample solution. gel for liquid chromatography (5 mm in particle diameter).
Pipet 0.5 mL of the sample solution, and add the mixture of Column temperature: A constant temperature of about
acetonitrile for liquid chromatography and water (9:1) to 409C.
make exactly 10 mL. Pipet 2 mL of this solution, add the Mobile phase: Dissolve 9.07 g of potassium dihydrogen
mixture of acetonitrile for liquid chromatography and water phosphate in water to make 1000 mL. Adjust the pH to 6.3
(9:1) to make exactly 10 mL, and use this solution as the with a solution prepared by dissolving 9.46 g of disodium
standard solution. Perform the test with exactly 5 mL each of hydrogen phosphate in water to make 1000 mL, and dilute to
the sample solution and standard solution as directed under 10 times its volume with water. To 360 mL of this solution
Liquid Chromatography <2.01> according to the following add 110 mL of acetonitrile for liquid chromatography and
conditions, and determine each peak area by the automatic 30 mL of methanol for liquid chromatography.
integration method: the area of the peaks, having the relative Flow rate: Adjust so that the retention time of alprostadil
retention time of about 0.70 and 1.26 to alprostadil, is not is about 10 minutes.
larger than 1/2 times the peak area of alprostadil with the System suitability—
standard solution, the area of the peaks, having the relative System performance: When the procedure is run with 5 mL
retention time of about 0.88 and 1.18 to alprostadil, is not of the standard solution under the above operating condi-
larger than the peak area of alprostadil with the standard so- tions, alprostadil and the internal standard are eluted in this
lution, the area of the peaks other than alprostadil and the order with the resolution between these peaks being not less
peaks mentioned above is not larger than 1/10 times the than 9.
peak area of alprostadil with the standard solution and the System repeatability: When the test is repeated 6 times
total area of the peaks other than alprostadil is not larger with 5 mL of the standard solution under the above operating
than 2 times the peak area of alprostadil with the standard conditions, the relative standard deviation of the ratio of the
solution. peak area of alprostadil to that of the internal standard is
Operating conditions— not more than 1.0z.
Storage—Light-resistant, and at a temperature not
the Assay.
exceeding 59C.
retention time of alprostadil, beginning after the solvent
peak.
System performance: Proceed as directed in the system
Test for required detectability: Measure exactly 2 mL of
the standard solution, add the mixture of acetonitrile for liq-
398 Alprostadil Injection / Official Monographs JP XVII
Amount ( mg) of prostaglandin A1 (C20H32O4), converted
Alprostadil Injection to alprostadil
＝ MS × QT/QS × 1/2 × 1.054
アルプロスタジル注射液
MS: Amount (mg) of prostaglandin A1 taken
Internal standard solution—Dissolve 50 mg of 1-naphthol in
Alprostadil Injection is an emulsion-type injection.
20 mL of ethanol (99.5). To 3 mL of this solution add the
mobile phase to make 100 mL.
than 125.0z of the labeled amount of alprostadil
(C20H34O5: 354.48).
Method of preparation Prepare as directed under Injec- Assay.
tions, with Alprostadil. System suitability—
System performance, and system repeatability: Proceed as
Description Alprostadil Injection occurs as a white emul-
directed in the system suitability in the Assay.
sion and is slightly viscous. It has a distinctive odor.
Test for required detectability: To exactly 1 mL of the
Identification To a quantity of Alprostadil Injection, cor- standard solution add the mobile phase to make exactly 5
responding to 10 mg of Alprostadil, add 2 mL of acetonitrile, mL. Confirm that the peak area of prostaglandin A1 ob-
shake well, and centrifuge. To 3.5 mL of the supernatant tained with 40 mL of this solution is equivalent to 14 to 26z
liquid add 7 mL of diluted phosphoric acid (1 in 1000), and of that obtained with 40 mL of the standard solution.
then run this solution on a column (prepared by filling a 10 (3) Peroxide—Pipet 4 mL of Alprostadil Injection, place
mm inside diameter, 9 mm long chromatography tube with in a glass-stoppered flask, add 15 mL of a mixture of acetic
0.4 g of 70 mm octadecylsilanized silica gel for pretreatment) acid (100) and isooctane (3:2), previously having undergone
prewashed with 10 mL of methanol and then 10 mL of a 30 minute nitrogen substitution, and dissolve with gentle
water. Wash the column with 10 mL of water and then 20 shaking. To this solution add 0.5 mL of saturated potassium
mL of petroleum ether, followed by elution with 2.5 mL of a iodide TS, replace the inside of the vessel with nitrogen, and
mixture of methanol and water (4:1). Remove the solvent shake for exactly 5 minutes. Then, add 0.5 mL of starch TS,
from the effluent under reduced pressure, dissolve the shake vigorously, add 15 mL of water, and shake vigorously.
residue in 100 mL of ethyl acetate, and use this solution as Under a stream of nitrogen, titrate <2.50> with 0.01 mol/L
the sample solution. Separately, dissolve 1 mg of Alprostadil sodium thiosulfate VS until the color of the solution disap-
RS in 10 mL of ethyl acetate, and use this solution as the pears. Separately, perform a blank determination using 4
standard solution. Perform the test with these solutions as mL of water, and make any necessary correction. Calculate
directed under Thin-layer Chromatography <2.03>. Spot the the amount of peroxides using the following equation: not
entire volume of the sample solution and 100 mL of the more than 0.5 meq/L.
standard solution on a plate of silica gel for thin-layer chro-
Amount (meq/L) of peroxides ＝ V × 2.5
matography. Then, develop the plate with a mixture of ethyl
acetate, ethanol (99.5) and acetic acid (100) (100:5:1) to a V: Amount (mL) of 0.01 mol/L sodium thiosulfate VS
distance of about 10 cm, and air-dry the plate. Spray evenly consumed
a solution of phosphomolybdic acid n-hydrate in ethanol
(4) Free fatty acids—Pipet 3 mL of Alprostadil Injec-
(99.5) (1 in 10) on the plate, and heat at 1009C for 5 minutes:
tion, add exactly 15 mL of a mixture of 2-propanol, heptane
the color of the spot obtained from the standard solution
and 0.5 mol/L sulfuric acid TS (40:10:1), and shake for 1
and the spot corresponding to that location obtained from
minute. After leaving for 10 minutes, add exactly 9 mL of
the sample solution is dark blue.
heptane and exactly 9 mL of water, shake the test tube by in-
pH Being specified separately when the drug is granted ap- verting 10 times, leave for 15 minutes, and pipet 9 mL of the
proval based on the Law. supernatant liquid. To this solution, add 3 mL of a solution
prepared by combining 1 volume of Nile blue solution (1 in
Purity (1) Heavy metals <1.07>—Proceed with 4.0 mL of
5000) washed 5 times with heptane and 9 volumes of ethanol
Alprostadil Injection according to Method 2, and perform
(99.5), and use this solution as the sample solution. Titrate
<2.50> this solution with 0.02 mol/L sodium hydroxide VS
under a stream of nitrogen. Separately, dissolve 5.65 g of
(2) Prostaglandin A1—Use the sample solution obtained
oleic acid in heptane to make exactly 200 mL, and use this
in the Assay as the sample solution. Separately, weigh accu-
solution as the standard solution. Pipet 25 mL of the stand-
rately about 10 mg of prostaglandin A1, previously dried for
ard solution, add 2 drops of phenolphthalein TS, titrate
4 hours in a desiccator (in vacuum, phosphorus (V) oxide),
<2.50> with 0.1 mol/L potassium hydroxide-ethanol VS until
and dissolve in ethanol (99.5) to make exactly 100 mL. Pipet
a light red color develops, and determine the correction fac-
2.5 mL of this solution, and add the mobile phase to make
tor f. Pipet 30 mL of the standard solution and add heptane
exactly 50 mL. Pipet 1 mL of this solution, add exactly 1 mL
of the internal standard solution, and use this solution as the
exactly 15 mL of a mixture of 2-propanol, heptane and 0.5
standard solution. Perform the test with 40 mL each of the
mol/L sulfuric acid TS (40:10:1), and shake for 1 minute.
sample solution and standard solution as directed under Liq-
After leaving for 10 minutes, add exactly 6 mL of heptane
uid Chromatography <2.01> according to the following con-
and exactly 12 mL of water, shake the test tube by inverting
ditions, calculate the ratios, QT and QS, of the peak area of
10 times, and then titrate <2.50> in the same manner as for
prostaglandin A1 to that of the internal standard, and calcu-
the sample solution. Determine the volume (mL), VT and VS,
late the amount of prostaglandin A1 converted to alprostadil
of 0.02 mol/L sodium hydroxide VS consumed by the sam-
using the following equation: not more than 3.0 mg per a
ple and standard solutions: the amount of free fatty acid is
volume, equivalent to 5 mg of alprostadil (C20H34O5).
not more than 12.0 meq/L.
Amount (meq/L) of free fatty acids ＝ VT/VS × f × 15
JP XVII Official Monographs / Alprostadil Injection 399
Bacterial endotoxins <4.01> Less than 10 EU/mL. solution, and routing valve for 2 high pressure flow paths.
Pretreatment column: A stainless steel column 4 mm in
inside diameter and 2.5 cm in length, packed with octadecyl-
Foreign insoluble matter <6.06> Perform the test according silanized silica gel for liquid chromatography (5 mm in parti-
to Method 1: it meets the requirement. cle diameter).
Pretreatment column wash solution: Ethanol (99.5).
Sterility <4.06> Perform the test according to the Mem-
Flow rate of wash solution: A constant flow rate of about
brane filter method: it meets the requirement. However,
2.0 mL per minute.
use the sample solution consisting of equal volume of
Flow path operating conditions: Change the flow path
Alprostadil Injection and a solution prepared by adding
operating conditions at the times shown in the table below
water to 0.1 g of polysorbate 80 to make 100 mL.
using the valves shown in the figure.
Particle diameter Being specified separately when the drug
is granted approval based on the Law.
Time of switchover (minutes)
Assay Measure exactly a volume of Alprostadil Injection
corresponding to 5 mg of alprostadil (C20H34O5), add exactly Valve 0 9.0 9.1 *1) *2)
1 mL of the internal standard solution, shake, and use this
RVA 0 0 1 0 0
about 5 mg of Alprostadil RS, previously dried in a desicca- RVB 0 1 1 1 0
tor (in vacuum, phosphorus (V) oxide) for 4 hours, dissolve
in ethanol (99.5) to make exactly 50 mL, and use this solu- *1) After the internal standard has completely eluted
tion as standard stock solution. Pipet 2.5 mL of the standard *2) 0.1 minutes after *1)
stock solution, add the mobile phase to make exactly 50 mL,
pipet 1 mL, add exactly 1 mL of the internal standard solu- System suitability—
tion, and use this solution as the standard solution. Perform System performance: Dissolve 10 mg of prostaglandin A1,
the test with 40 mL each of the sample solution and standard previously dried in a desiccator (in vacuum, phosphorus (V)
solution as directed under Liquid Chromatography <2.01> oxide) for 4 hours, in ethanol (99.5) to make 100 mL. To 2.5
according to the following conditions using an apparatus mL of this solution add 2.5 mL of the standard stock solu-
equipped with an automatic pretreatment device (using a tion, and add the mobile phase to make 50 mL. To 1 mL of
postcolumn reaction), and calculate the ratios, QT and QS, of this solution add 1 mL of the internal standard solution,
the peak area of alprostadil to that of the internal standard. shake, and perform the test under the above conditions with
40 mL of the solution. Alprostadil, prostaglandin A1 and the
Amount ( mg) of alprostadil (C20H34O5) ＝ MS × QT/QS internal standard are eluted in this order, and the resolution
MS: Amount (mg) of Alprostadil RS taken between the peaks of alprostadil and prostaglandin A1 is not
less than 10, and that between prostaglandin A1 and the in-
Internal standard solution—Dissolve 50 mg of 1-naphthol in ternal standard is not less than 2.0.
20 mL of ethanol (99.5). To 3 mL of this solution add the System repeatability: When the test is repeated 6 times
mobile phase to make 100 mL. with 40 mL of the standard solution under the above condi-
Operating conditions— tions, the relative standard deviation of the ratio of the peak
Equipment: Liquid chromatograph consisting of 2 pumps area of alprostadil to that of the internal standard is not
for pumping the mobile phase and the reaction reagent, an more than 2.0z.
automatic pretreatment device, column, reaction coil, detec-
tor, and recording apparatus. Use a reaction coil that is Containers and storage Containers—Hermetic containers.
maintained at a constant temperature. Storage—Light-resistant, not exceeding 59C, avoiding
Detector: An ultraviolet absorption photometer (wave- freezing.
length: 278 nm).
609 C.
Reaction coil: Polytetrafluoroethylene tube 0.5 mm in
inside diameter and 10 m in length.
Mobile phase: Dissolve 9.07 g of potassium dihydrogen
phosphate in water to make 1000 mL and adjust the pH to
6.3 by adding a solution prepared by dissolving 9.46 g of
disodium hydrogen phosphate in water to make 1000 mL. A: RVA valve
To 1 volume of this solution add 9 volumes of water. To 3 B: RVB valve
volumes of this solution add 1 volume of acetonitrile for C: Sample injector
liquid chromatography. D: Mobile phase
Reaction reagent: Potassium hydroxide TS. E: Column for pressure correction
Reaction temperature: A constant temperature of about F: Column
609 C. G: Pretreatment column
Mobile phase flow rate: Adjust so that the retention time H: Wash solution
of alprostadil is about 7 minutes. I: Drain
Reaction reagent flow rate: 0.5 mL per minute. J: Pump
Automatic pretreatment device: Composed of a pretreat-
ment column, pump for pumping pretreatment column wash Figure Components of automatic pretreatment system
400 Alprostadil Alfadex / Official Monographs JP XVII
of water: the pH of this solution is between 4.0 and 5.0.
Alprostadil Alfadex Purity (1) Clarity and color of solution—Dissolve 1.0 g
of Alprostadil Alfadex in 10 mL of water: the solution is col-
Prostaglandin E1 a-Cyclodextrin Clathrate orless. Perform the test with this solution as directed under
Compound Ultraviolet-visible Spectrophotometry within 30 minutes
after preparation of the solution: the absorbance at 450 nm
アルプロスタジルアルファデクス is not larger than 0.10.
(2) Prostaglandin A1—Dissolve 0.10 g of Alprostadil
Alfadex in 5 mL of water, add exactly 5 mL of the internal
standard solution and ethanol (95) to make 15 mL, and use
this solution as the sample solution. Separately, dissolve 1.5
mg of prostaglandin A1 in ethanol (95) to make exactly 100
mL. Pipet 3 mL of this solution, add exactly 5 mL of the in-
ternal standard solution, 2 mL of ethanol (95) and water to
C20H34O5.xC36H60O30 make 15 mL, and use this solution as the standard solution.
7-{(1R,2R,3R)-3-Hydroxy-2-[(1E,3S )-3-hydroxyoct-1- Perform the test with 10 mL each of the sample solution and
en-1-yl]-5-oxocyclopentyl}heptanoic acid—a-cyclodextrin standard solution as directed under Liquid Chromatography
[55648-20-9] <2.01> according to the operating conditions described in the
Assay, and calculate the ratios, QT and QS, of the peak area
Alprostadil Alfadex is a a-cyclodextrin clathrate of prostaglandin A1 to that of the internal standard: QT is
compound of alprostadil. not larger than QS.
It contains not less than 2.8z and not more than Internal standard solution—A solution of propyl parahy-
3.2z of alprostadil (C20H34O5: 354.48), calculated on droxybenzoate in dilute ethanol (1 in 15,000).
the anhydrous basis. (3) Related substances—Dissolve 0.10 g of Alprostadil
Alfadex in 3 mL of water, add exactly 3 mL of ethyl acetate,
Description Alprostadil Alfadex occurs as a white powder. shake, centrifuge, and use the supernatant liquid obtained
It is freely soluble in water, and practically insoluble in as the sample solution. Separately, dissolve 1.0 mg of
ethanol (95), in ethyl acetate and in diethyl ether. prostaglandin A1 in ethyl acetate to make exactly 100 mL,
It is hygroscopic. and use this solution as the standard solution. Perform the
Identification (1) Dissolve 0.02 g of Alprostadil Alfadex test with these solutions as directed under Thin-layer Chro-
in 5 mL of water, add 5 mL of ethyl acetate, shake, and matography <2.03>. Spot 10 mL each of the sample solution
centrifuge. Use the supernatant liquid as the sample solution and standard solution on a plate of silica gel for thin-layer
(1). Separately, to 0.02 g of Alprostadil Alfadex add 5 mL of chromatography. Develop the plate with a mixture of ethyl
ethyl acetate, shake, and centrifuge. Use the supernatant acetate, hexane and acetic acid (100) (10:2:1) to a distance of
liquid as the sample solution (2). Evaporate the solvent from about 10 cm, and air-dry the plate. Spray evenly a solution
these solutions under reduced pressure, add 2 mL of sulfuric of phosphomolybdic acid n-hydrate in ethanol (95) (1 in 4)
acid to the residue, and shake for 5 minutes: the liquid on the plate, and heat at 1009 C for 5 minutes: the spots
obtained from the sample solution (1) shows an orange- other than the principal spot from the sample solution, and
yellow color, while the liquid obtained from the sample solu- the spots other than the spot corresponding to the spot from
tion (2) does not show that color. the standard solution are all not more intense than the spot
(2) Dissolve 0.02 g of Alprostadil Alfadex in 5 mL of from the standard solution.
water, add 5 mL of ethyl acetate, shake, centrifuge, and Water <2.48> Not more than 6.0z (0.2 g, direct titration).
evaporate the solvent from the supernatant liquid under
reduced pressure. Dissolve the residue in 2 mL of ethanol Assay Weigh accurately about 0.1 g of Alprostadil
(95), add 5 mL of 1,3-dinitrobenzene TS, then add 5 mL of a Alfadex, dissolve in 5 mL of water, add exactly 5 mL of the
solution of potassium hydroxide in ethanol (95) (17 in 100) internal standard solution and water to make 15 mL, and use
under ice-cooling, and allow to stand for 20 minutes in a this solution as the sample solution. Separately, weigh
dark place under ice-cooling: a purple color develops. accurately about 3 mg of Alprostadil RS, dissolve in 5 mL of
(3) Dissolve 0.05 g of Alprostadil Alfadex in 1 mL of ethanol (95), add exactly 5 mL of the internal standard solu-
iodine TS, by heating on a water bath, and allow to stand: a tion and water to make 15 mL, and use this solution as the
dark blue precipitate is formed. standard solution. Perform the test with 10 mL each of the
(4) Determine the absorption spectrum of a solution of sample solution and standard solution as directed under
Alprostadil Alfadex in dilute ethanol (3 in 10,000) as Liquid Chromatography <2.01> according to the following
directed under Ultraviolet-visible Spectrophotometry <2.24>: conditions, and calculate the ratios, QT and QS, of the peak
it exhibits no absorption between 220 nm and 400 nm. area of alprostadil to that of the internal standard.
Separately, to 10 mL of the solution add 1 mL of potassium Amount (mg) of alprostadil (C20H34O5)
hydroxide-ethanol TS, allow to stand for 15 minutes, and ＝ M S × Q T / QS
determine the absorption spectrum as directed under Ultravi-
olet-visible Spectrophotometry <2.24>, and compare the MS: Amount (mg) of Alprostadil RS taken
spectrum with the Reference Spectrum: both spectra exhibit Internal standard solution—A solution of propyl parahy-
similar intensities of absorption at the same wavelengths. droxybenzoate in dilute ethanol (1 in 15,000).
Optical rotation <2.49> [a]20 Operating conditions—
D : ＋126 – ＋1389(0.1 g calcu-
lated on the anhydrous basis, dilute ethanol, 20 mL, 100 Detector: An ultraviolet absorption photometer (wave-
mm). length: 205 nm).
Column: A stainless steel column about 5 mm in inside
pH <2.54> Dissolve 0.10 g of Alprostadil Alfadex in 20 mL diameter and about 15 cm in length, packed with octadecyl-
JP XVII Official Monographs / Dried Aluminum Hydroxide Gel 401
silanized silica gel for liquid chromatography (5 mm in parti-

cle diameter). Dried Aluminum Hydroxide Gel
259 C. 乾燥水酸化アルミニウムゲル
drogenphosphate and acetonitrile (3:2).
Dried Aluminum Hydroxide Gel contains not less
Flow rate: Adjust so that the retention time of alprostadil
than 50.0z of aluminum oxide (Al2O3: 101.96).
is about 6 minutes.
Selection of column: Dissolve about 0.1 g of Alprostadil Description Dried Aluminum Hydroxide Gel occurs as a
Alfadex in 5 mL of water, add 5 mL of a solution of white, amorphous powder. It is odorless and tasteless.
prostaglandin A1 in ethanol (95) (3 in 200,000) and 5 mL of It is practically insoluble in water, in ethanol (95) and in
the internal standard solution. Proceed with 10 mL of this diethyl ether.
solution under the above operating conditions, and calculate Most of it dissolves in dilute hydrochloric acid and in
the resolution. Use a column giving elution of alprostadil, sodium hydroxide TS.
the internal standard and prostaglandin A1 in this order and
Identification To 0.2 g of Dried Aluminum Hydroxide Gel
complete separation of these peaks.
add 20 mL of dilute hydrochloric acid, warm, and centri-
Containers and storage Containers—Tight containers. fuge: the supernatant liquid responds to the Qualitative Tests
Storage—Light-resistant, at a temperature not exceeding <1.09> for aluminum salt.
59C.
Purity (1) Acidity or alkalinity—To 1.0 g of Dried Alu-
minum Hydroxide Gel add 25 mL of water, shake well, and
centrifuge: the supernatant liquid is neutral.
Alum Solution (2) Chloride <1.03>—To 1.0 g of Dried Aluminum Hy-
droxide Gel add 30 mL of dilute nitric acid, heat gently to
ミョウバン水
boil while shaking, cool, add water to make 100 mL, and
centrifuge. To 5 mL of the supernatant liquid add 6 mL of
Alum Solution contains not less than 0.27 w/vz dilute nitric acid and water to make 50 mL. Perform the test
and not more than 0.33 w/vz of aluminum potassium using this solution as the test solution. Prepare the control
sulfate Hydrate [AlK(SO4)2.12H2O: 474.39]. solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS
(not more than 0.284z).
(3) Sulfate <1.14>—To 1.0 g of Dried Aluminum Hy-
Aluminum Potassium Sulfate Hydrate 3g droxide Gel add 15 mL of dilute hydrochloric acid, heat
Mentha Water 50 mL gently to boil while shaking, cool, add water to make 250
Water, Purified Water or Purified mL, and centrifuge. To 25 mL of the supernatant liquid add
Water in Containers a sufficient quantity 1 mL of dilute hydrochloric acid and water to make 50 mL.
To make 1000 mL Perform the test using this solution as the test solution. Pre-
pare the control solution with 1.0 mL of 0.005 mol/L sulfu-
Dissolve and mix the above ingredients. ric acid VS (not more than 0.480z).
Description Alum Solution is a clear, colorless liquid. It (4) Nitrate—To 0.10 g of Dried Aluminum Hydroxide
has the odor of the mentha oil and an astringent taste. Gel add 5 mL of water, then carefully add 5 mL of sulfuric
acid, shake well to dissolve, and cool. Superimpose the solu-
Identification (1) To 5 mL of Alum Solution add 3 mL of tion on 2 mL of iron (II) sulfate TS: no brown-colored ring
ammonium chloride TS and 1 mL of ammonia TS: a white, is produced at the zone of contact.
gelatinous precipitate is produced, which changes to red (5) Heavy metals <1.07>—Dissolve 2.0 g of Dried Alumi-
upon the addition of 5 drops of alizarin red S TS (aluminum num Hydroxide Gel in 10 mL of dilute hydrochloric acid by
sulfate). heating, filter if necessary, and add water to make 50 mL.
(2) Place 100 mL of Alum Solution in an evaporating Perform the test with this solution as the test solution.
dish, evaporate on a water bath to dryness, and dissolve the Prepare the control solution as follows: evaporate 10 mL of
residue in 5 mL of water: the solution responds to the dilute hydrochloric acid to dryness, and add 2.0 mL of
Qualitative Tests <1.09> for potassium salt. Standard Lead Solution, 2 mL of dilute acetic acid and water
(3) Alum Solution responds to the Qualitative Tests to make 50 mL (not more than 10 ppm).
<1.09> (1) and (2) for sulfate. (6) Arsenic <1.11>—To 0.8 g of Dried Aluminum Hy-
Assay Pipet 50 mL of Alum Solution, add exactly 30 mL droxide Gel add 10 mL of dilute sulfuric acid, heat gently to
of 0.02 mol/L disodium dihydrogen ethylenediamine tetra- boil while shaking, cool, and filter. Take 5 mL of the fil-
acetate VS, and further add 20 mL of acetic acid-ammonium trate, use this solution as the test solution, and perform the
acetate buffer solution (pH 4.8). Boil for 5 minutes, cool, test (not more than 5 ppm).
add 55 mL of ethanol (95), and titrate <2.50> with 0.02 Acid-consuming capacity Weigh accurately about 0.2 g of
mol/L zinc acetate VS (indicator: 2 mL of dithizone TS), Dried Aluminum Hydroxide Gel, and transfer to a glass-
until the color of the solution changes from light dark green stoppered flask. Add exactly 100 mL of 0.1 mol/L hydro-
to light red. Perform a blank determination. chloric acid VS, stopper the flask, shake at 37 ± 29C for
Each mL of 0.02 mol/L disodium dihydrogen ethylene- 1 hour, and filter. Measure exactly 50 mL of the filtrate, and
diamine tetraacetate VS titrate <2.50> while thoroughly stirring, the excess hydrochlo-
＝ 9.488 mg of AlK(SO4)2.12H2O ric acid with 0.1 mol/L sodium hydroxide VS until the pH of
the solution becomes to 3.5. The volume of 0.1 mol/L hy-
Containers and storage Containers—Tight containers. drochloric acid VS consumed is not less than 250 mL per g of
Dried Aluminum Hydroxide Gel.
402 Dried Aluminum Hydroxide Gel Fine Granules / Official Monographs JP XVII
Assay Weigh accurately about 2 g of Dried Aluminum Hy- Identification (1) Heat 3 g of Aluminum Monostearate
droxide Gel, add 15 mL of hydrochloric acid, heat on a with 30 mL of hydrochloric acid in a water bath with occa-
water bath with shaking for 30 minutes, cool, and add water sional shaking for 10 minutes. After cooling, shake the mix-
to make exactly 500 mL. Pipet 20 mL of this solution, add ture vigorously with 50 mL of water and 30 mL of diethyl
exactly 30 mL of 0.05 mol/L disodium dihydrogen ethylene- ether for 3 minutes, and allow to stand. To the separated
diamine tetraacetate VS and 20 mL of acetic acid (31)-am- aqueous layer add sodium hydroxide TS until the solution
monium acetate buffer solution (pH 4.8), boil for 5 minutes, becomes slightly turbid, and filter: the filtrate responds to
and cool. Add 55 mL of ethanol (95), and titrate <2.50> with the Qualitative Tests <1.09> for aluminum salt.
0.05 mol/L zinc acetate VS until the color of the solution (2) Wash the diethyl ether layer separated in (1) with two
changes from light dark green to light red. (indicator: 2 mL 20-mL portions of water, and evaporate the diethyl ether
of dithizone TS). Perform a blank determination, and make layer on a water bath: the residue melts <1.13> at above
any necessary correction. 549C.
Each mL of 0.05 mol/L disodium dihydrogen Acid value for fatty acid <1.13> 193 – 210. Weigh accu-
ethylenediamine tetraacetate VS rately about 1 g of fatty acid obtained in the Identification
＝ 2.549 mg of Al2O3 (2), transfer a 250-mL glass-stoppered flask, add 100 mL of
a mixture of diethyl ether and ethanol (95) (2:1), warm to
dissolve, add several drops of phenolphthalein TS, and
proceed as directed under Acid Value.
Dried Aluminum Hydroxide Gel Purity (1) Free fatty acid—Mix 1.0 g of Aluminum
Monostearate with about 50 mL of a mixture of neutralized
Fine Granules ethanol and diethyl ether (1:1), filter through dry filter
paper, wash the vessel and the filter paper with a small
乾燥水酸化アルミニウムゲル細粒
amount of a mixture of neutralized ethanol and diethyl ether
(1:1), combine the filtrate and the washings, and add 2.1 mL
Dried Aluminum Hydroxide Gel Fine Granules con- of 0.1 mol/L potassium hydroxide VS: a red color develops.
tain not less than 47.0z of aluminum oxide (Al2O3: (2) Water-soluble salts—Heat 2.0 g of Aluminum
101.96). Monostearate with 80 mL of water in a loosely stoppered
conical flask on a water bath for 30 minutes with occasional
shaking. After cooling, filter through dry filter paper, wash
ules, with Dried Aluminum Hydroxide Gel.
the residue with a small amount of water, combine the wash-
Identification To 0.2 g of Dried Aluminum Hydroxide Gel ings with the filtrate, add water to make 100 mL, evaporate
Fine Granules add 20 mL of dilute hydrochloric acid, warm 50 mL of this solution on a water bath, and heat strongly at
and centrifuge: the supernatant liquid responds to the Quali- 6009C: the mass of the residue is not more than 10.0 mg.
tative Tests <1.09> for aluminum salt. (3) Heavy metals <1.07>—Heat 1.0 g of Aluminum
Monostearate over a small flame with caution at the begin-
Acid-consuming capacity Proceed as directed for Acid-
ning, and continue the heating, gradually raising the temper-
consuming capacity under Dried Aluminum Hydroxide Gel:
ature, to ash. After cooling, add 10 mL of diluted hydro-
the volume of 0.1 mol/L hydrochloric acid VS consumed is
chloric acid (1 in 2), evaporate on a water bath, and boil the
not less than 235 mL per g of Dried Aluminum Hydroxide
residue with 20 mL of water for 1 minute. Cool, filter, wash
Gel Fine Granules.
the residue with water, combine the filtrate and the wash-
Assay Proceed as directed in the Assay under Dried Alumi- ings, and add 2 mL of dilute acetic acid and water to make
num Hydroxide Gel. 50 mL. Perform the test using this solution as the test solu-
tion. Evaporate 10 mL of diluted hydrochloric acid (1 in 2)
on a water bath to dryness, add 2 mL of dilute acetic acid
ethylenediamine tetraacetate VS
and 5.0 mL of Standard Lead Solution, dilute with water to
＝ 2.549 mg of Al2O3
make 50 mL, and use this solution as the control solution
Containers and storage Containers—Tight containers. (not more than 50 ppm).
(4) Arsenic <1.11>—Mix 1.0 g of Aluminum
Monostearate with 2 g of magnesium nitrate hexahydrate,
Aluminum Monostearate ignite over a small flame, moisten the residue after cooling
with 0.5 mL of nitric acid, and heat. Heat again the residue
モノステアリン酸アルミニウム with 10 mL of dilute sulfuric acid until white fumes evolve,
add water to make 5 mL, and perform the test with this solu-
tion as the test solution (not more than 2 ppm).
Aluminum Monostearate is mainly aluminum com-
pounds of stearic acid (C18H36O2: 284.48) and palmitic Loss on drying <2.41> Not more than 3.0z (1 g, 1059C,
acid (C16H32O2: 256.42). 3 hours).
Aluminum Monostearate, when dried, contains not
Assay Weigh accurately about 1 g of Aluminum
less than 7.2z and not more than 8.9z of aluminum
Monostearate, previously dried, ignite gently to ash, and
(Al: 26.98).
cool. Add dropwise 0.5 mL of nitric acid, evaporate on a
Description Aluminum Monostearate occurs as a white to water bath by heating, and then heat strongly between 9009
C
yellowish white powder. It is odorless or has a faint, charac- and 11009C to a constant mass. After cooling, weigh rapidly
teristic odor. the ignited residue, and designate the mass as aluminum
It is practically insoluble in water, in ethanol (95) and in oxide (Al2O3: 101.96).
diethyl ether.
JP XVII Official Monographs / Aluminum Potassium Sulfate Hydrate 403
Amount (mg) of aluminum (Al) green to light red. Perform a blank determination.
＝ amount (mg) of aluminum oxide (Al2O3) × 0.529
Containers and storage Containers—Well-closed contain- ethylenediamine tetraacetate VS
ers. ＝ 12.91 mg of AlK (SO4)2
Dried Aluminum Potassium Sulfate

Burnt Alum Aluminum Potassium Sulfate
Hydrate
乾燥硫酸アルミニウムカリウム
Alum
AlK(SO4)2: 258.21
硫酸アルミニウムカリウム水和物
Dried Aluminum Potassium Sulfate, when dried,
contains not less than 98.0z of aluminum potassium AlK(SO4)2.12H2O: 474.39
sulfate [AlK(SO4)2].
Aluminum Potassium Sulfate Hydrate contains not
Description Dried Aluminum Potassium Sulfate occurs as
less than 99.5z of aluminum potassium sulfate hy-
white masses or white powder. It is odorless. It has a slightly
drate [AlK(SO4)2.12H2O].
sweet, astringent taste.
It is freely soluble in hot water and practically insoluble in Description Aluminum Potassium Sulfate Hydrate occurs
ethanol (95). as colorless or white, crystals or powder. It is odorless. It has
It dissolves slowly in water. a slightly sweet, strongly astringent taste.
It is freely soluble in water, and practically insoluble in
Identification A solution of Dried Aluminum Potassium
ethanol (95) and in diethyl ether.
Sulfate (1 in 20) responds to the Qualitative Tests <1.09> for
A solution of Aluminum Potassium Sulfate Hydrate (1 in
aluminum salt, to the Qualitative Tests <1.09> (1), (3) and (4)
20) is acid.
for potassium salt, and to the Qualitative Tests <1.09> (1)
and (3) for sulfate. Identification A solution of Aluminum Potassium Sulfate
Hydrate (1 in 10) responds to the Qualitative Tests <1.09> for
Purity (1) Water-insoluble substances—To 2.0 g of Dried
aluminum salt, to the Qualitative Tests <1.09> (1), (3) and (4)
Aluminum Potassium Sulfate add 40 mL of water, shake
for potassium salt, and to the Qualitative Tests <1.09> (1)
frequently, and allow to stand for 48 hours. Collect the
and (3) for sulfate.
insoluble residue on a glass filter (G4), wash with 50 mL of
water, and dry at 1059C for 2 hours: the mass of the residue Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
is not more than 50 mg. Aluminum Potassium Sulfate Hydrate according to Method
(2) Heavy metals <1.07>—Dissolve 0.5 g of Dried Alumi- 1, and perform the test. Prepare the control solution with 2.0
num Potassium Sulfate in 45 mL of water, and filter, if mL of Standard Lead Solution (not more than 20 ppm).
necessary. Add 2 mL of dilute acetic acid and water to make (2) Iron <1.10>—Prepare the test solution with 1.0 g of
50 mL, and perform the test using this solution as the test so- Aluminum Potassium Sulfate Hydrate according to Method
lution. Prepare the control solution with 2.0 mL of Standard 1, and perform the test according to Method A. Prepare the
Lead Solution, 2 mL of dilute acetic acid and water to make control solution with 2.0 mL of Standard Iron Solution (not
50 mL (not more than 40 ppm). more than 20 ppm).
(3) Iron <1.10>—Prepare the test solution with 0.54 g of (3) Arsenic <1.11>—Prepare the test solution with 0.6 g
Dried Aluminum Potassium Sulfate according to Method 1, of Aluminum Potassium Sulfate Hydrate, according to
and perform the test according to Method A. Prepare the Method 1, and perform the test (not more than 3.3 ppm).
control solution with 2.0 mL of Standard Iron Solution (not
Assay Weigh accurately about 4.5 g of Aluminum Potas-
more than 37 ppm).
sium Sulfate Hydrate, and dissolve in water to make exactly
200 mL. Take exactly 20 mL of this solution, and add ex-
of Dried Aluminum Potassium Sulfate, according to Method
actly 30 mL of 0.05 mol/L disodium dihydrogen ethylene-
1, and perform the test (not more than 5 ppm).
diamine tetraacetate VS and 20 mL of acetic acid-ammo-
Loss on drying <2.41> Not more than 15.0z (2 g, 2009
C, nium acetate buffer solution (pH 4.8), boil for 5 minutes,
4 hours). and cool. Add 55 mL of ethanol (95), and titrate <2.50> with
0.05 mol/L zinc acetate VS (indicator: 2 mL of dithizone
Assay Weigh accurately about 1.2 g of Dried Aluminum
TS), until the color of the solution changes from light dark
Potassium Sulfate, previously dried, add 80 mL of water,
green to light red. Perform a blank determination.
and heat on a water bath with occasional shaking for 20
minutes. Cool, add water to make exactly 100 mL, and Each mL of 0.05 mol/L disodium dihydorgen
filter, if necessary. Discard the first 30 mL of the filtrate, ethylenediamine tetetraacetate VS
take exactly the subsequent 20 mL of the filtrate, and add ＝ 23.72 mg of AlK(SO4)2.12H2O
exactly 30 mL of 0.05 mol/L disodium dihydrogen ethylene-
diamine tetraacetate VS and 20 mL of acetic acid-ammo-
nium acetate buffer solution (pH 4.8), boil for 5 minutes,
and cool. Add 55 mL of ethanol (95), and titrate <2.50> with
0.05 mol/L zinc acetate VS (indicator: 2 mL of dithizone
TS), until the color of the solution changes from light dark
404 Natural Aluminum Silicate / Official Monographs JP XVII
Natural Aluminum Silicate

天然ケイ酸アルミニウム
Description Natural Aluminum Silicate occurs as a white

or slightly colored powder. It is odorless and tasteless.
It is practically insoluble in water, in ethanol (95) and in
diethyl ether.
Natural Aluminum Silicate (1 g) dissolves when heated in
20 mL of a solution of sodium hydroxide (1 in 5), with some
decomposition, leaving a large amount of insoluble sub-
stance.
Identification (1) To 0.5 g of Natural Aluminum Silicate
add 3 mL of diluted sulfuric acid (1 in 3), heat until white
fumes evolve, cool, add 20 mL of water, and filter. Render
the filtrate slightly acid with ammonia TS: the solution re-
sponds to the Qualitative Tests <1.09> for aluminum salt.
(2) Prepare a bead by fusing ammonium sodium
hydrogenphosphate tetrahydrate on a platinum loop. Place
the bead in contact with Natural Aluminum Silicate, and
fuse again: an infusible material appears in the bead,
producing, upon cooling, an opaque bead with a web-like
structure.
Purity (1) Acidity or alkalinity—Shake 5.0 g of Natural
Aluminum Silicate with 100 mL of water, and centrifuge: the
supernatant liquid so obtained is neutral.
(2) Chloride <1.03>—To 5.0 g of Natural Aluminum Sili-
cate add 100 mL of water, boil gently for 15 minutes while
shaking, then cool, add water to restore the original volume,
combined extracts on a water bath to 5 mL. Use this solution
and centrifuge. To 10 mL of the supernatant liquid add 6
as the test solution, and perform the test (not more than 2
mL of dilute nitric acid, dilute to 50 mL with water, and per-
ppm).
form the test using this solution as the test solution. Prepare
(6) Soluble salts—Evaporate 50 mL of the supernatant
the control solution with 0.30 mL of 0.01 mol/L hydrochlo-
liquid obtained in (1) on a water bath to dryness, and ignite
ric acid VS (not more than 0.021z).
the residue at 7009C for 2 hours: the mass of the ignited
(3) Sulfate <1.14>—To the residue obtained in (6) add 3
residue is not more than 40 mg.
mL of dilute hydrochloric acid, heat on a water bath for 10
(7) Fluoride—(i) Apparatus: Use a hard glass appa-
minutes, dilute to 50 mL with water, and filter. To 2.0 mL of
ratus as illustrated in the figure. Ground-glass joints may be
the filtrate add 1 mL of dilute hydrochloric acid and water to
used.
make 50 mL. Perform the test using this solution as the test
(ii) Procedure: Transfer 5.0 g of Natural Aluminum Sili-
solution. Prepare the control solution with 1.0 mL of 0.005
cate to the distilling flask A with the aid of 20 mL of water,
add about 1 g of glass wool and 50 mL of diluted purified
(4) Heavy metals <1.07>—To 1.5 g of Natural Aluminum
sulfuric acid (1 in 2), and connect A to the distillation appa-
Silicate add 50 mL of water and 5 mL of hydrochloric acid,
ratus, previously washed with steam streamed through the
boil gently for 20 minutes while shaking, then cool, centri-
steam introducing tube E. Connect the condenser C with
fuge, remove the supernatant liquid, wash the residue with
the receiver D containing 10 mL of 0.01 mol/L sodium
two 10-mL portions of water, centrifuging each time, com-
hydroxide VS and 10 mL of water so that the lower end of C
bine these washings with the filtrate, and add ammonia solu-
is immersed in the solution. Heat A gradually until the tem-
tion (28) dropwise, until a precipitate just appears. Add
perature of the solution in A reaches 1309C, then open the
dropwise dilute hydrochloric acid with vigorous shaking and
rubber tube F, close the rubber tube G, boil water in the
redissolve the precipitate. Heat the mixture with 0.45 g of
steam generator B vigorously, and introduce the generated
hydroxylammonium chloride, cool, and add 0.45 g of so-
steam into F. Simultaneously, heat A, and maintain the tem-
dium acetate trihydrate, 6 mL of dilute acetic acid and water
perature of the solution in A between 1359 C and 1459C.
to make 150 mL. Perform the test, using 50 mL of this solu-
Adjust the distilling rate to about 10 mL per minute. Collect
tion as the test solution. Prepare the control solution with
about 170 mL of the distillate, then stop the distillation,
2.0 mL of Standard Lead Solution, 0.15 g of hydroxylam-
wash C with a small quantity of water, combine the washings
monium chloride, 0.15 g of sodium acetate trihydrate, 2 mL
with the distillate, add water to make exactly 200 mL, and
of dilute acetic acid and water to make 50 mL (not more
use this solution as the test solution. Perform the test with
than 40 ppm).
the test solution as directed in the procedure of determina-
(5) Arsenic <1.11>—To 1.0 g of Natural Aluminum Sili-
tion for fluoride under Oxygen Flask Combustion Method
cate, add 5 mL of dilute hydrochloric acid, heat gently to
<1.06>. No corrective solution is used in this procedure. The
boil while shaking well, cool rapidly, and centrifuge. Mix the
content of fluoride (F) is not more than 0.01z.
residue with 5 mL of dilute hydrochloric acid with shaking,
centrifuge, then add 10 mL of water to the residue, and
repeat the extraction in the same manner. Concentrate the
JP XVII Official Monographs / Amantadine Hydrochloride 405
Amount (mg) of fluoride (F: 19.00) in the test solution precipitate with two 10-mL portions of water, centrifuging
＝ amount (mg) of fluoride in 5 mL of each time, combine these washings with the filtrate, and add
the standard solution ammonia solution (28) dropwise until a precipitate just ap-
× AT/AS × 200/V pears. Add dropwise dilute hydrochloric acid with vigorous
shaking to redissolve the precipitate. Heat the solution with
C,
0.45 g of hydroxylammonium chloride, and after cooling,
3 hours).
add 0.45 g of sodium acetate trihydrate, 6 mL of dilute ace-
Adsorptive power To 0.10 g of Natural Aluminum Silicate tic acid and water to make 150 mL. Perform the test with 50
add 20 mL of a solution of methylene blue trihydrate (3 in mL of this solution as the test solution. Prepare the control
2000), shake for 15 minutes, allow to stand for 5 hours at solution with 3.0 mL of Standard Lead Solution, 0.15 g of
37 ± 29 C, and centrifuge. Dilute 1.0 mL of the supernatant hydroxylammonium chloride, 0.15 g of sodium acetate trihy-
liquid with water to 200 mL. Place 50 mL of the solution in a drate, 2 mL of dilute acetic acid and water to make 50 mL
Nessler tube and observe horizontally or vertically against a (not more than 30 ppm).
white background: the color of the solution is not deeper (5) Arsenic <1.11>—To 1.0 g of Synthetic Aluminum Sili-
than that of the following control solution. cate add 10 mL of dilute hydrochloric acid, heat gently to
Control solution: Dilute 1.0 mL of a solution of methy- boiling while shaking well, cool rapidly, and centrifuge. Mix
lene blue trihydrate (3 in 2000) with water to 400 mL, and the residue with 5 mL of dilute hydrochloric acid with shak-
use 50 mL of this solution. ing, centrifuge, then add 10 mL of water to the residue, and
repeat the extraction in the same manner. Concentrate the
combined extracts on a water bath to 5 mL. Use this solution
ers.
as the test solution, and perform the test (not more than
2 ppm).
Synthetic Aluminum Silicate Loss on drying <2.41> Not more than 20.0z (1 g, 1059C,
3 hours).
合成ケイ酸アルミニウム
Acid-consuming capacity <6.04> Weigh accurately about
1 g of Synthetic Aluminum Silicate, transfer to a glass-
Description Synthetic Aluminum Silicate occurs as a white stoppered flask, add 200 mL of 0.1 mol/L hydrochloric
powder. It is odorless and tasteless. acid VS, exactly measured, stopper the flask, and shake at
It is practically insoluble in water, in ethanol (95) and in 37 ± 29C for 1 hour. Filter, pipet 50 mL of the filtrate, and
diethyl ether. titrate <2.50> by stirring well the excess hydrochloric acid
Synthetic Aluminum Silicate (1 g) dissolves when heated in with 0.1 mol/L sodium hydroxide VS until the pH of the so-
20 mL of a solution of sodium hydroxide (1 in 5), leaving a lution changes to 3.5. The volume of 0.1 mol/L hydrochloric
small amount of insoluble substance . acid VS consumed is not less than 50.0 mL per g of Synthetic
Aluminum Silicate.
Identification (1) To 0.5 g of Synthetic Aluminum Sili-
cate add 3 mL of diluted sulfuric acid (1 in 3), heat until Containers and storage Containers—Well-closed contain-
white fumes evolve, cool, add 20 mL of water, and filter. ers.
Render the filtrate slightly acid with ammonia TS: the solu-
tion responds to the Qualitative Tests <1.09> for aluminum
salt. Amantadine Hydrochloride
(2) Prepare a bead by fusing ammonium sodium
hydrogenphosphate tetrahydrate on a platinum loop. Place アマンタジン塩酸塩
the bead in contact with Synthetic Aluminum Silicate, and
fuse again: an infusible material appears in the bead,
producing, upon cooling, an opaque bead with a web-like
structure.
Purity (1) Acidity or alkalinity—Shake 1.0 g of Synthetic
Aluminum Silicate with 20 mL of water, and centrifuge: the
C10H17N.HCl: 187.71
supernatant liquid so obtained is neutral.
Tricyclo[3.3.1.13,7]dec-1-ylamine monohydrochloride
(2) Chloride <1.03>—To 5.0 g of Synthetic Aluminum
[665-66-7]
Silicate add 100 mL of water, boil gently for 15 minutes
while shaking, then cool, add water to restore the original
Amantadine Hydrochloride, when dried, contains
volume, and centrifuge. To 10 mL of the supernatant liquid
not less than 99.0z of amantadine hydrochloride
add 6 mL of dilute nitric acid and water to make 50 mL, and
(C10H17N.HCl).
pare the control solution with 0.30 mL of 0.01 mol/L hydro- Description Amantadine Hydrochloride occurs as a white,
chloric acid VS (not more than 0.021z). crystalline powder. It is odorless, and has a bitter taste.
(3) Sulfate <1.14>—To 2.0 mL of the supernatant liquid It is very soluble in formic acid, freely soluble in water, in
obtained in (2) add 1 mL of dilute hydrochloric acid and methanol and in ethanol (95), and practically insoluble in
water to make 50 mL. Perform the test using this solution as diethyl ether.
the test solution. Prepare the control solution with 1.0 mL of
Identification (1) To 0.1 g of Amantadine Hydrochloride
0.005 mol/L sulfuric acid VS (not more than 0.480z).
add 1 mL of pyridine and 0.1 mL of acetic anhydride, dis-
(4) Heavy metals <1.07>—To 3.0 g of Synthetic Alumi-
solve by boiling for 1 minute, add 10 mL of dilute hydro-
num Silicate add 50 mL of water and 5 mL of hydrochloric
chloric acid, and cool in ice water. Filter the crystals sepa-
acid, boil gently for 20 minutes while shaking, then after
rated, wash with water, and dry at 1059 C for 1 hour: the
cooling, centrifuge, remove the supernatant liquid, wash the
406 Ambenonium Chloride / Official Monographs JP XVII
residue melts <2.60> between 1479C and 1519C. peak.
Amantadine Hydrochloride, previously dried, as directed in
3 hours).
photometry <2.25>, and compare the spectrum with the Ref- Residue on ignition <2.44> Not more than 0.2z (1 g).
Assay Weigh accurately about 0.2 g of Amantadine Hy-
drochloride, previously dried, dissolve in 2 mL of formic
(3) A solution of Amantadine Hydrochloride (1 in 50)
acid, add exactly 15 mL of 0.1 mol/L perchloric acid VS,
and heat on a water bath for 30 minutes. After cooling, add
pH <2.54> Dissolve 1.0 g of Amantadine Hydrochloride in acetic acid (100) to make 70 mL, and titrate <2.50> the excess
5 mL of water: the pH of this solution is between 4.0 and perchloric acid with 0.1 mol/L sodium acetate VS (potentio-
6.0. metric titration). Perform a blank determination, and make
any necessary correction.
of Amantadine Hydrochloride in 10 mL of water: the solu- Each mL of 0.1 mol/L perchloric acid VS
tion is clear and colorless. ＝ 18.77 mg of C10H17N.HCl
(2) Heavy metals <1.07>—Proceed with 2.0 g of Amanta-
dine Hydrochloride according to Method 4, and perform the
ers.
test. Prepare the control solution with 2.0 mL of Standard
Lead Solution (not more than 10 ppm).
of Amantadine Hydrochloride according to Method 3, and Ambenonium Chloride
アンベノニウム塩化物
(4) Related substances—Dissolve 0.50 g of Amantadine
Hydrochloride in 10 mL of water, add 10 mL of sodium hy-
droxide TS and 10 mL of chloroform, and shake. Filter the
chloroform layer through absorbent cotton with 3 g of anhy-
drous sodium sulfate on a funnel, and use the filtrate as the
sample solution. Pipet 1 mL of the sample solution, add
chloroform to make exactly 100 mL, and use this solution as
the standard solution. Perform the test with exactly 2 mL C28H42Cl4N4O2: 608.47
each of the sample solution and standard solution as directed 2,2?-[(1,2-Dioxoethane-1,2-diyl)diimino]bis[N-
under Gas Chromatography <2.02> according to the follow- (2-chlorobenzyl)-N, N-diethylethylaminium] dichloride
ing conditions. Determine each peak area of these solutions [115-79-7]
by the automatic integration method: each peak area other
than amantadine from the sample solution is not larger than Ambenonium Chloride contains not less than 98.5z
1/3 times the peak area of amantadine from the standard so- of ambenonium chloride (C28H42Cl4N4O2), calculated
lution, and the total area of each peak is not larger than the on the dried basis.
peak area of amantadine from the standard solution.
Description Ambenonium Chloride occurs as a white pow-
der.
Detector: A hydrogen flame-ionization detector.
It is freely soluble in water, in methanol and in acetic acid
Column: A glass column about 3 mm in inside diameter
(100), soluble in ethanol (95), and slightly soluble in acetic
and about 2 m in length, packed with siliceous earth for gas
anhydride.
chromatography (150 to 180 mm in particle diameter) coated
It is hygroscopic.
with a mixture (L) of branched hydrocarbon of petroleum
hexamethyltetracosane group for gas chromatography and
potassium hydroxide at the ratios of 2z and 1z, respec- Identification (1) Determine the absorption spectrum of a
tively. solution of Ambenonium Chloride in methanol (1 in 5000) as
Column temperature: Inject at a constant temperature of directed under Ultraviolet-visible Spectrophotometry <2.24>,
about 1259C, maintain the temperature for 5 minutes, raise and compare the spectrum with the Reference Spectrum:
at the rate of 59C per minute to 1509 C, and maintain at a both spectra exhibit similar intensities of absorption at the
constant temperature of about 1509 C for 15 minutes. same wavelengths.
Carrier gas: Nitrogen. (2) Determine the infrared absorption spectrum of
Flow rate: Adjust so that the retention time of amantadine Ambenonium Chloride, previously dried, as directed in the
is about 11 minutes. potassium chloride disk method under Infrared Spectropho-
Selection of column: Dissolve 0.15 g of naphthalene in 5 tometry <2.25>, and compare the spectrum with the Refer-
mL of the sample solution, and add chloroform to make 100 ence Spectrum: both spectra exhibit similar intensities of
mL. Proceed with 2 mL of this solution under the above absorption at the same wave numbers.
operating conditions, and calculate the resolution. Use a (3) A solution of Ambenonium Chloride (1 in 100)
column giving elution of naphthalene and amantadine in this responds to the Qualitative Tests <1.09> for chloride.
than 2.5.
of Ambenonium Chloride in 10 mL of water: the solution is
that the peak height of amantadine obtained from 2 mL of
(2) Heavy metals <1.07>—Proceed with 1.0 g of Am-
the standard solution composes about 10z of the full scale.
benonium Chloride according to Method 4, and perform the
Time span of measurement: About twice as long as the
test. Use a solution of magnesium nitrate in ethanol (95)
retention time of amantadine, beginning after the solvent
JP XVII Official Monographs / Amidotrizoic Acid 407
(1 in 5). Prepare the control solution with 2.0 mL of Stand- (2) Primary aromatic amines—Dissolve 0.20 g of
ard Lead Solution (not more than 20 ppm). Amidotrizoic Acid in 5 mL of water and 1 mL of sodium
(3) Related substances—Dissolve 0.10 g of Ambenonium hydroxide TS, add 4 mL of a solution of sodium nitrite (1 in
Chloride in 10 mL of methanol, and use this solution as the 100) and 10 mL of 1 mol/L hydrochloric acid TS, shake, and
sample solution. Pipet 1 mL of the sample solution, and add allow to stand for 2 minutes. Add 5 mL of ammonium
methanol to make exactly 20 mL. Pipet 1 mL of this solu- amidosulfate TS, shake well, allow to stand for 1 minute,
tion, add methanol to make exactly 10 mL, and use this solu- and add 0.4 mL of a solution of 1-naphthol in ethanol (95)
tion as the standard solution. Perform the test with these so- (1 in 10), 15 mL of sodium hydroxide TS and water to make
lutions as directed under Thin-layer Chromatography <2.03>. exactly 50 mL. Determine the absorbance of this solution at
Spot 5 mL each of the sample solution and standard solution 485 nm as directed under Ultraviolet-visible Spectropho-
on a plate of silica gel for thin-layer chromatography. De- tometry <2.24> using a solution, prepared in the same man-
velop the plate with a mixture of 1-butanol, formic acid and ner, as the blank: the absorbance is not more than 0.15.
water (12:6:5) to a distance of about 10 cm, and air-dry the (3) Soluble halides—Dissolve 2.5 g of Amidotrizoic Acid
plate. Allow the plate to stand in iodine vapor: the spots in 20 mL of water and 2.5 mL of ammonia TS, add 20 mL of
other than the principal spot from the sample solution are dilute nitric acid and water to make 100 mL, allow to stand
not more intense than the spot from the standard solution. for 15 minutes with occasional shaking, and filter. Discard
the first 10 mL of the filtrate, transfer the subsequent 25 mL
C,
of the filtrate to a Nessler tube, and add ethanol (95) to
4 hours).
make 50 mL. Proceed as directed under Chloride Limit Test
Residue on ignition <2.44> Not more than 0.2z (1 g). <1.03> using this solution as the test solution. Prepare the
control solution as follows: to 0.10 mL of 0.01 mol/L hydro-
Assay Weigh accurately about 0.3 g of Ambenonium Chlo-
chloric acid VS, add 6 mL of dilute nitric acid and water to
ride, and dissolve in 50 mL of a mixture of acetic anhydride
make 25 mL, then ethanol (95) to make 50 mL.
and acetic acid (100) (7:3). Titrate <2.50> with 0.1 mol/L per-
(4) Iodine—Dissolve 0.20 g of Amidotrizoic Acid in 2.0
chloric acid VS (potentiometric titration). Perform a blank
mL of sodium hydroxide TS, add 2.5 mL of 0.5 mol/L sul-
determination, and make any necessary correction.
furic acid TS, allow to stand for 10 minutes with occasional
Each mL of 0.1 mol/L perchloric acid VS shaking, add 5 mL of chloroform, shake well, and allow to
＝ 30.42 mg of C28H42Cl4N4O2 stand: the solution is colorless in the chloroform layer.
Amidotrizoic Acid according to Method 2, and perform the
Amidotrizoic Acid (6) Arsenic <1.11>—Prepare the test solution with 0.6 g
of Amidotrizoic Acid according to Method 3, and perform
アミドトリゾ酸
the test (not more than 3.3 ppm).
4 hours).
Assay Transfer about 0.5 g of Amidotrizoic Acid, accu-
rately weighed, to a saponification flask, dissolve in 40 mL
of sodium hydroxide TS, add 1 g of zinc powder, connect to
C11H9I3N2O4: 613.91 a reflux condenser, boil for 30 minutes, cool, and filter.
3,5-Bis(acetylamino)-2,4,6-triiodobenzoic acid Wash the flask and the filter paper with 50 mL of water, and
[117-96-4] combine the washings and the filtrate. Add 5 mL of acetic
acid (100) to this solution, and titrate <2.50> with 0.1 mol/L
Amidotrizoic Acid, calculated on the dried basis, silver nitrate VS until the color of the precipitate changes
contains not less than 98.0z of amidotrizoic acid from yellow to green (indicator: 1 mL of tetrabromophenol-
(C11H9I3N2O4). phthalein ethyl ester TS).
Description Amidotrizoic Acid occurs as a white crystalline Each mL of 0.1 mol/L silver nitrate VS
powder. It is odorless. ＝ 20.46 mg of C11H9I3N2O4
It is slightly soluble in ethanol (95), very slightly soluble in
water, and practically insoluble in diethyl ether.
Identification (1) Heat 0.1 g of Amidotrizoic Acid over a
flame: a purple gas is evolved.
Amidotrizoic Acid as directed in the potassium bromide disk
numbers.
of Amidotrizoic Acid in 10 mL of 0.2 mol/L sodium hydrox-
ide TS: the solution is clear and colorless.
408 Amikacin Sulfate / Official Monographs JP XVII
Amikacin Sulfate according to Method 2, and perform the
Amikacin Sulfate test. Prepare the control solution with 2.0 mL Standard
アミカシン硫酸塩 (2) Related substances—Dissolve 0.10 g of Amikacin
Sulfate in 4 mL of a water, and use this solution as the sam-
ple solution. Pipet 1 mL of the sample solution, add water to
under Thin-layer chromatography <2.03>. Spot 2 mL each of
the sample solution and standard solution on a plate of silica
gel for thin-layer chromatography. Develop the plate with a
mixture of water, ammonia water (28), methanol and tetra-
hydrofuran (1:1:1:1) to a distance of about 10 cm, and air-
dry the plate. Spray evenly ninhydrin-citric acid-acetic acid
TS on the plate, and heat at 1009C for 10 minutes: the spots
other than the principal spot from the sample solution are
not more intense than the spot from the standard solution.
um, 609C, 3 hours).
C22H43N5O13.2H2SO4: 781.76
Assay Weigh accurately an amount of Amikacin Sulfate
3-Amino-3-deoxy-a-D-glucopyranosyl-(1→6)-
and Amikacin Sulfate RS, equivalent to about 50 mg (po-
[6-amino-6-deoxy-a-D-glucopyranosyl-(1→4)]-1-N-
tency), dissolve each in water to make exactly 50 mL. Pipet
[(2S )-4-amino-2-hydroxybutanoyl]-2-deoxy-D-streptamine
200 mL each of these solutions in the test tube with glass
disulfate
stopper, add exactly 3 mL of pyridine and exactly 2 mL of a
[39831-55-5]
solution of 2,4,6-trinitrobenzenesulfonic acid (1 in 100),
stopper tightly, and heat in a water bath at 709 C for 30
Amikacin Sulfate is the sulfate of a derivative of
minutes. After cooling, add exactly 2 mL each of acetic aid
kanamycin.
(100), and use these solutions as the sample solution and
standard solution, respectively. Perform the test with exactly
20 mL each of these solutions as directed under Liquid Chro-
dried basis. The potency of Amikacin Sulfate is ex-
matography <2.01> according to the following conditions,
pressed as mass (potency) of amikacin (C22H43N5O13:
and determine the heights, HT and HS, of the peak of amika-
585.60).
cin derivative in each solution.
Description Amikacin Sulfate occurs as a white to yellow-
Amount [ mg (potency)] of amikacin (C22H43N5O13)
ish white powder.
＝ MS × HT/HS × 1000
It is very soluble in water, and practically insoluble in
ethanol (95). MS: Amount [mg (potency)] of Amikacin Sulfate RS
taken
trum of Amikacin Sulfate, previously dried, as directed in Operating conditions—
the potassium bromide disk method under Infrared Spectro- Detector: An ultraviolet absorption photometer (wave-
photometry <2.25>, and compare the spectrum with the Ref- length: 340 nm).
erence Spectrum or the spectrum of Amikacin Sulfate RS Column: A stainless steel column 4.6 mm in inside diame-
previously dried: both spectra exhibit similar intensities of ter and 25 cm in length, packed with octadecylsilanized silica
absorption at the same wave numbers. gel for liquid chromatography (5 mm in particle diameter).
(2) Dissolve 0.1 g each of Amikacin Sulfate and Amika- Column temperature: A constant temperature of about
cin Sulfate RS in 4 mL of water, and use these solutions as 359C.
the sample solution and standard solution. Perform the test Mobile phase: Dissolve 2.72 g of potassium dihydrogen-
with these solutions as directed under Thin-layer chromatog- phosphate in 800 mL of water, adjust to pH 6.5 with a solu-
raphy <2.03>. Spot 2 mL each of the sample solution and tion of potassium hydroxide (1 in 40), and add water to
standard solution on a plate of silica gel for thin-layer chro- make 1000 mL. To 280 mL of this solution add 720 mL of
matography. Develop the plate with a mixture of water, methanol, and mix.
ammonia water (28), methanol and tetrahydrofuran (1:1:1:1) Flow rate: Adjust so that the retention time of amikacin
to a distance of about 10 cm, and air-dry the plate. Spray derivative is about 9 minutes.
evenly ninhydrin-citric acid-acetic acid TS on the plate, and System suitability—
heat at 1009C for 10 minutes: the principal spot obtained System performance: Dissolve about 5 mg (potency) of
from the sample solution and the spot from the standard so- Amikacin Sulfate and about 5 mg (potency) of Kanamycin
lution show a red-purple color and the same R f value. Sulfate in 5 mL of water. Transfer 200 mL of this solution in
(3) A solution of Amikacin Sulfate (1 in 100) responds to a glass-stopperd test tube, add 3 mL of pyridine and 2 mL of
the Qualitative Tests <1.09> (1) for sulfate. a solution of 2,4,6-trinitrobenzenesulfonic acid (1 in 100),
stopper tightly, heat in a water bath at 709C for 30 minutes.
D : ＋76 – ＋849(1 g, water, 100
After cooling, add 2 mL of acetic acid (100). When the
mL, 100 mm).
procedure is run with 20 mL of this solution under the above
pH <2.54> Dissolve 1.0 g of Amikacin Sulfate in 100 mL of operating conditions, amikacin derivative and kanamycin
water: the pH of the solution is between 6.0 and 7.5. derivative are eluted in this order with the resolution between
JP XVII Official Monographs / Amikacin Sulfate for Injection 409

with 20 mL of the standard solution under the above operat- Amikacin Sulfate for Injection
ing conditions, the relative standard deviation of the ratios
of the peak height of amikacin derivative is not more than 注射用アミカシン硫酸塩
2.0z.
Containers and storage Containers—Hermetic containers. Amikacin Sulfate for Injection is a preparation for
injection, which is dissolved before use.
Amikacin Sulfate Injection than 115.0z of the labeled potency of amikacin
(C22H43N5O13: 585.60).
アミカシン硫酸塩注射液
tions, with Amikacin Sulfate.
Amikacin Sulfate Injection is an aqueous injection.
Description Amikacin Sulfate for Injection occurs as white
to yellowish white masses or powder.
than 115.0z of the labeled potency of amikacin
(C22H43N5O13: 585.60). Identification Dissolve an amount of Amikacin Sulfate for
Injection, equivalent to 25 mg (potency) of Amikacin Sul-
fate, in 1 mL of water, and use this solution as the sample
tions, with Amikacin Sulfate.
solution. Separately, dissolve 25 mg (potency) of Amikacin
Description Amikacin Sulfate Injection occurs as a color- Sulfate RS in 1 mL of water, and use this solution as the
less or pale yellow clear liquid. standard solution. Then, proceed as directed in the Identifi-
cation (2) under Amikacin Sulfate.
Identification To a volume of Amikacin Sulfate Injection,
equivalent to 0.1 g (potency) of Amikacin Sulfate, add water Osmotic pressure ratio Being specified separately when the
to make 4 mL, and use this solution as the sample solution. drug is granted approval based on the Law.
Separately, dissolve 25 mg (potency) of Amikacin Sulfate RS
pH <2.54> Dissolve an amount of Amikacin Sulfate for In-
in 1 mL of water, and use this solution as the standard solu-
jection, equivalent to 0.1 g (potency) of Amikacin Sulfate, in
tion. Then, proceed as directed in the Identifiction (2) under
10 mL of water: the pH of this solution is 6.0 to 7.5.
Amikacin Sulfate.
Osmotic pressure ratio Being specified separately when the
of Amikacin Sulfate for Injection, equivalent to 0.5 g (po-
drug is granted approval based on the Law.
tency) of Amikacin Sulfate, in 5 mL of water: the solution is
pH <2.54> 6.0 – 7.5 clear, and the absorbance at 405 nm of the solution deter-
mined as directed under Ultraviolet-visible Spectrophotome-
Bacterial endotoxins <4.01> Less than 0.50 EU/mg (po-
try <2.24> is not more than 0.15.
tency).
um, 609C, 3 hours).
tency).
ment.
brane filtration method: it meets the requirement.
Assay Take exactly a volume of Amikacin Sulfate Injec-
tion, equivalent to about 0.1 g (potency) of Amikacin Sul-
ment.
fate, and add water to make exactly 100 mL. Separately,
weigh accurately an amount of Amikacin Sulfate RS, Sterility <4.06> Perform the test according to the Mem-
equivalent to about 50 mg (potency), and add water to make brane filtration method: it meets the requirement.
exactly 50 mL. Take exactly 200 mL each of these solutions
Assay Weigh accurately the mass of the content of not less
into stoppered test tubes, then proceed as directed in the
than 10 Amikacin Sulfate for Injection. Weigh accurately a
Assay under Amikacin Sulfate.
portion of the content, equivalent to about 50 mg (potency)
Amount [mg (potency)] of amikacin (C22H43N5O13) of Amikacin Sulfate, dissolve in water to make exactly 50
＝ M S × HT / HS × 2 mL. Separately, weigh accurately an amount of Amikacin
Sulfate RS, equivalent to about 50 mg (potency), and dis-
MS: Amount [mg (potency)] of Amikacin Sulfate RS
solve in water to make exactly 50 mL. Transfer exactly 200
taken
mL each of these solutions to separate glass stoppered tubes,
Containers and storage Containers—Hermetic containers. and proceed as directed in the Assay under Amikacin Sul-
fate.
Amount [mg (potency)] of amikacin (C22H43N5O13)
＝ MS × HT/HS
MS: Amount [mg (potency)] of Amikacin Sulfate RS
taken
410 Aminophylline Hydrate / Official Monographs JP XVII
Containers and storage Containers—Hermetic containers. of Aminophylline Hydrate in 10 mL of hot water: the solu-
tion is clear and colorless to pale yellow.
Aminophylline Hydrate Aminophylline Hydrate according to Method 2, and per-
アミノフィリン水和物 Standard Lead Solution (not more than 20 ppm).
Water <2.48> Not more than 7.9z (0.3 g, direct titration).
Assay (1) Theophylline—Weigh accurately about 0.25 g
of Aminophylline Hydrate, and dissolve in 50 mL of water
and 8 mL of ammonia TS by gentle warming on a water
bath. Add exactly 20 mL of 0.1 mol/L silver nitrate VS,
C14H16N8O4.C2H8N2.xH2O
warm on a water bath for 15 minutes, allow to stand be-
1,3-Dimethyl-1H-purine-2,6(3H,7H )-dione
tween 59 C and 109C for 20 minutes, collect the precipitate
hemi(ethane-1,2-diamine) hydrate
by suction, and wash with three 10-mL portions of water.
[76970-41-7, monohydrate]
Combine the filtrate and washings, and add dilute nitric acid
to make neutral. Add 3 mL of dilute nitric acid, and titrate
Aminophylline Hydrate contains not less than <2.50> the excess silver nitrate with 0.1 mol/L ammonium
84.0z and not more than 86.0z of theophylline
thiocyanate VS (indicator: 2 mL of ammonium iron (III)
(C7H8N4O2: 180.16), and not less than 14.0z and not
sulfate TS). Perform a blank determination.
more than 15.0z of ethylenediamine (C2H8N2: 60.10),
calculated on the anhydrous basis. Each mL of 0.1 mol/L silver nitrate VS
＝ 18.02 mg of C7H8N4O2
Description Aminophylline Hydrate occurs as white to pale
yellow, granules or powder. It is odorless or slightly ammo- (2) Ethylenediamine—Weigh accurately about 0.5 g of
nia-like odor, and has a bitter taste. Aminophylline Hydrate, dissolve in 30 mL of water, and
It is soluble in water, slightly soluble in methanol, and titrate <2.50> with 0.1 mol/L hydrochloric acid VS (indica-
practically insoluble in ethanol (95) and in diethyl ether. tor: 3 drops of bromophenol blue TS).
To 1 g of Aminophylline Hydrate add 5 mL of water, and
Each mL of 0.1 mol/L hydrochloric acid VS
shake: it dissolves almost completely. Separation of crystals
＝ 3.005 mg of C2H8N2
begins in 2 to 3 minutes, and these crystals dissolve on the
addition of a small amount of ethylenediamine. Containers and storage Containers—Tight containers.
It is gradually affected by light, and gradually loses ethyl- Storage—Light-resistant.
enediamine in air.
Identification (1) Dissolve 0.75 g of Aminophylline Hy-
drate in 30 mL of water, and use this solution as the sample Aminophylline Injection
solution. To 20 mL of the sample solution add 1 mL of
アミノフィリン注射液
dilute hydrochloric acid: a precipitate is gradually formed.
Filter the precipitate, recrystallize from water, and dry at
1059C for 1 hour: the crystals so obtained melt <2.60> Aminophylline Injection is an aqueous injection.
between 2719C and 2759C. It contains not less than 75.0z and not more
(2) Dissolve 0.1 g of the crystals obtained in (1) in 50 mL than 86.0z of the labeled amount of theophylline
of water, and to 2 mL of this solution add tannic acid TS (C7H8N4O2: 180.16), and not less than 13.0z and not
dropwise: a white precipitate is produced, and this precipi- more than 20.0z of ethylenediamine (C2H8N2: 60.10).
tate dissolves upon dropwise addition of tannic acid TS. The concentration of Aminophylline Injection is ex-
(3) To 0.01 g of the crystals obtained in (1) add 10 drops pressed as the quantity of aminophylline dihydrate
of hydrogen peroxide TS and 1 drop of hydrochloric acid, (C16H24N10O4.2H2O: 456.46).
and evaporate on a water bath to dryness: the residue shows
a yellow-red color. Invert the dish containing the residue
tions, with Aminophylline Hydrate. It may be prepared with
over a vessel containing 2 to 3 drops of ammonia TS: the
Theophylline and its equivalent Ethylenediamine, instead of
color of the residue changes to red-purple, which is des-
Aminophylline Hydrate.
troyed on the addition of 2 to 3 drops of sodium hydroxide
It may contain not more than 60 mg of Ethylenediamine
TS.
as a stabilizer for each g of Aminophylline Hydrate.
(4) Dissolve 0.01 g of the crystals obtained in (1) in 5 mL
of water, add 3 mL of ammonia-ammonium chloride buffer Description Aminophylline Injection is a clear and color-
solution (pH 8.0) and 1 mL of copper (II) sulfate-pyridine less liquid. It has a slightly bitter taste.
TS, and mix. Add 5 mL of chloroform to the mixture, and It gradually changes in color by light.
shake: the chloroform layer develops a green color. pH: 8.0 – 10.0
(5) To 5 mL of the sample solution obtained in (1) add 2
Identification To a volume of Aminophylline Injection,
drops of copper (II) sulfate TS: a purple color develops. Add
equivalent to 0.75 g of Aminophylline Hydrate, add water to
1 mL of copper (II) sulfate TS: the color changes to blue,
make 30 mL. Proceed with this solution as directed in the
and green precipitates are formed on standing.
Identification under Aminophylline Hydrate.
pH <2.54> Dissolve 1.0 g of Aminophylline Hydrate in 25
Bacterial endotoxins <4.01> Less than 0.6 EU/mg.
mL of water: the pH of the solution is between 8.0 and 9.5.
JP XVII Official Monographs / Amiodarone Hydrochloride 411

to Method 1: it meets the requirement. Amiodarone Hydrochloride
アミオダロン塩酸塩
ment.
Assay (1) Theophylline—Pipet a volume of Aminophyl-
line Injection, equivalent to about 39.4 mg of theophylline
(C7H8N4O2) (about 50 mg of Aminophylline Hydrate), add
water to make exactly 50 mL, and use this solution as the C25H29I2NO3.HCl: 681.77
sample solution. Separately, weigh accurately about 40 mg (2-Butylbenzofuran-3-yl){4-[2-(diethylamino)ethoxy]-3,5-
of theophylline for assay, previously dried at 1059 C for 4 diiodophenyl}methanone monohydrochloride
hours, dissolve in water to make exactly 50 mL, and use this [19774-82-4]
solution as the standard solution. Perform the test with ex-
actly 5 mL each of the sample solution and standard solution Amiodarone Hydrochloride, when dried, contains
as directed under Liquid Chromatography <2.01> according not less than 98.5z and not more than 101.0z of
to the following conditions, and determine the peak areas, amiodarone hydrochloride (C25H29I2NO3.HCl).
AT and AS, of theophylline in each solution.
Description Amiodarone Hydrochloride occurs as a white
Amount (mg) of theophylline (C7H8N4O2) to pale yellowish white crystalline powder.
＝ M S × AT / AS It is very soluble in water at 809 C, freely soluble in
dichloromethane, soluble in methanol, sparingly soluble in
MS: Amount (mg) of theophylline for assay taken
ethanol (95), and very slightly soluble in water.
Operating conditions— Melting point: about 1619C (with decomposition).
length: 270 nm).
solution of Amiodarone Hydrochloride in ethanol (95) (1 in
Column: A stainless steel column 6 mm in inside diameter
and 15 cm in length, packed with octadecylsilanized silica gel
ence Spectrum: both spectra exhibit similar intensities of ab-
sorption at the same wavelengths.
409 C.
Amiodarone Hydrochloride as directed in the potassium
100) and methanol (4:1).
Flow rate: Adjust so that the retention time of theophyl-
line is about 5 minutes.
the same wave numbers.
(3) To 0.1 g of Amiodarone Hydrochloride add 10 mL
of water, dissolve by warming at 809C, and cool: the solu-
tions, the number of theoretical plates and the symmetry fac-
tion responds to the Qualitative Tests <1.09> (2) for chloride.
tor of the peak of theophylline are not less than 3000 and not
more than 1.5, respectively. pH <2.54> To 1.0 g of Amiodarone Hydrochloride add 20
System repeatability: When the test is repeated 6 times mL of freshly boiled and cooled water, dissolve by warming
with 5 mL of the standard solution under the above operating at 809C, and cool: the pH of this solution is between 3.2 and
conditions, the relative standard deviation of the peak area 3.8.
of theophylline is not more than 1.0z.
(2) Ethylenediamine—To an accurately measured
of Amiodarone Hydrochloride in 10 mL of methanol: the
volume of Aminophylline Injection, equivalent to about 30
solution is clear, and is not more colored than the following
mg of ethylenediamine (C2H8N2) (about 0.2 g of Aminophyl-
control solutions (1) and (2).
line Hydrate), add water to make 30 mL, and titrate <2.50>
Control solution (1): To a mixture of 1.0 mL of Cobalt
with 0.1 mol/L hydrochloric acid VS (indicator: 2 to 3 drops
(II) Chloride CS, 2.4 mL of Iron (III) Chloride CS and 0.4
of bromophenol blue TS).
mL of Copper (II) Sulfate CS, add diluted hydrochloric acid
Each mL of 0.1 mol/L hydrochloric acid VS (1 in 40) to make 10.0 mL. To 2.5 mL of this solution add
＝ 3.005 mg of C2H8N2 diluted hydrochloric acid (1 in 40) to make 20 mL.
Control solution (2): To 3.0 mL of a mixture of 0.2 mL of
Cobalt (II) Chloride CS, 9.6 mL of Iron (III) Chloride CS
Plastic containers for aqueous injections may be used.
and 0.2 mL of Copper (II) Sulfate CS, add diluted hydro-
chloric acid (1 in 40) to make 100 mL.
(2) Iodine—To 1.50 g of Amiodarone Hydrochloride
add 40 mL of water, dissolve by warming at 809C, cool, add
water to make exactly 50 mL, and use this solution as the
sample stock solution. Pipet 15 mL of this solution, add
exactly 1 mL of 0.1 mol/L hydrochloric acid TS and exactly
1 mL of a solution of potassium iodate (107 in 10,000), add
sample solution. Separately, pipet 15 mL of the sample stock
412 Amiodarone Hydrochloride Tablets / Official Monographs JP XVII
solution, add exactly 1 mL of 0.1 mol/L hydrochloric acid Flow rate: Adjust so that the retention time of amioda-
TS, exactly 1 mL of a solution of potassium iodide (441 in rone is about 24 minutes.
5,000,000) and exactly 1 mL of a solution of potassium Time span of measurement: About 2 times as long as the
iodate (107 in 10,000), add water to make exactly 20 mL, retention time of amiodarone.
and use this solution as the standard solution. Separately, System suitability—
pipet 15 mL of the sample stock solution, add exactly 1 mL Test for required detectability: Pipet 5 mL of the standard
of 0.1 mol/L hydrochloric acid TS, add water to make ex- solution, and add a mixture of water and acetonitrile for liq-
actly 20 mL, and use this solution as the control solution. uid chromatography (1:1) to make exactly 25 mL. Confirm
Allow the sample solution, standard solution and control so- that the peak area of amiodarone obtained from 10 mL of
lution to stand in a dark place for 4 hours. Perform the test this solution is equivalent to 14 to 26z of that obtained
with the sample solution and standard solution as directed from 10 mL of the standard solution.
under Ultraviolet-visible Spectrophotometry <2.24>, using System performance: When the procedure is run with 10
the control solution as the blank: the absorbance of the sam- mL of the standard solution under the above operating con-
ple solution at 420 nm is not larger than 1/2 times the absor- ditions, the number of theoretical plates and the symmetry
bance of the standard solution. factor of the peak of amiodarone are not less than 5000 and
(3) Heavy metals <1.07>—Proceed with 1.0 g of Amioda- not more than 1.5, respectively.
rone Hydrochloride according to Method 4, and perform the System repeatability: When the test is repeated 6 times
test. Prepare the control solution with 2.0 mL of Standard with 10 mL of the standard solution under the above operat-
Lead Solution (not more than 20 ppm). ing conditions, the relative standard deviation of the peak
(4) Related substance 1—Dissolve 0.5 g of Amiodarone area of amiodarone is not more than 1.0z.
Hydrochloride in 5 mL of dichloromethane, and use this
Loss on drying <2.41> Not more than 0.5z (1 g, reduced
solution as the sample solution. Separately, dissolve 10 mg
of 2-chloroethyl diethylamine hydrochloride in 50 mL of
dichloromethane, and use this solution as the standard solu- Residue on ignition <2.44> Not more than 0.1z (1 g).
tion. Perform the test with these solutions as directed under
Assay Weigh accurately about 0.6 g of Amiodarone Hy-
drochloride, previously dried, dissolve in 40 mL of a mixture
sample solution and standard solution on a plate of silica gel
of acetic anhydride and acetic acid (100) (3:1), and titrate
with fluorescent indicator for thin-layer chromatography.
Develop the plate with a mixture of dichloromethane, meth-
titration). Perform a blank determination in the same man-
anol and formic acid (17:2:1) to a distance of about 15 cm,
ner, and make any necessary correction.
and air-dry the plate. Spray evenly bismuth subnitrate TS
and then hydrogen peroxide TS: the spot obtained from the Each mL of 0.1 mol/L perchloric acid VS
sample solution corresponding to the spot from the standard ＝ 68.18 mg of C25H29I2NO3.HCl
solution is not more intense than the spot from the standard
solution.
(5) Related substance 2—Dissolve 0.125 g of Amioda-
rone Hydrochloride in 25 mL of a mixture of water and
acetonitrile for liquid chromatography (1:1), and use this so-
lution as the sample solution. Pipet 2 mL of the sample solu- Amiodarone Hydrochloride Tablets
tion, and add a mixture of water and acetonitrile for liquid
アミオダロン塩酸塩錠
chromatography (1:1) to make exactly 50 mL. Pipet 1 mL of
this solution, add a mixture of water and acetonitrile for liq-
uid chromatography (1:1) to make exactly 20 mL, and use Amiodarone Hydrochloride Tablets contain not
this solution as the standard solution. Perform the test with less than 93.0z and not more than 107.0z of
exactly 10 mL each of the sample solution and standard solu- the labeled amount of amiodarone hydrochloride
tion as directed under Liquid Chromatography <2.01> ac- (C25H29I2NO3.HCl: 681.77).
cording to the following conditions. Determine each peak
area by the automatic integration method: the area of the
with Amiodarone Hydrochloride.
peak other than amiodarone obtained from the sample solu-
tion is not larger than the peak area of amiodarone obtained Identification To 1 mL of the sample stock solution ob-
from the standard solution, and the total area of the peaks tained in the Assay add the mobile phase to make 50 mL.
other than amiodarone from the sample solution is not Determine the absorption spectrum of this solution as di-
larger than 2.5 times the peak area of amiodarone from the rected under Ultraviolet-visible Spectrophotometry <2.24>: it
standard solution. exhibits a maximum between 239 nm and 243 nm.
length: 240 nm).
To 1 tablet of Amiodarone Hydrochloride Tablets add
160 mL of the mobile phase, treat with ultrasonic waves for
10 minutes, add the mobile phase to make exactly 200 mL,
and centrifuge. Pipet V mL of the supernatant liquid,
309 C.
equivalent to about 1 mg of amiodarone hydrochloride
Mobile phase: To 800 mL of water add 3.0 mL of acetic
(C25H29I2NO3.HCl), add the mobile phase to make exactly 50
acid (100), adjust the pH to 4.95 with ammonia solution
(28), and add water to make 1000 mL. To 300 mL of this so-
weigh accurately about 25 mg of amiodarone hydrochloride
lution add 400 mL of acetonitrile for liquid chromatography
for assay, previously dried at 509 C for 4 hours under
and 300 mL of methanol for liquid chromatography.
JP XVII Official Monographs / Amiodarone Hydrochloride Tablets 413
reduced pressure not exceeding 0.3 kPa, and dissolve in the minutes, and add the mobile phase to make exactly 100 mL.
mobile phase to make exactly 50 mL. Pipet 2 mL of this so- Centrifuge this solution, and use the supernatant liquid as
lution, add the mobile phase to make exactly 50 mL, and use the sample stock solution. Pipet 2 mL of the stock solution,
this solution as the standard solution. Perform the test with add exactly 2 mL of the internal standard solution, add the
exactly 10 mL each of the sample solution and standard solu- mobile phase to make 50 mL, and use this solution as the
tion as directed under Liquid Chromatography <2.01> ac- sample solution. Separately, weigh accurately 25 mg of
cording to the following conditions, and determine the peak amiodarone hydrochloride for assay, previously dried at
areas, AT and AS, of amiodarone in each solution. 509C for 4 hours under reduced pressure not exceeding
0.3 kPa, and dissolve in the mobile phase to make exactly 50
Amount (mg) of amiodarone hydrochloride
mL. Pipet 2 mL of this solution, add exactly 2 mL of the
(C25H29I2NO3.HCl)
internal standard solution, add the mobile phase to make 50
＝ MS × AT/AS × 8/V
mL, and use this solution as the standard solution. Perform
MS: Amount (mg) of amiodarone for assay taken the test with 10 mL each of the sample solution and standard
according to the following conditions, and calculate the
ratios, QT and QS, of the peak area of amiodarone to that of
Assay.
the internal standard.
System performance: When the procedure is run with 10 Amount (mg) of amiodarone hydrochloride
mL of the standard solution under the above operating con- (C25H29I2NO3.HCl)
ditions, the number of theoretical plates and the symmetry ＝ M S × QT / QS × 2
factor of the peak of amiodarone are not less than 5000 and
MS: Amount (mg) of amiodarone hydrochloride for assay
not more than 1.5, respectively.
taken
with 10 mL of the standard solution under the above operat- Internal standard solution—A solution of chlorhexidine hy-
ing conditions, the relative standard deviation of the peak drochloride in the mobile phase (1 in 2500).
area of amiodarone is not more than 1.0z. Operating conditions—
length: 242 nm).
mL of acetic acid-sodium acetate buffer solution (pH 4.0) as
the dissolution medium, the dissolution rate in 30 minutes of
Amiodarone Hydrochloride Tablets is not less than 80z.
Start the test with 1 tablet of Amiodarone Hydrochloride
509C.
Tablets, withdraw not less than 20 mL of the medium at the
Mobile phase: A mixture of acetonitrile for liquid chroma-
specified minute after starting the test, and filter through a
tography, a solution of sodium laurylsulfate (1 in 50) and
membrane filter with a pore size not exceeding 0.45 mm. Dis-
phosphoric acid (750:250:1).
card the first 10 mL of the filtrate, pipet V mL of the subse-
Flow rate: Adjust so that the retention time of amioda-
quent filtrate, add exactly V mL of methanol, then add a
rone is about 7 minutes.
mixture of the dissolution medium and methanol (1:1) to
make exactly V? mL so that each mL contains about 11 mg of
amiodarone hydrochloride (C25H29I2NO3.HCl), and use this
ditions, the internal standard and amiodarone are eluted in
about 28 mg of amiodarone hydrochloride for assay, previ-
this order with the resolution between these peaks being not
ously dried at 509 C for 4 hours under reduced pressure not
less than 5.
exceeding 0.3 kPa, and dissolve in methanol to make exactly
50 mL. Pipet 2 mL of this solution, add exactly 2 mL of the
dissolution medium, then add a mixture of the dissolution
medium and methanol (1:1) to make exactly 100 mL, and
the peak area of amiodarone to that of the internal standard
solution at 241 nm as directed under Ultraviolet-visible Spec- Containers and storage Containers—Tight containers.
trophotometry <2.24>, using a mixture of the dissolution Storage—Light-resistant.
medium and methanol (1:1) as the blank.
of amiodarone hydrochloride (C25H29I2NO3.HCl)
＝ MS × AT/AS × V?/V × 1/C × 36
MS: Amount (mg) of amiodarone hydrochloride for assay
taken
C: Labeled amount (mg) of amiodarone hydrochloride
(C25H29I2NO3.HCl) in 1 tablet
Assay Weigh accurately the mass of not less than 20 Amio-
darone Hydrochloride Tablets, and powder. Weigh accu-
rately a portion of the powder, equivalent to about 50 mg of
amiodarone hydrochloride (C25H29I2NO3.HCl), add 80 mL
of the mobile phase, treat with ultrasonic waves for 10
414 Amitriptyline Hydrochloride / Official Monographs JP XVII
Amitriptyline Hydrochloride
アミトリプチリン塩酸塩 Amitriptyline Hydrochloride
Tablets
アミトリプチリン塩酸塩錠
Amitriptyline Hydrochloride Tablets contain not

less than 90.0z and not more than 110.0z of
C20H23N.HCl: 313.86 the labeled amount of amitriptyline hydrochloride
3-(10,11-Dihydro-5H-dibenzo[a,d ]cyclohepten-5- (C20H23N.HCl: 313.86).
ylidene)-N, N-dimethylpropylamine monohydrochloride
[549-18-8]
with Amitriptyline Hydrochloride.
Amitriptyline Hydrochloride, when dried, contains Identification (1) Weigh a quantity of powdered Ami-
not less than 99.0z of amitriptyline hydrochloride triptyline Hydrochloride Tablets, equivalent to 0.1 g of Ami-
(C20H23N.HCl). triptyline Hydrochloride. Add 10 mL of chloroform, shake
thoroughly, and filter. Evaporate the filtrate on a water bath
Description Amitriptyline Hydrochloride occurs as color-
to about 2 mL, add diethyl ether until turbidity is produced,
less crystals or a white to pale yellow crystalline powder. It
and allow to stand. Filter the crystals formed through a glass
has a bitter taste and a numbing effect.
filter (G4), and proceed as directed in the Identification (1)
It is freely soluble in water, in ethanol (95) and in acetic
and (2) under Amitriptyline Hydrochloride.
acid (100), soluble in acetic anhydride, and practically in-
soluble in diethyl ether.
the crystals obtained in (1) (1 in 100,000) as directed under
The pH of a solution of 1.0 g of Amitriptyline Hydrochlo-
ride in 20 mL of water is between 4.0 and 5.0.
maximum between 238 nm and 240 nm, and a minimum be-
Identification (1) Dissolve 5 mg of Amitriptyline Hydro- tween 228 nm and 230 nm.
chloride in 3 mL of sulfuric acid: a red color develops. Add 5
drops of potassium dichromate TS to this solution: it turns
dark brown.
(2) Acidify 1 mL of a solution of Amitriptyline Hydro-
To 1 tablet of Amitriptyline Hydrochloride Tablets add 50
chloride (1 in 500) with 0.5 mL of dilute nitric acid, and add
mL of diluted methanol (1 in 2), shake to disintegrate the
1 drop of silver nitrate TS: a white, opalescent precipitate is
tablet, then add diluted methanol (1 in 2) to make exactly
produced.
100 mL, and filter. Discard the first 20 mL of the filtrate,
pipet V mL of the subsequent filtrate, add methanol to make
Amitriptyline Hydrochloride (1 in 100,000) as directed under
exactly V? mL so that each mL contains about 10 mg of
Ultraviolet-visible Spectrophotometry <2.24>, and compare
amitriptyline hydrochloride (C20H23N.HCl), and use this so-
the spectrum with the Reference Spectrum or the spectrum
lution as the sample solution. Then, proceed as directed in
of a solution of Amitriptyline Hydrochloride RS prepared in
the Assay.
the same manner as the sample solution: both spectra exhibit
similar intensities of absorption at the same wavelengths. Amount (mg) of amitriptyline hydrochloride (C20H23N.HCl)
＝ MS × AT/AS × V?/V × 1/20
MS: Amount (mg) of Amitriptyline Hydrochloride RS
taken
of Amitriptyline Hydrochloride in 20 mL of water: the solu-
tion is clear and colorless. Dissolution <6.10> When the test is performed at 50 revolu-
(2) Heavy metals <1.07>—Proceed with 2.0 g of Amitrip- tions per minute according to the Paddle method, using
tyline Hydrochloride according to Method 2, and perform 900 mL of 2nd fluid for dissolution test as the dissolution
the test. Prepare the control solution with 2.0 mL of Stand- medium, the dissolution rate in 60 minutes of Amitriptyline
ard Lead Solution (not more than 10 ppm). Hydrochloride Tablets is not less than 70z.
Start the test with 1 tablet of Amitriptyline Hydrochloride
2 hours).
Residue on ignition <2.44> Not more than 0.1z (1 g). membrane filter with a pore size not exceeding 0.8 mm.
Discard the first 10 mL of the filtrate, pipet the subsequent
Assay Weigh accurately about 0.5 g of Amitriptyline Hy-
V mL of the filtrate, add the dissolution medium to make
exactly V? mL so that each mL contains about 11 mg of
amitriptyline hydrochloride (C20H23N.HCl), and use this so-
titration). Perform a blank determination, and make any
about 55 mg of Amitriptyline Hydrochloride RS, previously
necessary correction.
dried at 1059C for 2 hours, and dissolve in the dissolution
Each mL of 0.1 mol/L perchloric acid VS medium to make exactly 250 mL. Pipet 5 mL of this solu-
＝ 31.39 mg of C20H23N.HCl tion, add the dissolution medium to make exactly 100 mL,
and use this solution as the standard solution. Determine the
absorbances, AT and AS, of the sample solution and stand-
JP XVII Official Monographs / Amlexanox 415
ard solution at 239 nm as directed under Ultraviolet-visible (2) Determine the infrared absorption spectrum of Am-
Spectrophotometry <2.24>. lexanox as directed in the potassium bromide disk method
of amitriptyline hydrochloride (C20H23N.HCl)
Amlexanox RS: both spectra exhibit similar intensities of
＝ MS × AT/AS × V?/V × 1/C × 18
MS: Amount (mg) of Amitriptyline Hydrochloride RS
Purity (1) Chloride <1.03>—Dissolve 1.0 g of Amlexanox
taken
in 20 mL of water and 10 mL of sodium hydroxide TS, add
C: Labeled amount (mg) of amitriptyline hydrochloride
15 mL of dilute nitric acid and water to make 50 mL, centri-
(C20H23N.HCl) in 1 tablet
fuge, and then filter the supernatant liquid. To 25 mL of this
Assay Weigh accurately and powder not less than 20 filtrate add water to make 50 mL. Perform the test using this
Amitriptyline Hydrochloride Tablets. Weigh accurately a solution as the test solution. The control solution consists of
portion of the powder, equivalent to about 20 mg of ami- 5 mL of sodium hydroxide TS, 7.5 mL of dilute nitric acid,
triptyline hydrochloride (C20H23N.HCl), and add 75 mL of 0.30 mL of 0.01 mol/L hydrochloric acid VS, and water ad-
diluted methanol (1 in 2). After shaking for 30 minutes, add ded to make 50 mL (not more than 0.021z).
diluted methanol (1 in 2) to make exactly 100 mL, and filter. (2) Heavy metals <1.07>—Proceed with 1.0 g of Amlexa-
Discard the first 20-mL portion of the filtrate, measure nox according to Method 2, and perform the test. Prepare
exactly the subsequent 5-mL portion, add methanol to make the control solution with 2.0 mL of Standard Lead Solution
exactly 100 mL, and use this solution as the sample solution. (not more than 20 ppm).
Separately, weigh accurately about 20 mg of Amitriptyline (3) Related substances—(i) Dissolve 30 mg of Amlexa-
Hydrochloride RS, previously dried at 1059C for 2 hours, nox in 50 mL of the mobile phase, and use this solution as
and dissolve in diluted methanol (1 in 2) to make exactly 100 the sample solution. Pipet 1 mL of the sample solution, and
mL. Measure exactly 5 mL of this solution, add methanol to add the mobile phase to make exactly 50 mL. Pipet 1 mL of
make exactly 100 mL, and use this solution as the standard this solution, add the mobile phase to make exactly 20 mL,
solution. Determine the absorbances, AT and AS, of the and use this solution as the standard solution. Perform the
sample solution and standard solution at 239 nm as directed test with exactly 10 mL each of the sample solution and
under Ultraviolet-visible Spectrophotometry <2.24>, respec- standard solution as directed under Liquid Chromatography
tively. <2.01> according to the following conditions. Determine each
peak area of these solutions by the automatic integration
Amount (mg) of amitriptyline hydrochloride (C20H23N.HCl)
method: the area of the peak other than amlexanox obtained
＝ MS × AT/AS
from the sample solution is not larger than 2 times the peak
MS: Amount (mg) of Amitriptyline Hydrochloride RS area of amlexanox obtained from the standard solution.
taken Operating conditions—
The detector, column, column temperature, mobile phase,
and flow rate: Proceed as directed in the operating condi-
tions in the Assay.
Time span of measurement: Until completion of the
Amlexanox elution of amlexanox, beginning after the solvent peak.
アンレキサノクス
Test for required detectability: Pipet 10 mL of the stand-
ard solution, and add the mobile phase to make exactly 100
mL. Confirm that the peak area of amlexanox obtained
from 10 mL of this solution is equivalent to 7 to 13z of that
C16H14N2O4: 298.29 obtained from 10 mL of the standard solution.
2-Amino-7-(1-methylethyl)-5-oxo- System repeatability: When the test is repeated 6 times
5H-[1]benzopyrano[2,3-b]pyridine-3-carboxylic acid with 10 mL of the standard solution under the above operat-
[68302-57-8] ing conditions, the relative standard deviation of the peak
area of amlexanox is not more than 2.0z.
Amlexanox, when dried, contains not less than (ii) Dissolve 30 mg of Amlexanox in 50 mL of the mobile
98.0z and not more than 102.0z of amlexanox phase, and use this solution as the sample solution. Pipet 1
(C16H14N2O4). mL of the sample solution, and add the mobile phase to
Description Amlexanox occurs as white to yellowish white,
mobile phase to make exactly 20 mL, and use this solution as
crystals or crystalline powder.
the standard solution. Perform the test with exactly 10 mL
It is very slightly soluble in ethanol (99.5), and practically
insoluble in water.
It dissolves in diluted sodium hydroxide TS (1 in 3).
lowing conditions, and determine each peak area of these so-
Identification (1) Determine the absorption spectrum of a lutions by the automatic integration method: the area of the
solution of Amlexanox in ethanol (99.5) (1 in 250,000) as peak other than amlexanox obtained from the sample solu-
directed under Ultraviolet-visible Spectrophotometry <2.24>, tion is not larger than 2 times the peak area of amlexanox
and compare the spectrum with the Reference Spectrum or obtained from the standard solution.
the spectrum of a solution of Amlexanox RS prepared in the Operating conditions—
same manner as the sample solution: both spectra exhibit Detector, column, and column temperature: Proceed as
similar intensities of absorption at the same wavelengths. directed in the operating conditions in the Assay.
416 Amlexanox Tablets / Official Monographs JP XVII
Mobile phase: Dissolve 7.2 g of disodium hydrogen phos- Operating conditions—
phate dodecahydrate in water to make 1000 mL. Adjust the Detector: An ultraviolet absorption photometer (wave-
pH of this solution to 8.0 by adding a solution prepared by length: 254 nm).
dissolving 3.1 g of sodium dihydrogen phosphate dihydrate Column: A stainless steel column 4.0 mm in inside diame-
in 1000 mL of water. To 400 mL of this solution add 600 mL ter and 15 cm in length, packed with octadecylsilanized silica
of acetonitrile. gel for liquid chromatography (5 mm in particle diameter).
Flow rate: To 15 mL of a solution of benzophenone in the Column temperature: A constant temperature of about
mobile phase (3 in 1,000,000) add the mobile phase to make 259C.
20 mL. Adjust so that the retention time of benzophenone is Mobile phase: Dissolve 17.9 g of disodium hydrogen phos-
about 6.5 minutes when perform the test with 10 mL of this phate dodecahydrate in water to make 1000 mL. Adjust the
solution under the conditions described above. pH of this solution to 8.0 by adding a solution prepared by
Time span of measurement: About 3 times as long as the dissolving 7.8 g of sodium dihydrogen phosphate dihydrate
retention time of benzophenone, beginning after the peak of in 1000 mL of water. To 760 mL of this solution add 240 mL
amlexanox. of acetonitrile.
System suitability— Flow rate: Adjust so that the retention time of amlexanox
Test for required detectability: Pipet 5 mL of the standard is about 10 minutes.
solution, and add the mobile phase to make exactly 50 mL. System suitability—
Confirm that the peak area of amlexanox obtained from System performance: When the procedure is run with 10
10 mL of this solution is equivalent to 7 to 13z of that ob- mL of the standard solution according to the above condi-
tained from 10 mL of the standard solution. tions, amlexanox and the internal standard are eluted in this
System performance: Pipet 1 mL of the sample solution, order with the resolution between these peaks being not less
and add the mobile phase to make 100 mL. To 5 mL of this than 2.0.
solution add 15 mL of the solution of benzophenone in the System repeatability: When the test is repeated 6 times
mobile phase (3 in 1,000,000). When perform the test with with 10 mL of the standard solution under the above condi-
10 mL of this solution according to the above conditions, tions, the relative standard deviation of the ratio of the peak
amlexanox and benzophenone are eluted in this order with area of amlexanox to that of the internal standard is not
the resolution between these peaks being not less than 10. more than 1.0z.
ers.
area of amlexanox is not more than 2.0z.
(iii) The total amount of related substances, when calcu-
lated according to the following formula, is not more than Amlexanox Tablets
0.5z.
アンレキサノクス錠
Total amount (z) of related substances
＝ {(AT1/AS1) ＋ (AT2/AS2)} × 1/10
Amlexanox Tablets contain not less than 93.0z
AT1: Total area of the peaks other than amlexanox from and not more than 107.0z of the labeled amount of
the sample solution obtained in (i) amlexanox (C16H14N2O4: 298.29).
AT2: Total area of the peaks other than amlexanox from
the sample solution obtained in (ii)
with Amlexanox.
AS1: Peak area of amlexanox from the standard solution
obtained in (i) Identification (1) Take an amount of powdered Amlexa-
AS2: Peak area of amlexanox from the standard solution nox Tablets, equivalent to 10 mg of Amlexanox, add 100 mL
obtained in (ii) of ethanol (99.5), shake vigorously, and filter. Pipet 1 mL of
the filtrate, add 25 mL of ethanol (99.5), and use this solu-
tion as the sample solution. Determine the absorption spec-
2 hours).
trum of the sample solution as directed under Ultraviolet-
Residue on ignition <2.44> Not more than 0.1z (1 g). visible Spectrophotometry <2.24>: it exhibits absorption
maxima between 240 nm and 244 nm, between 285 nm and
Assay Weigh accurately about 30 mg each of Amlexanox
and Amlexanox RS, both dried, and dissolve them separately
(2) Observe the sample solution obtained in (1) under
in the mobile phase to make exactly 50 mL. Pipet 5 mL each
ultraviolet light (main wavelength: 365 nm): the solution
of these solutions, and add exactly 15 mL of the internal
shows a bluish-white fluorescence.
standard solution, and use these solutions as the sample so-
lution and the standard solution, respectively. Perform the Uniformity of dosage units <6.02> Perform the Mass varia-
test with 10 mL each of the sample solution and standard so- tion test, or the Content uniformity test according to the fol-
lution as directed under Liquid Chromatography <2.01> ac- lowing method: it meets the requirement.
cording to the following conditions, and calculate the ratios, Take 1 tablet of Amlexanox Tablets, add exactly 0.6 mL
QT and QS, of the peak area of amlexanox to that of the in- of the internal standard solution per 1 mg of amlexanox
ternal standard, respectively. (C16H14N2O4), add the mobile phase to make exactly V mL
so there is about 167 mg of amlexanox (C16H14N2O4) per mL,
Amount (mg) of amlexanox (C16H14N2O4) ＝ MS × QT/QS
disintegrate the tablet, and then shake vigorously for 5
MS: Amount (mg) of Amlexanox RS taken minutes. Centrifuge this solution, and use the supernatant
liquid as the sample solution. Separately, weigh accurately
Internal standard solution—A solution of 3-nitroaniline in
about 30 mg of Amlexanox RS, previously dried at 1059C
the mobile phase (1 in 4000).
for 2 hours, and dissolve in the mobile phase to make exactly
JP XVII Official Monographs / Amlodipine Besilate 417
50 mL. Pipet 25 mL of this solution, add exactly 10 mL of

the internal standard solution, add the mobile phase to make Amlodipine Besilate
100 mL, and use this solution as the standard solution.
Then, proceed as directed in the Assay under Amlexanox. アムロジピンベシル酸塩
Amount (mg) of amlexanox (C16H14N2O4)
＝ MS × QT/QS × V/200
MS: Amount (mg) of Amlexanox RS taken
Internal standard solution—A solution of 3-nitroaniline in
the mobile phase (1 in 500).
C20H25ClN2O5.C6H6O3S: 567.05
3-Ethyl 5-methyl (4RS )-2-[(2-aminoethoxy)methyl]-
medium, the dissolution rate in 45 minutes of Amlexanox
4-(2-chlorophenyl)-6-methyl-1,4-dihydropyridine-3,5-
dicarboxylate monobenzenesulfonate
Start the test with 1 tablet of Amlexanox Tablets, with-
[111470-99-6]
draw not less than 20 mL of the medium at the specified
minute after starting the test, and filter through a membrane
Amlodipine Besilate contains not less than 98.0z
filter with a pore size not exceeding 0.45 mm. Discard the
and not more than 102.0z of amlodipine besilate
first 10 mL of the filtrate, pipet V mL of the subsequent
(C20H25ClN2O5.C6H6O3S), calculated on the anhydrous
filtrate, add the dissolution medium to make exactly V? mL
basis.
so that each mL contains about 5.6 mg of amlexanox
(C16H14N2O4), and use this solution as the sample solution. Description Amlodipine Besilate occurs as a white to yel-
Separately, weigh accurately about 28 mg of Amlexanox RS, lowish white crystalline powder.
previously dried at 1059C for 2 hours, and dissolve in 2 mL It is freely soluble in methanol, sparingly soluble in
of dilute sodium hydroxide TS, add the dissolution medium ethanol (99.5), and slightly soluble in water.
to make exactly 50 mL. Pipet 1 mL of this solution, add the A solution of Amlodipine Besilate in methanol (1 in 100)
dissolution medium to make exactly 100 mL, and use this so- shows no optical rotation.
lution as the standard solution. Determine the absorbances, Melting point: about 1989C (with decomposition).
AT and AS, at 350 nm of the sample solution and standard
solution as directed under Ultraviolet-visible Spectropho-
solution of Amlodipine Besilate in 0.01 mol/L hydrochloric
tometry <2.24>.
acid-methanol TS (1 in 40,000) as directed under Ultraviolet-
Dissolution rate (z) with respect to the labeled amount visible Spectrophotometry <2.24>, and compare the spectrum
of amlexanox (C16H14N2O4) with the Reference Spectrum or the spectrum of a solution of
＝ MS × AT/AS × V?/V × 1/C × 18 Amlodipine Besilate RS prepared in the same manner as the
sample solution: both spectra exhibit similar intensities of
C: Labeled amount (mg) of amlexanox (C16H14N2O4) in 1
(2) Determine the infrared absorption spectrum of Am-
tablet
lodipine Besilate as directed in the potassium bromide disk
Assay Weigh accurately not less than 20 Amlexanox method under Infrared Spectrophotometry <2.25>, and com-
Tablets, and powder. Weigh accurately a portion of the pare the spectrum with the Reference Spectrum or the spec-
powder, equivalent to about 15 mg of amlexanox trum of Amlodipine Besilate RS: both spectra exhibit similar
(C16H14N2O4), add exactly 10 mL of the internal standard so- intensities of absorption at the same wave numbers.
lution, add 80 mL of the mobile phase, shake vigorously for (3) To 30 mg of Amlodipine Besilate add 0.1 g of sodium
5 minutes, and then add the mobile phase to make 100 mL. nitrate and 0.1 g of anhydrous sodium carbonate, mix, and
Centrifuge this solution, and use the supernatant liquid as gradually ignite. After cooling, dissolve the residue in 2 mL
the sample solution. Separately, weigh accurately about 30 of dilute hydrochloric acid and 10 mL of water, filter if nec-
mg of Amlexanox RS, previously dried at 1059C for 2 hours, essary, and add barium chloride TS: a white precipitate is
and dissolve in the mobile phase to make exactly 50 mL. formed.
Pipet 25 mL of this solution, add exactly 10 mL of the inter-
nal standard solution, add the mobile phase to make 100
Amlodipine Besilate according to Method 4, and perform
mL, and use this solution as the standard solution. Then,
proceed as directed in the Assay under Amlexanox.
Amount (mg) of amlexanox (C16H14N2O4) (2) Related substances—Dissolve 0.10 g of Amlodipine
＝ MS × QT/QS × 1/2 Besilate in 50 mL of a mixture of water and acetonitrile
(1:1), and use this solution as the sample solution. Pipet 1
mL of the sample solution, and add the mixture of water and
Internal standard solution—A solution of 3-nitroaniline in acetonitrile (1:1) to make exactly 100 mL. Pipet 3 mL of this
the mobile phase (1 in 500). solution, add the mixture of water and acetonitrile (1:1) to
tegration method: the area of the peak having the relative
418 Amlodipine Besilate Orally Disintegrating Tablets / Official Monographs JP XVII
retention time of 0.90 to amlodipine, obtained from the sam- QS, of the peak area of amlodipine to that of the internal
ple solution is not larger than the peak area of amlodipine standard.
obtained from the standard solution, and the area of the
Amount (mg) of amlodipine besilate
peak other than amlodipine, benzenesulfonic acid having the
(C20H25ClN2O5.C6H6O3S)
relative retention time of about 0.15 to amlodipine, and the
＝ M S × Q T / QS
peak mentioned above from the sample solution is not larger
than 1/3 times the peak area of amlodipine from the stand- MS: Amount (mg) of Amlodipine Besilate RS taken, cal-
ard solution. Furthermore, the total area of the peaks other culated on the anhydrous basis
than amlodipine and benzenesulfonic acid from the sample
Internal standard solution—A solution of isobutyl para-
solution is not larger than 2.7 times the peak area of amlodi-
hydroxybenzoate in the mobile phase (3 in 20,000).
pine from the standard solution.
length: 237 nm).
length: 237 nm).
259C.
359 C.
Mobile phase: A mixture of methanol and a solution of
Mobile phase A: A mixture of water and trifluoroacetic
potassium dihydrogen phosphate (41 in 10,000) (13:7).
acid (5000:1).
Flow rate: Adjust so that the retention time of amlodipine
Mobile phase B: A mixture of acetonitrile and trifluoroa-
is about 8 minutes.
cetic acid (5000:1).
ditions, amlodipine and the internal standard are eluted in
Time after injection Mobile phase A Mobile phase B this order with the resolution between these peaks being not
of sample (min) (volz) (volz) less than 3.
0 – 30 80 → 20 20 → 80
30 – 45 20 80
the peak area of amlodipine to that of the internal standard
Flow rate: 1.0 mL per minute. is not more than 1.0z.
retention time of amlodipine, beginning after the solvent Containers and storage Containers—Well-closed contain-
peak. ers.
System suitability— Storage—Light-resistant.
Test for required detectability: Pipet 1 mL of the standard
solution, and add a mixture of water and acetonitrile (1:1) to
make exactly 10 mL. Confirm that the peak area of amlodi- Amlodipine Besilate Orally
pine obtained with 10 mL of this solution is equivalent to 7 to
13z of that obtained with 10 mL of the standard solution.
Disintegrating Tablets
System performance: When the procedure is run with 10 アムロジピンベシル酸塩口腔内崩壊錠
ditions, the number of theoretical plates and the symmetry
factor of the peak of amlodipine are not less than 70,000 and Amlodipine Besilate Orally Disintegrating Tablets
not more than 1.5, respectively. contain not less than 95.0z and not more than
System repeatability: When the test is repeated 6 times 105.0z of the labeled amount of amlodipine besilate
with 10 mL of the standard solution under the above operat- (C20H25ClN2O5.C6H6O3S: 567.05).
ing conditions, the relative standard deviation of the peak Method of preparation Prepare as directed under Tablets,
area of amlodipine is not more than 2.0z. with Amlodipine Besilate.
Water <2.48> Not more than 0.5z (1 g, volumetric titra- Identification To an amount of powdered Amlodipine
tion, direct titration). Besilate Orally Disintegrating Tablets, equivalent to 7 mg of
Residue on ignition <2.44> Not more than 0.2z (1 g). Amlodipine Besilate, add 200 mL of 0.01 mol/L hydrochlo-
ric acid-methanol TS, treat with ultrasonic waves, and filter.
Assay Weigh accurately about 35 mg each of Amlodipine Determine the absorption spectrum of the filtrate as directed
Besilate and Amlodipine Besilate RS (separately determine under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-
the water <2.48> using the same manner as Amlodipine Besi- its a maximum between 358 nm and 362 nm.
late), dissolve them separately in the mobile phase to make
exactly 250 mL. Pipet 5 mL each of these solutions, add ex- Purity Related substances—Use the sample solution ob-
actly 5 mL each of the internal standard solution, add the tained in the Assay as the sample solution. Pipet 1 mL of the
mobile phase to make 25 mL, and use these solutions as the sample solution, add a mixture of methanol and the mobile
sample solution and standard solution. Perform the test with phase A (3:2) to make exactly 200 mL, and use this solution
20 mL of the sample solution and standard solution as di- as the standard solution. Perform the test with exactly 30 mL
rected under Liquid Chromatography <2.01> according to each of the sample solution and standard solution as directed
the following conditions, and calculate the ratios, QT and under Liquid Chromatography <2.01> according to the fol-
JP XVII Official Monographs / Amlodipine Besilate Orally Disintegrating Tablets 419
lowing conditions, and determine each peak area by the au- area of amlodipine is not more than 2.0z.
tomatic integration method: the area of the peak having the
relative retention time of about 0.45 to amlodipine obtained
amlodipine from the standard solution, the area of the peak
To 1 tablet of Amlodipine Besilate Orally Disintegrating
having the relative retention time of about 4.5 to amlodipine
Tablets add 4V/5 mL of a mixture of the mobile phase
from the sample solution is not larger than 1.8 times the
and methanol (1:1), disperse the particles with the aid of
peak area of amlodipine from the standard solution, and the
ultrasonic waves, add a mixture of the mobile phase and
area of the peak having the relative retention time of about
methanol (1:1) to make exactly V mL so that each mL of the
0.16 to amlodipine and the peaks other than mentioned
solution contains about 0.14 mg of amlodipine besilate
above from the sample solution is not larger than 2/5 times
(C20H25ClN2O5.C6H6O3S). Centrifuge this solution, and use
the peak area of amlodipine from the standard solution.
the supernatant liquid as the sample solution. Then, proceed
Furthermore, the total area of the peaks other than amlodi-
as directed in the Assay.
pine and the peak having the relative retention time of about
0.16 to amlodipine from the sample solution is not larger Amount (mg) of amlodipine besilate
than 2.8 times the peak area of amlodipine from the stand- (C20H25ClN2O5.C6H6O3S)
ard solution. For the areas of the peaks, having the relative ＝ MS × AT/AS × V × 1/250
retention time of about 0.45 and about 4.5 to amlodipine,
MS: Amount (mg) of Amlodipine Besilate RS taken, cal-
multiply their relative response factors, 2.0 and 1.9, respec-
culated on the anhydrous basis
tively.
Operating conditions— Disintegration Being specified separately when the drug is
Detector: An ultraviolet absorption photometer (wave- granted approval based on the Law.
length: 237 nm).
Dissolution Being specified separately when the drug is
granted approval based on the Law.
gel for liquid chromatography (5 mm in particle diameter). Assay Weigh accurately the mass of not less than 20 Am-
Column temperature: A constant temperature of about lodipine Besilate Orally Disintegrating Tablets, and powder
259 C. them. Weigh accurately a portion of this powder, equivalent
Mobile phase A: Dissolve 4.1 g of potassium dihydrogen to about 7 mg of amlodipine besilate (C20H25ClN2O5.
phosphate in 1000 mL of water, adjust to pH 6.0 with a solu- C6H6O3S), add 40 mL of a mixture of the mobile phase and
tion of 5.4 g of disodium hydrogen phosphate dodecahy- methanol (1:1), disperse the particles with the aid of ultra-
drate in 500 mL of water. To 500 mL of this solution add sonic waves, and add a mixture of the mobile phase and
500 mL of methanol. methanol (1:1) to make exactly 50 mL. Centrifuge this solu-
Mobile phase B: Dissolve 4.1 g of potassium dihydrogen tion, and use the supernatant liquid as the sample solution.
phosphate in 1000 mL of water, adjust to pH 6.0 with a solu- Separately, weigh accurately about 35 mg of Amlodipine
tion of 5.4 g of disodium hydrogen phosphate dodecahy- Besilate RS (separately, determine the water <2.48> in the
drate in 500 mL of water. To 50 mL of this solution add 950 same manner as Amlodipine Besilate), add 150 mL of a mix-
mL of methanol. ture of the mobile phase and methanol (1:1), dissolve with
Flowing of mobile phase: Control the gradient by mixing the aid of ultrasonic waves, then add a mixture of the mobile
the mobile phases A and B as directed in the following table. phase and methanol (1:1) to make exactly 250 mL, and use
Time after injection Mobile phase Mobile phase exactly 30 mL each of the sample solution and standard solu-
of sample (min) A (volz) B (volz) tion as directed under Liquid Chromatography <2.01>, and
determine the peak areas, AT and AS, of amlodipine in each
0 – 10 80 20 solution.
10 – 35 80 → 0 20 → 100
35 – 50 0 100 Amount (mg) of amlodipine besilate
＝ MS × AT/AS × 1/5
is about 10 minutes. MS: Amount (mg) of Amlodipine Besilate RS taken, cal-
Time span of measurement: About 5 times as long as the culated on the anhydrous basis
retention time of amlodipine.
length: 237 nm).
ard solution, and add a mixture of methanol and the mobile
phase A (3:2) to make exactly 50 mL. Confirm that the peak
area of amlodipine obtained with 30 mL of this solution is
equivalent to 14 to 26z of that obtained with 30 mL of the
standard solution.
259C.
phosphate in 1000 mL of water, adjust to pH 6.0 with a solu-
tion of 5.4 g of disodium hydrogen phosphate dodecahy-
factor of the peak of amlodipine are not less than 3000 and
drate in 500 mL of water. To 400 mL of this solution add
600 mL of methanol.
is about 10 minutes.
420 Amlodipine Besilate Tablets / Official Monographs JP XVII
System suitability— rately, determine the water <2.48> in the same manner as
System performance: When the procedure is run with 30 Amlodipine Besilate), and dissolve in the mobile phase to
mL of the standard solution under the above operating con- make exactly 250 mL. Pipet 5 mL of this solution, add ex-
ditions, the number of theoretical plates and the symmetry actly 5 mL of the internal standard solution, add the mobile
factor of the peak of amlodipine are not less than 3000 and phase to make 25 mL, and use this solution as the standard
not more than 2.0, respectively. solution. Perform the test with 20 mL each of the sample so-
System repeatability: When the test is repeated 6 times lution and standard solution as directed under Liquid Chro-
with 30 mL of the standard solution under the above operat- matography <2.01> according to the following conditions,
ing conditions, the relative standard deviation of the peak and calculate the ratios, QT and QS, of the peak area of am-
area of amlodipine is not more than 1.0z. lodipine to that of the internal standard.
Containers and storage Containers—Tight containers. Amount (mg) of amlodipine besilate
＝ MS × QT/QS × 1/50
Amlodipine Besilate Tablets MS: Amount (mg) of Amlodipine Besilate RS taken, cal-
アムロジピンベシル酸塩錠
Amlodipine Besilate Tablets contain not less than
amount of amlodipine besilate (C20H25ClN2O5.
length: 237 nm).
C6H6O3S: 567.05).
Method of preparation Prepare as directed under Tablets, ter and 15 cm in length, packed with octadecylsilanized silica
with Amlodipine Besilate. gel for liquid chromatography (5 mm in particle diameter).
Identification To a quantity of powdered Amlodipine Besi-
259C.
late Tablets, equivalent to 2.5 mg of Amlodipine Besilate,
Mobile phase: A mixture of methanol and potassium dihy-
add 100 mL of 0.01 mol/L hydrochloric acid-methanol TS,
drogen phosphate (41 in 10,000) (13:7).
shake vigorously, and filter. Determine the absorption spec-
trum of the filtrate as directed under Ultraviolet-visible Spec-
is about 8 minutes.
trophotometry <2.24>: it exhibits maxima between 235 nm
and 239 nm, and between 358 nm and 362 nm.
Uniformity of dosage units <6.02> Perform the test accord- mL of the standard solution under the above operating con-
ing to the following method: it meets the requirement of the ditions, amlodipine and the internal standard are eluted in
Content uniformity test. this order with the resolution between these peaks being not
To 1 tablet of Amlodipine Besilate Tablets add 10 mL of less than 5.
water to disintegrate, disperse with the aid of ultrasonic System repeatability: When the test is repeated 6 times
waves with occasional shaking, add the mobile phase to with 20 mL of the standard solution under the above operat-
make exactly V mL so that each mL contains about 69 mg of ing conditions, the relative standard deviation of the ratios
amlodipine besilate (C20H25ClN2O5.C6H6O3S), and shake for of the peak area of amlodipine to that of the internal stand-
60 minutes. Centrifuge this solution, pipet 10 mL of the su- ard is not more than 1.0z.
pernatant liquid, add exactly 5 mL of the internal standard
solution, add the mobile phase to make 25 mL, and use this
ers.
solution as the sample solution. Proceed as directed in the
Assay.
Amount (mg) of amlodipine besilate Ammonia Water
＝ MS × QT/QS × V/500 アンモニア水
MS: Amount (mg) of Amlodipine Besilate RS taken, cal-
culated on the anhydrous basis Ammonia Water contains not less than 9.5 w/vz
and not more than 10.5 w/vz of ammonia (NH3:
17.03).
Description Ammonia Water occurs as a clear, colorless
liquid, having a very pungent, characteristic odor.
It is alkaline.
Assay To 20 Amlodipine Besilate Tablets add 100 mL of Specific gravity d 20
20: 0.95 – 0.96
water to disintegrate, disperse with the aid of ultrasonic
Identification (1) Hold a glass rod moistened with hydro-
waves with occasional shaking, add the mobile phase to
chloric acid near the surface of Ammonia Water: dense
make exactly 1000 mL, and shake for 60 minutes. Centrifuge
white fumes are produced.
this solution, pipet a volume of the supernatant liquid,
(2) Hold moistened red litmus paper near the surface of
equivalent to about 0.7 mg of amlodipine besilate
Ammonia Water: it turns blue.
(C20H25ClN2O5.C6H6O3S), add exactly 5 mL of the internal
standard solution, add the mobile phase to make 25 mL, and Purity (1) Residue on evaporation—Evaporate 10.0 mL
use this solution as the sample solution. Separately, weigh of Ammonia Water to dryness, and dry the residue at 1059C
accurately about 35 mg of Amlodipine Besilate RS (sepa- for 1 hour: the mass of the residue is not more than 2.0 mg.
JP XVII Official Monographs / Amosulalol Hydrochloride 421
(2) Heavy metals <1.07>—Evaporate 5.0 mL of Ammo- dry at 1059C for 30 minutes: the crystals so obtained melt
nia Water to dryness on a water bath, add 1 mL of dilute <2.60> between 1689C and 1739C or between 1509C and
hydrochloric acid to the residue, and evaporate to dryness. 1549C.
Dissolve the residue in 2 mL of dilute acetic acid, add water
the test solution. Prepare the control solution with 2.5 mL of Purity (1) Clarity and color of solution—Dissolve 0.5 g
Standard Lead Solution, 2 mL of dilute acetic acid and water of Amobarbital in 5 mL of sodium hydroxide TS: the solu-
to make 50 mL (not more than 5 ppm). tion is clear and colorless.
(3) Potassium permanganate-reducing substances—To (2) Chloride <1.03>—Dissolve 0.30 g of Amobarbital in
10.0 mL of Ammonia Water add 40 mL of dilute sulfuric 20 mL of acetone, and add 6 mL of dilute nitric acid and
acid while cooling, and add 0.10 mL of 0.02 mol/L potas- water to make 50 mL. Perform the test using this solution as
sium permanganate VS: the red color of the potassium per- the test solution. Prepare the control solution as follows:
manganate does not disappear within 10 minutes. take 0.30 mL of 0.01 mol/L hydrochloric acid VS, 20 mL of
acetone and 6 mL of dilute nitric acid, and add water to
Assay Measure exactly 5 mL of Ammonia Water, add 25
mL of water, and titrate <2.50> with 0.5 mol/L sulfuric acid
(3) Sulfate <1.14>—Dissolve 0.40 g of Amobarbital in 20
VS (indicator: 2 drops of methyl red TS).
mL of acetone, and add 1 mL of dilute hydrochloric acid
Each mL of 0.5 mol/L sulfuric acid VS and water to make 50 mL. Perform the test using this solu-
＝ 17.03 mg of NH3 tion as the test solution. Prepare the control solution as
follows: take 0.40 mL of 0.005 mol/L sulfuric acid VS, 20
mL of acetone, and 1 mL of dilute hydrochloric acid, and
Storage—Not exceeding 309C.
add water to make 50 mL (not more than 0.048z).
(4) Heavy metals <1.07>—Proceed with 1.0 g of Amobar-
bital according to Method 2, and perform the test. Prepare
Amobarbital the control solution with 2.0 mL of Standard Lead Solution
アモバルビタール
(5) Readily carbonizable substances <1.15>—Perform the
test with 0.5 g of Amobarbital. The solution is not more
colored than Matching Fluid A.
4 hours).
C11H18N2O3: 226.27
5-Ethyl-5-(3-methylbutyl)pyrimidine- Assay Weigh accurately about 0.5 g of Amobarbital, previ-
2,4,6(1H,3H,5H )-trione ously dried, and dissolve in 5 mL of ethanol (95) and 50 mL
[57-43-2] of chloroform. Titrate <2.50> with 0.1 mol/L potassium
hydroxide-ethanol VS until the color of the solution changes
Amobarbital, when dried, contains not less than from yellow through light blue to purple (indicator: 1 mL of
99.0z of amobarbital (C11H18N2O3). alizarin yellow GG-thymolphthalein TS). Perform a blank
Description Amobarbital occurs as white, crystals or crys-
talline powder. It is odorless, and has a slightly bitter taste. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS
It is freely soluble in ethanol (95), in acetone and in diethyl ＝ 22.63 mg of C11H18N2O3
ether, sparingly soluble in chloroform, and practically in-
soluble in water.
ers.
It dissolves in sodium hydroxide TS and in sodium car-
bonate TS.
The pH of a saturated solution of Amobarbital is between
5.0 and 5.6. Amosulalol Hydrochloride
Identification (1) Boil 0.2 g of Amobarbital with 10 mL アモスラロール塩酸塩
of sodium hydroxide TS: the gas evolved changes moistened
red litmus paper to blue.
(2) Dissolve 0.05 g of Amobarbital in 2 to 3 drops of am-
monia-ammonium chloride buffer solution (pH 10.7) and
5 mL of diluted pyridine (1 in 10). Add 5 mL of chloroform
and 0.3 mL of copper (II) sulfate TS to the solution: a red-
purple precipitate is produced in the aqueous layer. Shake
the mixture: a red-purple color is produced in the chlo- C18H24N2O5S.HCl: 416.92
roform layer. 5-((1RS )-1-Hydroxy-2-{[2-(2-
(3) To 0.4 g of Amobarbital add 0.1 g of anhydrous methoxyphenoxy)ethyl]amino}ethyl)-
sodium carbonate and 4 mL of water, shake, and add a solu- 2-methylbenzenesulfonamide monohydrochloride
tion of 0.3 g of 4-nitrobenzyl chloride in 7 mL of ethanol [70958-86-0]
(95). Heat the mixture on a water bath for 30 minutes under
a reflux condenser, and allow to stand for 1 hour. Filter the Amosulalol Hydrochloride contains not less than
crystals produced, wash with 7 mL of sodium hydroxide TS 98.5z and not more than 101.0z of amosulalol hy-
and a small portion of water, recrystallize from ethanol, and drochloride (C18H24N2O5S.HCl), calculated on the an-
422 Amosulalol Hydrochloride Tablets / Official Monographs JP XVII
hydrous basis. peak.

Description Amosulalol Hydrochloride occurs as white
crystals or a white crystalline powder. It has a bitter taste.
solution, and add the mobile phase to make exactly 10 mL.
It is very soluble in formic acid, freely soluble in metha-
Confirm that the peak area of amosulalol obtained with
nol, and sparingly soluble in water and in ethanol (99.5).
10 mL of this solution is equivalent to 7 to 13z of that ob-
It is hygroscopic.
tained with 10 mL of the standard solution.
A solution of Amosulalol Hydrochloride in methanol (1 in
100) shows no optical rotation.
Identification (1) Determine the absorption spectrum of ditions, the number of theoretical plates and the symmetry
a solution of Amosulalol Hydrochloride (1 in 20,000) as factor of the peak of amosulalol are not less than 4000 and
directed under Ultraviolet-visible Spectrophotometry <2.24>, not more than 1.7, respectively.
and compare the spectrum with the Reference Spectrum: System repeatability: When the test is repeated 6 times
both spectra exhibit similar intensities of absorption at the with 10 mL of the standard solution under the above operat-
same wavelengths. ing conditions, the relative standard deviation of the peak
(2) Determine the infrared absorption spectrum of area of amosulalol is not more than 1.0z.
Amosulalol Hydrochloride as directed in the potassium chlo-
Water <2.48> Not more than 4.0z (1 g, volumetric titra-
ride disk method under Infrared Spectrophotometry <2.25>,
both spectra exhibit similar intensities of absorption at the Residue on ignition <2.44> Not more than 0.1z (1 g).
same wave numbers.
Assay Weigh accurately about 0.6 g of Amosulalol Hydro-
(3) A solution of Amosulalol Hydrochloride (1 in 100)
chloride, dissolve in 3 mL of formic acid, add 80 mL of a
mixture of acetic acid (100) and acetic anhydride (3:2), and
Melting point <2.60> 158 – 1629C titrate <2.50> within 5 minutes with 0.1 mol/L perchloric
Purity (1) Heavy metals <1.07>—Place 1.0 g of
nation using the same procedure, and make any necessary
Amosulalol Hydrochloride in a porcelain crucible, add 1.5
correction.
mL of sulfuric acid, cover loosely, and heat gently to car-
bonize. After cooling, add 2 mL of nitric acid, heat carefully Each mL of 0.1 mol/L perchloric acid VS
until white fumes no longer are evolved, and then heat inten- ＝ 41.69 mg of C18H24N2O5S.HCl
sely to 500 – 6009C to incinerate. After cooling, add 2 mL of
hydrochloric acid, proceed according to Method 2, and per-
form the test. The control solution, processed in the same
manner as the test solution using the same amounts of rea-
gents, is prepared by combining 2.0 mL of Standard Lead Amosulalol Hydrochloride Tablets
Solution and water to make 50 mL (not more than 20 ppm).
アモスラロール塩酸塩錠
(2) Related substances—Dissolve 0.10 g of Amosulalol
solution as the sample solution. Pipet 1 mL of the sample so- Amosulalol Hydrochloride Tablets contain not
lution, add the mobile phase to make exactly 200 mL, and less than 95.0z and not more than 105.0z of
use this solution as the standard solution. Perform the test the labeled amount of amosulalol hydrochloride
with exactly 10 mL each of the sample solution and standard (C18H24N2O5S.HCl: 416.92).
according to the following conditions, and determine each
with Amosulalol Hydrochloride.
peak area of both solutions by the automatic integration
method: the total area of the peaks other than amosulalol Identification To a quantity of powdered Amosulalol
obtained from the sample solution is not larger than 2/5 Hydrochloride Tablets, equivalent to 50 mg of Amosulalol
times the peak area of amosulalol obtained from the stand- Hydrochloride, add 25 mL of 0.1 mol/L hydrochloric acid
ard solution. TS, shake well, and then centrifuge. To 2.5 mL of the super-
Operating conditions— natant liquid add water to make 100 mL. Determine the ab-
Detector: An ultraviolet absorption photometer (wave- sorption spectrum of this solution as directed under Ultravi-
length: 272 nm). olet-visible Spectrophotometry <2.24>: it exhibits a maxi-
Column: A stainless steel column 4.6 mm in inside diame- mum between 270 nm and 274 nm, and a shoulder between
ter and 15 cm in length, packed with octadecylsilanized silica 275 nm and 281 nm.
309 C.
Take 1 tablet of Amosulalol Hydrochloride Tablets, disin-
phosphate in water to make 1000 mL, and adjust to pH 5.7
tegrate by adding 2 mL of 0.1 mol/L hydrochloric acid TS,
by adding a solution prepared by dissolving 3.58 g of disodi-
add 15 mL of methanol, and shake well. Add methanol to
um hydrogen phosphate dodecahydrate in water to make
1000 mL. To 670 mL of this solution add 330 mL of aceto-
of amosulalol hydrochloride (C18H24N2O5S.HCl), and centri-
nitrile.
fuge. Pipet 5 mL of the supernatant liquid, add exactly 2 mL
Flow rate: Adjust so that the retention time of amosulalol
of the internal standard solution and the mobile phase to
is about 7 minutes.
Separately, weigh accurately about 20 mg of amosulalol
retention time of amosulalol, beginning after the solvent
JP XVII Official Monographs / Amosulalol Hydrochloride Tablets 423
hydrochloride for assay (separately determine the water ditions, the number of theoretical plates and the symmetry
<2.48> in the same manner as Amosulalol Hydrochloride), factor of the peak of amosulalol are not less than 4000 and
and dissolve in methanol to make exactly 50 mL. Pipet 5 mL not more than 1.7, respectively.
of this solution, add exactly 2 mL of the internal standard System repeatability: When the test is repeated 6 times
solution, add the mobile phase to make 20 mL, and use this with 50 mL of the standard solution under the above operat-
solution as the standard solution. Then, proceed as directed ing conditions, the relative standard deviation of the peak
in the Assay. area of amosulalol is not more than 1.0z.
Amount (mg) of amosulalol hydrochloride Assay Take 10 Amosulalol Hydrochloride Tablets, add
(C18H24N2O5S.HCl) 20 mL of 0.1 mol/L hydrochloric acid TS, and shake well to
＝ MS × QT/QS × V/50 disintegrate. Add 120 mL of methanol, again shake well,
add methanol to make exactly 200 mL, and then centrifuge.
MS: Amount (mg) of amosulalol hydrochloride for assay
Pipet a volume of supernatant liquid corresponding to about
taken, calculated on the anhydrous basis
5 mg of amosulalol hydrochloride (C18H24N2O5S.HCl), add
Internal standard solution—A solution of ethyl para- exactly 5 mL of the internal standard solution, add the mo-
hydroxybenzoate in methanol (1 in 6250). bile phase to make 50 mL, and use this solution as the sam-
amosulalol hydrochloride for assay (separately determine the
water <2.48> in the same manner as Amosulalol Hydrochlo-
ride), and dissolve in methanol to make exactly 25 mL. Pipet
in 30 minutes of Amosulalol Hydrochloride Tablets is not
5 mL of this solution, add exactly 5 mL of the internal stand-
less than 75z.
ard solution, add the mobile phase to make 50 mL, and use
Start the test with 1 tablet of Amosulalol Hydrochloride
directed under Liquid Chromatography <2.01> according to
the following conditions, and calculate the ratios, QT and
QS, of the peak area of amosulalol to that of the internal
quent filtrate, add water to make exactly V? mL so that each
standard.
mL contains about 5.5 mg of amosulalol hydrochloride
(C18H24N2O5S.HCl), and use this solution as the sample Amount (mg) of amosulalol hydrochloride
solution. Separately, weigh accurately about 22 mg of (C18H24N2O5S.HCl)
amosulalol hydrochloride for assay (separately determine the ＝ MS × QT/QS × 1/5
water <2.48> in the same manner as Amosulalol Hydrochlo-
MS: Amount (mg) of amosulalol hydrochloride for assay
ride), and dissolve in water to make exactly 200 mL. Pipet 5
mL of this solution, add water to make exactly 100 mL, and
use this solution as the standard solution. Perform the test Internal standard solution—A solution of ethyl para-
with exactly 50 mL each of the sample solution and standard hydroxybenzoate in methanol (1 in 6250).
solution as directed under Liquid Chromatography <2.01> Operating conditions—
according to the following conditions, and determine the Detector: An ultraviolet absorption photometer (wave-
amosulalol peak areas, AT and AS, in each solution. length: 272 nm).
of amosulalol hydrochloride (C18H24N2O5S.HCl)
＝ MS × AT/AS × V?/V × 1/C × 45/2
MS: Amount (mg) of amosulalol hydrochloride for assay 259C.
taken, calculated on the anhydrous basis Mobile phase: A mixture of diluted acetic acid (100) (1 in
C: Labeled amount (mg) of amosulalol hydrochloride 25), acetonitrile and a solution of ammonium acetate (1 in
(C18H24N2O5S.HCl) in 1 tablet 250) (5:3:2).
is about 4 minutes.
length: 272 nm).
ditions, amosulalol and the internal standard are eluted in
less than 7.
309 C.
by adding a soluion prepared by dissolving 3.58 g of disodi-
the peak area of amosulalol to that of the internal standard
um hydrogen phosphate dodecahydrate in water to make
nitrile. Containers and storage Containers—Tight containers.
is about 5 minutes.
424 Amoxapine / Official Monographs JP XVII
Amoxapine <2.50> with 0.1 mol/L perchloric acid VS until the color of
the solution changes from purple through blue to greenish
アモキサピン blue (indicator: 2 drops of crystal violet TS). Perform a
blank determination, and make any necessary correction.
＝ 15.69 mg of C17H16ClN3O
Amoxicillin Hydrate
C17H16ClN3O: 313.78
2-Chloro-11-(piperazin-1-yl)dibenzo[b, f ][1,4]oxazepine アモキシシリン水和物
[14028-44-5]
Amoxapine, when dried, contains not less than

98.5z of amoxapine (C17H16ClN3O).
Description Amoxapine occurs as white to light yellowish
white, crystals or crystalline powder.
It is freely soluble in acetic acid (100), slightly soluble in C16H19N3O5S.3H2O: 419.45
ethanol (95) and in diethyl ether, and practically insoluble in (2S,5R,6R)-6-[(2R)-2-Amino-2-(4-hydroxyphenyl)-
water. acetylamino]-3,3-dimethyl-7-oxo-4-thia-1-
azabicyclo[3.2.0]heptane-2-carboxylic acid trihydrate
[61336-70-7]
solution of Amoxapine in 0.1 mol/L hydrochloric acid TS
(1 in 50,000) as directed under Ultraviolet-visible Spectro-
Amoxicillin Hydrate contains not less than 950 mg
(potency) and not more than 1010 mg (potency) per
mg, calculated on the anhydrous basis. The potency of
absorption as the same wavelengths.
Amoxicillin Hydrate is expressed as mass (potency) of
amoxicillin (C16H19N3O5S: 365.40).
Amoxapine, previously dried, as directed in the potassium
bromide disk method under Infrared Spectrophotometry Description Amoxicillin Hydrate occurs as white to light
<2.25>, and compare the spectrum with the Reference Spec- yellowish white, crystals or crystalline powder.
trum: both spectra exhibit similar intensities of absorption at It is slightly soluble in water and in methanol, and very
the same wave numbers. slightly soluble in ethanol (95).
(3) Perform the test with Amoxapine as directed under
Identification Determine the infrared absorption spectrum
Flame Coloration Test <1.04> (2): a green color appears.
of Amoxicillin Hydrate as directed in the potassium bromide
Melting point <2.60> 178 – 1829C disk method under Infrared Spectrophotometry <2.25>, and
compare the spectrum with the Reference Spectrum or the
spectrum of Amoxicillin RS: both spectra exhibit similar in-
Amoxapine according to Method 2, and perform the test.
tensities of absorption at the same wave numbers.
Solution (not more than 15 ppm). Optical rotation <2.49> [a]20
D : ＋290 – ＋3159(0.1 g calcu-
(2) Related substances—Dissolve 0.5 g of Amoxapine in lated on the anhydrous basis, water, 100 mL, 100 mm).
10 mL of a mixture of ethanol (95) and acetic acid (100)
Purity (1) Heavy metals <1.07>—To 1.0 g of Amoxicillin
Hydrate add 2 mL of a solution of magnesium sulfate hepta-
mL of the sample solution, and add a mixture of ethanol
hydrate (1 in 4), mix, and heat on a water bath to dryness.
(95) and acetic acid (100) (9:1) to make exactly 10 mL. Pipet
Carbonize the residue by gently heating. After cooling, add 1
1 mL of this solution, add a mixture of ethanol (95) and
mL of sulfuric acid, heat carefully, then heat at 500 – 6009C
acetic acid (100) (9:1) to make exactly 20 mL, and use this
to incinerate. After cooling, add 1 mL of hydrochloric acid
solution as the standard solution. Perform the test with these
to the residue, and heat on a water bath to dryness. Then
add 10 mL of water to the residue, and heat on a water bath
to dissolve. After cooling, add ammonia TS to adjust the pH
solution on a plate of silica gel with fluorescent indicator for
to 3 – 4, and add 2 mL of dilute acetic acid. If necessary,
filter, wash the residue on the filter with 10 mL of water,
of ethanol (95) and acetic acid (100) (9:1) to a distance of
transfer the filtrate and washings into a Nessler tube, add
about 10 cm, and air-dry the plate. Examine under ultravio-
water to make 50 mL, and use this solution as the test solu-
let light (main wavelength: 254 nm): the spots other than the
tion. Prepare the control solution as follows: To 2.0 mL of
principal spot from the sample solution are not more intense
Standard Lead Solution add 2 mL of a solution of magne-
sium sulfate heptahydrate (1 in 4), then proceed in the same
Loss on drying <2.41> Not more than 0.4z (1 g, in vacu- manner as for preparation of the test solution (not more
um, 609C, 3 hours). than 20 ppm).
of Amoxicillin Hydrate according to Method 4, and perform
Assay Weigh accurately about 0.3 g of Amoxapine, previ- the test (not more than 2 ppm).
JP XVII Official Monographs / Amoxicillin Capsules 425
(3) Related substances—Dissolve 0.10 g of Amoxicillin gel for liquid chromatography (5 mm in particle diameter).
Hydrate in 50 mL of a solution of boric acid (1 in 200), and Column temperature: A constant temperature of about
use this solution as the sample solution. Pipet 1 mL of the 259C.
sample solution, add a solution of boric acid (1 in 200) to Mobile phase: Dissolve 1.361 g of sodium acetate trihy-
make exactly 100 mL, and use this solution as the standard drate in 750 mL of water, adjust to pH 4.5 with acetic acid
solution. Perform the test with exactly 10 mL each of the (31), and add water to make 1000 mL. To 950 mL of this so-
sample solution and standard solution as directed under Liq- lution add 50 mL of methanol.
uid Chromatography <2.01> according to the following con- Flow rate: Adjust so that the retention time of amoxicillin
ditions. Determine each peak area of both solutions by the is about 8 minutes.
automatic integration method: the area of each peak other System suitability—
than amoxicillin obtained from the sample solution is not System performance: When the procedure is run with
larger than the peak area of amoxicillin obtained from the 10 mL of the standard solution under the above operating
standard solution. conditions, the number of theoretical plates of the peak of
Operating conditions— amoxicillin is not less than 2500.
Detector: An ultraviolet absorption photometer (wave- System repeatability: When the test is repeated 6 times
length: 254 nm). with 10 mL of the standard solution under the above operat-
Column: A stainless steel column 4 mm in inside diameter ing conditions, the relative standard deviation of the peak
and 30 cm in length, packed with octadecylsilanized silica gel area of amoxicillin is not more than 1.0z.
259 C.
Mobile phase: Dissolve 1.36 g of sodium acetate trihydrate
in 750 mL of water, adjust to pH 4.5 with acetic acid (31), Amoxicillin Capsules
アモキシシリンカプセル
add 50 mL of methanol.
Flow rate: Adjust so that the retention time of amoxicillin
is about 8 minutes. Amoxicillin Capsules contain not less than 92.0z
Time span of measurement: About 4 times as long as the and not more than 105.0z of the labeled potency of
retention time of amoxicillin. Amoxicillin (C16H19N3O5S: 365.40).
Method of preparation Prepare as directed under Cap-
sules, with Amoxicillin Hydrate.
standard solution add a solution of boric acid (1 in 200) to
make exactly 10 mL. Confirm that the peak area of amox- Identification Take out the contents of Amoxicillin Cap-
icillin obtained from 10 mL of this solution is equivalent to 7 sules, to a quantity of the contents, equivalent to 8 mg
to 13z of that obtained from 10 mL of the standard solu- (potency) of Amoxicillin Hydrate, add 2 mL of 0.01 mol/L
tion. hydrochloric acid TS, shake for 30 minutes, filter, and use
System performance: When the procedure is run with 10 the filtrate as the sample solution. Separately, dissolve an
mL of the standard solution under the above operating con- amount equivalent to 8 mg (potency) of Amoxicillin RS in 2
ditions, the number of theoretical plates and the symmetry mL of 0.01 mol/L hydrochloric acid TS, and use this solu-
factor of the peak of amoxicillin are not less than 2500 and tion as the standard solution. Perform the test with these so-
not nore than 1.5, respectively. lutions as directed under Thin-layer Chromatography <2.03>.
System repeatability: When the test is repeated 6 times Spot 5 mL each of the sample solution and standard solution
with 10 mL of the standard solution under the above operat- on a plate of silica gel for thin-layer chromatography. De-
ing conditions, the relative standard deviation of the peak velop the plate with a mixture of tetrahydrofuran, water and
area of amoxicillin is not more than 1.0z. formic acid (50:5:2) to a distance of about 10 cm, and air-
dry the plate. Spray evenly a solution of ninhydrin in ethanol
Water <2.48> Not less than 11.0z and not more than
(95) (1 in 20) on the plate, and heat the plate at 1109C for 15
15.0z (0.1 g, volumetric titration, direct titration).
minutes: the principal spot obtained from the sample solu-
Assay Weigh accurately an amount of Amoxicillin Hydrate tion and the spot obtained from the standard solution show
and Amoxicillin RS, equivalent to about 30 mg (potency), a red-purple color and the same R f value.
dissolve each in a solution of boric acid (1 in 200) to make
Purity Related substances—Take out the contents of
exactly 100 mL, and use these solutions as the sample solu-
Amoxicillin Capsules, to a quantity of the contents, equiva-
tion and standard solution. Perform the test with exactly
lent to 0.1 g (potency) of Amoxicillin Hydrate, add 30 mL of
a solution of boric acid (1 in 200), shake for 15 minutes, and
add a solution of boric acid (1 in 200) to make 50 mL. Cen-
the following conditions, and calculate the peak areas, AT
trifuge this solution, and use the supernatant liquid as the
and AS, of amoxicillin in each solution.
sample solution. Pipet 1 mL of the sample solution, add a
Amount [ mg (potency)] of amoxicillin (C16H19N3O5S) solution of boric acid (1 in 200) to make exactly 100 mL, and
＝ MS × AT/AS × 1000 use this solution as the standard solution. Perform the test
MS: Amount [mg (potency)] of Amoxicillin RS taken
Operating conditions— according to the following conditions. Determine each peak
Detector: An ultraviolet absorption photometer (wave- area of both solutions by the automatic integration method:
length: 230 nm). the area of each peak other than amoxicillin obtained from
Column: A stainless steel column 4.6 mm in inside diame- the sample solution is not larger than the peak area of
ter and 15 cm in length, packed with octadecylsilanized silica amoxicillin obtained from the standard solution.
426 Amphotericin B / Official Monographs JP XVII
Operating conditions— weigh accurately an amount equivalent to about 20 mg (po-
Proceed as directed in the operating conditions in the tency) of Amoxicillin RS, dissolve in water to make exactly
Purity (3) under Amoxicillin Hydrate. 20 mL, and use this solution as the standard solution. Per-
System suitability— form the test with exactly 10 mL each of the sample solution
Test for required detectability and system repeatability: and standard solution as directed under Liquid Chromatog-
Proceed as directed in the system suitability in the Purity (3) raphy <2.01> according to the following conditions, and
under Amoxicillin Hydrate. determine the peak areas, AT and AS, of amoxicillin in each
System performance: When the procedure is run with 10 solution.
Amount [mg (potency)] of amoxicillin (C16H19N3O5S)
＝ MS × AT/AS × 5
factor of the peak of amoxicillin is not less than 2500 and
not more than 1.5, respectively. MS: Amount [mg (potency)] of Amoxicillin RS taken
Water <2.48> Not more than 15.0z (0.1 g, volumetric Operating conditions—
titration, direct titration). Column temperature, mobile phase, and flow rate: Pro-
ceed as directed in the operating conditions in the Assay
under Amoxicillin Hydrate.
Dissolution <6.10> When the test is performed at 100 revo- length: 254 nm).
lutions per minute according to the Paddle method using the Column: A stainless steel column 4 mm in inside diameter
sinker, using 900 mL of water as the dissolution medium, the and 30 cm in length, packed with octadecylsilanized silica gel
dissolution rate in 60 minutes of Amoxicillin Capsules is not for liquid chromatography (10 mm in particle diameter).
less than 75z. System suitability—
Start the test with 1 capsule of Amoxicillin Capsules, with- System performance: When the procedure is run with 10
draw not less than 20 mL of the medium at the specified mL of the standard solution under the above operating con-
minute after starting the test, and filter through a membrane ditions, the number of theoretical plates and the symmetry
filter with a pore size not exceeding 0.45 mm. Discard the factor of the peak of amoxicillin are not less than 2500 and
first 10 mL of the filtrate, pipet V mL of the subsequent not more than 1.5, respectively.
filtrate, add water to make exactly V? mL so that each mL System repeatability: When the test is repeated 6 times
contains about 56 mg (potency) of Amoxicillin Hydrate, and with 10 mL of the standard solution under the above operat-
use this solution as the sample solution. Separately, weigh ing conditions, the relative standard deviation of peak areas
accurately an amount equivalent to about 28 mg (potency) of of amoxicillin is not more than 1.0z.
Amoxicillin RS, dissolve in water to make exactly 100 mL.
the test with exactly 50 mL each of the sample solution and
standard solution as directed under Liquid Chromatography Amphotericin B
<2.01> according to the following conditions, and determine
アムホテリシン B
the peak areas, AT and AS, of amoxicillin in each solution.
of amoxicillin (C16H19N3O5S)
＝ MS × AT/AS × V?/V × 1/C × 180
MS: Amount [mg (potency)] of Amoxicillin RS taken
C: Labeled amount [mg (potency)] of amoxicillin
(C16H19N3O5S) in 1 capsule
Assay under Amoxicillin Hydrate.
System performance: When the procedure is run with 50 C47H73NO17: 924.08
mL of the standard solution under the above operating con- (1R,3S,5R,6R,9R,11R,15S,16R,17R,18S,19E,21E,
ditions, the number of theoretical plates and the symmetry 23E,25E,27E,29E,31E,33R,35S,36R,37S )-33-(3-
factor of the peak of amoxicillin are not less than 2500 and Amino-3,6-dideoxy-b-D-mannopyranosyloxy)-
not more than 2.0, respectively. 1,3,5,6,9,11,17,37-octahydroxy-15,16,18-trimethyl-13-oxo-
System repeatability: When the test is repeated 6 times 14,39-dioxabicyclo[33.3.1]nonatriaconta-
with 50 mL of the standard solution under the above operat- 19,21,23,25,27,29,31-heptaene-36-carboxylic acid
ing conditions, the relative standard deviation of the peak [1397-89-3]
area of amoxicillin is not more than 1.5z.
Amphotericin B is a polyene macrolide substance
having antifungal activity produced by the growth of
Amoxicillin Capsules, take out the contents, and weigh accu-
Streptomyces nodosus.
rately the mass of the emptied shells. Weigh accurately an
It contains not less than 840 mg (potency) per mg,
amount equivalent to about 0.1 g (potency) of Amoxicillin
calculated on the dried basis. The potency of Am-
Hydrate, add 70 mL of water, shake for 15 minutes, and add
photericin B is expressed as mass (potency) of am-
water to make exactly 100 mL. Centrifuge this solution, and
photericin B (C47H73NO17).
JP XVII Official Monographs / Amphotericin B for Injection 427
Description Amphotericin B occurs as a yellow to orange Weigh accurately an amount of Amphotericin B RS equiva-
powder. lent to about 20 mg (potency), dissolve in dimethylsulfoxide
It is freely soluble in dimethylsulfoxide and practically to make exactly 20 mL, and use this solution as the standard
insoluble in water and in ethanol (95). stock solution. Keep the standard stock solution at 59 C or
below and use within 24 hours. Take exactly a suitable
Identification (1) Dissolve 5 mg of Amphotericin B in 10
amount of the standard stock solution before use, and add
mL of dimethylsulfoxide. To 1 mL of this solution add 5 mL
dimethylsulfoxide to make solutions so that each mL con-
of phosphoric acid: a blue color develops between the two
tains 200 mg (potency) and 50 mg (potency). Pipet 1 mL each
layers, and the solution becomes blue by shaking. After ad-
of these solutions, add 0.2 mol/L phosphate buffer solution
dition of 15 mL of water it becomes yellow to light yellow-
(pH 10.5) to make exactly 20 mL, and use these solutions as
brown by shaking.
the high concentration standard solution and low concentra-
(2) Dissolve 25 mg of Amphotericin B in 5 mL of
tion standard solution, respectively.
dimethylsulfoxide, and add methanol to make 50 mL. To 1
(v) Sample solution—Use light-resistant vessels. Weigh
mL of this solution add methanol to make 50 mL. Determine
accurately an amount of Amphotericin B equivalent to about
the absorption spectrum of this solution as directed under
20 mg (potency), dissolve in dimethylsulfoxide to make
exactly 20 mL, and use this solution as the sample stock so-
lution. Take exactly a suitable amount of the sample stock
of a solution of Amphotericin B RS prepared in the same
solution, add dimethylsulfoxide to make solutions so that
manner as the sample solution: both spectra exhibit similar
each mL contains 200 mg (potency) and 50 mg (potency).
intensities of absorption at the same wavelengths.
Pipet 1 mL each of these solutions, add 0.2 mol/L phos-
Purity Amphotericin A—Weigh accurately about 50 mg phate buffer solution (pH 10.5) to make exactly 20 mL, and
each of Amphotericin B and Amphotericin B RS, add ex- use these solutions as the high concentration sample solution
actly 10 mL each of dimethylsulfoxide to dissolve, and add and low concentration sample solution, respectively.
methanol to make exactly 50 mL. Pipet 4 mL each of these
solutions, add methanol to make exactly 50 mL, and use
Storage—Light-resistant, and in a cold place.
these solutions as the sample solution and standard solution
(1), respectively. Separately, weigh accurately about 20 mg
of Nystatin RS, add exactly 40 mL of dimethylsulfoxide to
dissolve, then add methanol to make exactly 200 mL. Pipet 4 Amphotericin B for Injection
mL of this solution, add methanol to make exactly 50 mL,
注射用アムホテリシン B
and use this solution as the standard solution (2). Perform
the test with these solutions as directed under Ultraviolet-
visible Spectrophotometry <2.24> using a solution obtained Amphotericin B for Injection is a preparation for
in the same manner as the sample solution as the blank, and injection which is dissolved before use.
determine the absorbances at 282 nm and at 304 nm. Calcu- It contains not less than 90.0z and not more than
late the amount of amphotericin A by the following equa- 120.0z of the labeled potency of amphotericin B
tion: not more than 5z for Amphotericin B used for injec- (C47H73NO17: 924.08).
tions, and not more than 15z for Amphotericin B not used
for injections.
tions, with Amphotericin B.
Amount (z) of amphotericin A
Description Amphotericin B for Injection occurs as yellow
M × {(ASa1 × AT2) － (ASa2 × AT1)} × 25
＝ S to orange, powder or masses.
MT × {(ASa1 × ASb2) － (ASa2 × ASb1)}
Identification To an amount of Amphotericin B for Injec-
MS: Amount (mg) of Nystatin RS taken
tion, equivalent to 25 mg (potency) of Amphotericin B, add
MT: Amount (mg) of Amphotericin B taken
5 mL of dimethylsulfoxide and 45 mL of methanol, and
ASa1: Absorbance at 282 nm of the standard solution (1)
shake. To 1 mL of this solution add methanol to make 50
ASb1: Absorbance at 282 nm of the standard solution (2)
mL, and filter if necessary. Determine the absorption spec-
ASa2: Absorbance at 304 nm of the standard solution (1)
trum of the solution as directed under Ultraviolet-visible
ASb2: Absorbance at 304 nm of the standard solution (2)
Spectrophotometry <2.24>: it exhibits maxima between 361
AT1: Absorbance at 282 nm of the sample solution
nm and 365 nm, between 380 nm and 384 nm and between
AT2: Absorbance at 304 nm of the sample solution
403 nm and 407 nm.
pH <2.54> Dissolve an amount of Amphotericin B for In-
um, 609C, 3 hours).
jection, equivalent to 50 mg (potency) of Amphotericin B, in
Assay Perform the test according to the Cylinder-plate 10 mL of water. To 1 mL of this solution add water to make
method as directed under Microbial Assay for Antibiotics 50 mL: 7.2 – 8.0.
<4.02> according to the following conditions.
(i) Test organism—Saccharomyces cerevisiae ATCC
of Amphotericin B for Injection, equivalent to 50 mg (po-
9763
tency) of Amphotericin B, in 10 mL of water: the solution is
(ii) Culture medium—Use the medium 2) under (1) Agar
clear and yellow to orange.
media for seed and base layer.
(iii) Preparation of cylinder-agar plate—Proceed as di- Loss on drying <2.41> Not more than 8.0z (0.3 g, in vacu-
rected in 1.5 Preparation of agar base layer plates under the um, 609C, 3 hours).
Cylinder plate method, using Petri dish plates not dispensing
the agar medium for base layer and dispensing 8.0 mL of the
tency).
seeded agar medium.
(iv) Standard solution—Use light-resistant vessels. Uniformity of dosage units <6.02> It meets the requirement
428 Amphotericin B Syrup / Official Monographs JP XVII
of the Mass variation test (T: 105.0z). make exactly 100 mL, and use this solution as the sample
stock solution. Measure exactly a suitable amount of the
sample stock solution, add dimethylsulfoxide to make solu-
tions so that each mL contains about 200 mg (potency) and
Insoluble particulate matter <6.07> It meets the require- 50 mg (potency). Pipet 1 mL each of these solutions, add 0.2
ment. mol/L phosphate buffer solution (pH 10.5) to make exactly
20 mL, and use these solutions as the high concentration
sample solution and low concentration sample solution, re-
spectively.
Assay Perform the test according to the Cylinder-plate
method as directed under Microbial Assay for Antibiotics
(i) Test organism, culture medium, preparation of cylin-
der-agar plate and standard solutions—Proceed as directed
in the Assay under Amphotericin B. Amphotericin B Tablets
(ii) Sample solutions—Prepare using light-resistant con-
アムホテリシン B 錠
tainers. Weigh accurately an amount of Amphotericin B for
Injection, equivalent to about 50 mg (potency), dissolve in
dimethylsulfoxide to make exactly 50 mL, and use this solu- Amphotericin B Tablets contain not less than 90.0z
tion as the sample stock solution. Measure exactly a suitable and not more than 120.0z of the labeled potency of
quantity of the sample stock solution, add dimethylsulfoxide amphotericin B (C47H73NO17: 924.08).
to make solutions so that each mL contains about 200 mg
(potency) and 50 mg (potency). Pipet 1 mL each of these so-
with Amphotericin B.
lutions, add 0.2 mol/L phosphate buffer solution (pH 10.5)
to make exactly 20 mL, and use these solutions as the high Identification To an amount of pulverized Amphotericin B
concentration sample solution and low concentration sample Tablets, equivalent to 25 mg (potency) of Amphotericin B,
solution, respectively. add 5 mL of dimethylsulfoxide and 45 mL of methanol, and
shake. To 1 mL of this solution add methanol to make 50
mL, and filter, if necessary. Determine the absorption spec-
Storage—Light-resistant, at a cold place.
trum of the solution as directed under Ultraviolet-visible
nm and 365 nm, between 380 nm and 384 nm and between
Amphotericin B Syrup 403 nm and 407 nm.
アムホテリシン B シロップ Loss on drying <2.41> Not more than 5.0z (0.3 g, in vacu-
um, 609C, 3 hours).
Amphotericin B Syrup contain not less than 90.0z Uniformity of dosage units <6.02> It meets the requirement
and not more than 115.0z of the labeled potency of of the Mass variation test (T: 105.0z).
amphotericin B (C47H73NO17: 924.08).
Method of preparation Prepare as directed under Syrup, method as directed under Microbial Assay for Antibiotics
with Amphotericin B. <4.02> according to the following conditions.
Identification To an amount of Amphotericin B Syrup,
equivalent to 25 mg (potency) of Amphotericin B, add 5 mL
in the Assay under Amphotericin B.
of dimethylsulfoxide and 45 mL of methanol, and shake. To
1 mL of this solution add methanol to make 50 mL, and
tainers. Weigh accurately and powder not less than 20 tablets
filter, if necessary. Determine the absorption spectrum of the
of Amphotericin B Tablets. Weigh accurately a part of the
powder, equivalent to about 0.1 g (potency), add about 70
tometry <2.24>: it exhibits maxima between 361 nm and 365
mL of dimethylsulfoxide, shake, then add dimethylsulfoxide
nm, between 380 nm and 384 nm and between 403 nm and
to make exactly 100 mL, centrifuge, and use the supernatant
407 nm.
liquid as the sample stock solution. Measure exactly a suita-
pH <2.54> 5.0 – 7.0 ble amount of the sample stock solution, add dimethylsul-
foxide to make solutions so that each mL contains 200 mg
Microbial limit <4.05> The acceptance criteria of TAMC
(potency) and 50 mg (potency). Pipet 1 mL each of these so-
and TYMC are 102 CFU/mL and 5 × 101 CFU/mL, respec-
lutions, add 0.2 mol/L phosphate buffer solution (pH 10.5)
tively.
to make exactly 20 mL, and use these solutions as the high
Assay Perform the test according to the Cylinder-plate concentration sample solution and low concentration sample
method as directed under Microbial Assay for Antibiotics solution, respectively.
ers.
in the Assay under Amphotericin B.
tainers. Weigh accurately an amount of Amphotericin B
Syrup, equivalent to about 0.1 g (potency), add about 70 mL
of dimethylsulfoxide, shake, then add dimethylsulfoxide to
JP XVII Official Monographs / Anhydrous Ampicillin 429
Anhydrous Ampicillin System performance, and system repeatability: Proceed as
Anhydrous Aminobenzylpenicillin Test for required detectability: To exactly 1 mL of the
standard solution add the mobile phase to make exactly 10
無水アンピシリン mL. Confirm that the peak area of ampicillin obtained from
tained from 10 mL of the standard solution.
Assay Weigh accurately an amount of Anhydrous Ampicil-
lin and Ampicillin RS, equivalent to about 50 mg (potency),
C16H19N3O4S: 349.40
add exactly 5 mL each of the internal standard solution and
(2S,5R,6R)-6-[(2R)-2-Amino-2-phenylacetylamino]-
the mobile phase to make 50 mL, and use these solutions as
3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-
the sample solution and standard solution. Perform the test
carboxylic acid
[69-53-4]
to the following conditions, and calculate the ratios, QT and
Anhydrous Ampicillin contains not less than 960 mg
QS, of the peak area of ampicillin to that of the internal
standard.
Anhydrous Ampicillin is expressed as mass (potency) Amount [ mg (potency)] of ampicillin (C16H19N3O4S)
of ampicillin (C16H19N3O4S). ＝ MS × QT/QS × 1000
Description Anhydrous Ampicillin occurs as white to light MS: Amount [mg (potency)] of Ampicillin RS taken
yellowish white, crystals or crystalline powder.
Internal standard solution—A solution of guaifenesin in the
It is sparingly soluble in water, slightly soluble in metha-
mobile phase (1 in 200).
nol, very slightly soluble in ethanol (95), and practically in-
soluble in acetonitrile.
Identification Determine the infrared absorption spectrum length: 230 nm).
of Anhydrous Ampicillin as directed in the potassium bro- Column: A stainless steel column 4.6 mm in inside diame-
mide disk method under Infrared Spectrophotometry <2.25>, ter and 15 cm in length, packed with octadecylsilanized silica
and compare the spectrum with the Reference Spectrum: gel for liquid chromatography (5 mm in particle diameter).
both spectra exhibit similar intensities of absorption at the Column temperature: A constant temperature of about
same wave numbers. 259C.
Mobile phase: Dissolve 5.94 g of diammonium hydrogen
D : ＋280 – ＋3059(0.5 g calcu-
phosphate in 850 mL of water, add 100 mL of acetonitrile,
adjust the pH to 5.0 with phosphoric acid, and add water to
pH <2.54> The pH of a solution obtained by dissolving make exactly 1000 mL.
1.0 g of Anhydrous Ampicillin in 100 mL of water is be- Flow rate: Adjust so that the retention time of ampicillin
tween 4.0 and 5.5. is about 6 minutes.
Anhydrous Ampicillin according to Method 2, and perform
ditions, ampicillin and the internal standard are eluted in this
than 40.
of Anhydrous Ampicillin according to Method 3, and per-
form the test (not more than 2 ppm).
(3) Related substances—Dissolve 0.05 g of Anhydrous
Ampicillin in the mobile phase to make 50 mL, and use this
of the peak area of ampicillin to that of the internal standard
solution as the sample solution. Pipet 1 mL of the sample so-
lution, add the mobile phase to make exactly 100 mL, and
use this solution as the standard solution. Perform the test Containers and storage Containers—Tight containers.
peak area by the automatic integration method: the area of
each peak other than ampicillin from the sample solution is
not larger than the peak area of ampicillin from the standard
solution.
the Assay.
the retention time of ampicillin.
430 Ampicillin Hydrate / Official Monographs JP XVII
Ampicillin Hydrate System performance, and system repeatability: Proceed as
Aminobenzylpenicillin Test for required detectability: Measure exactly 1 mL of
the standard solution, and add the mobile phase to make
アンピシリン水和物 exactly 10 mL. Confirm that the peak area of ampicillin
obtained from 10 mL of this solution is equivalent to 7 to
13z of that obtained from 10 mL of the standard solution.
(4) N, N-Dimethylaniline—Weigh accurately about 1 g of
Ampicillin Hydrate, dissolve in 5 mL of sodium hydroxide
TS, add exactly 1 mL of the internal standard solution,
shake vigorously for 1 minute, and use the upper layer liquid
obtained after allowing it to stand as the sample solution.
C16H19N3O4S.3H2O: 403.45
Separately, weigh accurately about 50 mg of N, N-
(2S,5R,6R)-6-[(2R)-2-Amino-2-phenylacetylamino]-
dimethylaniline, dissolve in 2 mL of hydrochloric acid and
3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-
20 mL of water, add water to make exactly 50 mL, and use
carboxylic acid trihydrate
this solution as the standard stock solution. Pipet 5 mL of
[7177-48-2]
the standard stock solution, and add water to make exactly
250 mL. Pipet 1 mL of this solution, add 5 mL of sodium
Ampicillin Hydrate contains not less than 960 mg
hydroxide TS and exactly 1 mL of the internal standard solu-
tion, shake vigorously for 1 minute, and use the upper layer
liquid obtained after allowing it to stand as the standard
Ampicillin Hydrate is expressed as mass (potency) of
solution. Perform the test with 1 mL each of the sample
ampicillin (C16H19N3O4S: 349.40).
solution and standard solution as directed under Gas Chro-
Description Ampicillin Hydrate occurs as a white to light matography <2.02> according to the following conditions,
yellowish white, crystals or crystalline powder. calculate the ratios, QT and QS, of the peak area of N, N-
It is sparingly soluble in water, slightly soluble in metha- dimethylaniline to that of the internal standard, and calcu-
nol, very slightly soluble in ethanol (95), and practically in- late the amount of N, N-dimethylaniline by the following
soluble in acetonitrile. equation: not more than 20 ppm.
Identification Determine the infrared absorption spectrum Amount (ppm) of N, N-dimethylaniline
of Ampicillin Hydrate as directed in the potassium bromide ＝ MS/MT × QT/QS × 400
disk method under Infrared Spectrophotometry <2.25>, and
MS: Amount (g) of N, N-dimethylaniline taken
MT: Amount (g) of Ampicillin Hydrate taken
spectrum of Ampicillin RS: both spectra exhibit similar in-
tensities of absorption at the same wave numbers. Internal standard solution—A solution of naphthalene in
cyclohexane (1 in 20,000).
＋280 – ＋3059(0.5 g calcu-
D:
pH <2.54> The pH of a solution obtained by dissolving Column: A glass column 2.6 mm in inside diameter and
1.0 g of Ampicillin Hydrate in 400 mL of water is between 2 m in length, packed with siliceous earth for gas chromatog-
3.5 and 5.5. raphy (180 – 250 mm in particle diameter) coated with 50z
phenyl-50z methyl polysiloxane for gas chromatography at
the ratio of 3z.
Ampicillin Hydrate according to Method 2, and perform the
1209C.
Flow rate: Adjust so that the retention time of N, N-
of Ampicillin hydrate according to Method 3, and perform
dimethylaniline is about 5 minutes.
the test (not more than 2 ppm).
(3) Related substances—Dissolve 50 mg of Ampicillin
hydrate in the mobile phase to make 50 mL, and use this so-
the standard stock solution, and add water to make exactly
lution as the sample solution. Pipet 1 mL of the sample solu-
250 mL. Pipet 1 mL of this solution, add 5 mL of sodium
tion, add the mobile phase to make exactly 100 mL, and use
hydroxide TS and exactly 1 mL of the internal standard solu-
tion, shake vigorously for 1 minute, and use the upper layer
exactly 10 mL each of the sample solution and standard solu-
liquid obtained after allowing it to stand for the test. Con-
tion as directed under Liquid Chromatography <2.01> ac-
firm that when the procedure is run with 1 mL of the upper
cording to the following conditions, and determine each
layer liquid under the above operating conditions, the ratio
of the peak area of N, N-dimethylaniline to that of the inter-
the peak other than ampicillin obtained from the sample so-
nal standard is equivalent to 15 to 25z of the ratio of the
lution is not larger than the peak area of ampicillin obtained
peak area of N, N-dimethylaniline to that of the internal
standard obtained from the standard solution.
System performance: Dissolve 50 mg of N, N-dimethylani-
line in cyclohexane to make 50 mL. To 1 mL of this solution
add the internal standard solution to make 50 mL, and use
the Assay.
this solution as the solution for system suitability test. When
the procedure is run with 1 mL of the solution for system
retention time of ampicillin.
JP XVII Official Monographs / Ampicillin Sodium 431
suitability test under the above operating conditions, N, N-

dimethylaniline and the internal standard are eluted in this Ampicillin Sodium
than 3. Aminobenzylpenicillin Sodium
with 1 mL of the solution for system suitability test under the アンピシリンナトリウム
above operating conditions, the relative standard deviation
of the ratios of the peak area of N, N-dimethylaniline to that
of the internal standard is not more than 2.0z.
Water <2.48> 12.0 – 15.0z (0.1 g, volumetric titration,
direct titration).
Assay Weigh accurately an amount of Ampicillin Hydrate
C16H18N3NaO4S: 371.39
and Ampicillin RS, equivalent to about 50 mg (potency), dis-
Monosodium (2S,5R,6R)-6-[(2R)-2-amino-2-
solve in a suitable volume of the mobile phase, add exactly 5
phenylacetylamino]-3,3-dimethyl-7-oxo-4-thia-1-
mL each of the internal standard solution and the mobile
azabicyclo[3.2.0]heptane-2-carboxylate
phase to make 50 mL, and use these solutions as the sample
[69-52-3]
solution and standard solution. Perform the test with 10 mL
Ampicillin Sodium contains not less than 850 mg
(potency) and not more than 950 mg (potency) per mg,
calculated on the anhydrous basis. The potency of
the peak area of ampicillin to that of the internal standard.
Ampicillin Sodium is expressed as mass (potency) of
Amount [ mg (potency)] of ampicillin (C16H19N3O4S) ampicillin (C16H19N3O4S: 349.40).
＝ MS × QT/QS × 1000
Description Ampicillin Sodium occurs as white to light
MS: Amount [mg (potency)] of Ampicillin RS taken yellowish white, crystals or crystalline powder.
It is very soluble in water, and sparingly soluble in ethanol
(99.5).
Operating conditions— Identification (1) Determine the infrared absorption spec-
Detector: An ultraviolet absorption photometer (wave- trum of Ampicillin Sodium, previously dried in a desiccator
length: 230 nm). (reduced pressure not exceeding 0.67 kPa, 609 C) for 3 hours,
Column: A stainless steel column 4.6 mm in inside diame- as directed in the potassium bromide disk method under
ter and 15 cm in length, packed with octadecylsilanized silica Infrared Spectrophotometry <2.25>, and compare the spec-
gel for liquid chromatography (5 mm in particle diameter). trum with the Reference Spectrum: both spectra exhibit simi-
Column temperature: A constant temperature of about lar intensities of absorption at the same wave numbers.
259 C. (2) Ampicillin Sodium responds to the Qualitative Tests
Mobile phase: Dissolve 5.94 g of diammonium hydrogen- <1.09> (1) for sodium salt.
phosphate in 850 mL of water, add 100 mL of acetonitrile,
D : ＋246 – ＋2729(1 g calculated
adjust the pH to 5.0 with phosphoric acid, and add water to
on the anhydrous basis, water, 100 mL, 100 mm).
make exactly 1000 mL.
Flow rate: Adjust so that the retention time of ampicillin pH <2.54> The pH of a solution obtained by dissolving
is about 6 minutes. 1.0 g of Ampicillin Sodium in 10 mL of water is between 8.0
System suitability— and 10.0.
(potency) of Ampicillin Sodium in 0.75 mL of water: the so-
ditions, ampicillin and the internal standard are eluted in this
lution is clear, and its absorbance at 400 nm, determined as
than 40.
is not more than 0.40.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Ampicil-
lin Sodium according to Method 1, and perform the test.
of the peak area of ampicillin to that of the internal standard
Containers and storage Containers—Tight containers. of Ampicillin Sodium according to Method 1, and perform
(4) Related substances—Dissolve 50 mg of Ampicillin
Sodium in 50 mL of the mobile phase, and use this solution
as the sample solution. Pipet 1 mL of the sample solution,
add the mobile phase to make exactly 100 mL, and use this
solution as the standard solution. Perform the test with
the peaks other than ampicillin obtained from the sample so-
lution is not larger than the peak area of ampicillin obtained
432 Ampicillin Sodium for Injection / Official Monographs JP XVII
from the standard solution. 35.
Operating conditions— System repeatability: When the test is repeated 6 times
Detector, column, column temperature, mobile phase, and with 10 mL of the standard solution under the above operat-
flow rate: Proceed as directed in the operating conditions in ing conditions, the relative standard deviation of the ratio of
the Assay. the peak area of ampicillin to that of the internal standard is
Time span of measurement: About 10 times as long as the not more than 1.0z.
retention time of ampicillin.
exactly 10 mL. Confirm that the peak area of ampicillin Ampicillin Sodium for Injection
注射用アンピシリンナトリウム
13z of that of ampicillin obtained from 10 mL of the stand-
ard solution.
System performance: Dissolve 50 mg of Ampicillin RS in a Ampicillin Sodium for Injection is a preparation for
suitable amount of the mobile phase, add 5 mL of a solution injection which is dissolved before use.
of guaifenesin in the mobile phase (1 in 200) and the mobile It contains not less than 90.0z and not more
phase to make 50 mL, and use this solution as the solution than 110.0z of the labeled potency of ampicillin
for system suitability test. When the procedure is run with 10 (C16H19N3O4S: 349.40).
operating conditions, ampicillin and guaifenesin are eluted
tions, with Ampicillin Sodium.
in this order with the resolution between these peaks being
not less than 35. Description Ampicillin Sodium for Injection occurs as
System repeatability: When the test is repeated 6 times white to light yellowish white, crystals or crystalline powder.
Identification Proceed as directed in the Identification (1)
under Ampicillin Sodium.
tion of the peak area of ampicillin is not more than 1.0z.
pH <2.54> The pH of a solution prepared by dissolving an
Assay Weigh accurately an amount of Ampicillin Sodium
amount of Ampicillin Sodium for Injection, equivalent to
and Ampicillin RS, equivalent to about 50 mg (potency),
1.0 g (potency) of Ampicillin Sodium, in 10 mL of water is
dissolve them in a suitable amount of the mobile phase, add
8.0 to 10.0.
exactly 5 mL of the internal standard solution, then add the
mobile phase to make 50 mL, and use these solutions as the Purity Clarity and color of solution—Dissolve an amount
sample solution and the standard solution, respectively. Per- of Ampicillin Sodium for Injection, equivalent to 0.25 g
form the test with 10 mL each of the sample solution and (potency) of Ampicillin Sodium, in 0.75 mL of water: the
standard solution as directed under Liquid Chromatography solution is clear. Perform the test with the solution as di-
<2.01> according to the following conditions, and calculate rected under Ultraviolet-visible Spectrophotometry <2.24>:
the ratios, QT and QS, of the peak area of ampicillin to that the absorbance at 400 nm is not more than 0.40.
of the internal standard.
Amount [mg (potency)] of ampicillin (C16H19N3O4S) tion, direct titration).
＝ MS × QT/QS × 1000
MS: Amount [mg (potency)] of Ampicillin RS taken tency).
Internal standard solution—A solution of guaifenesin in the Uniformity of dosage units <6.02> It meets the requirement
mobile phase (1 in 200). of the Mass variation test.
length: 230 nm).
Column: A stainless steel column 4.6 mm in inside diame- Insoluble particulate matter <6.07> It meets the require-
ter and 15 cm in length, packed with octadecylsilanized silica ment.
259 C.
Mobile phase: Dissolve 5.94 g of diammonium hydrogen Assay Weigh accurately the mass of the contents of not less
phosphate in 850 mL of water, add 100 mL of acetonitrile, than 10 containers of Ampicillin Sodium for Injection.
adjust to pH 5.0 with phosphoric acid, and add water to Weigh accurately an amount of a portion of the contents,
make 1000 mL. equivalent to about 50 mg (potency) of Ampicillin Sodium,
Flow rate: Adjust so that the retention time of ampicillin add exactly 5 mL of the internal standard solution and dis-
is about 6 minutes. solve. Then add the mobile phase to make 50 mL, and use
System suitability— this solution as the sample solution. Separately, weigh accu-
System performance: When the procedure is run with 10 rately an amount of Ampicillin RS, equivalent to about 50
mL of the standard solution under the above operating con- mg (potency), add exactly 5 mL of the internal standard so-
ditions, ampicillin and guaifenesin are eluted in this order lution and dissolve. Then add the mobile phase to make 50
with the resolution between these peaks being not less than mL, and use this solution as the standard solution. Perform
JP XVII Official Monographs / Ampicillin Sodium and Sulbactam Sodium for Injection 433
the test with 10 mL each of the sample solution and standard Operating conditions—
solution as directed under Liquid Chromatography <2.01> Column, column temperature, mobile phase, and flow
according to the following conditions, and calculate the rate: Proceed as directed in the operating conditions in the
ratios, QT and QS, of the peak area of ampicillin to that of Assay.
the internal standard. Detector: An ultraviolet absorption photometer (wave-
length: 230 nm).
Amount [mg (potency)] of ampicillin (C16H19N3O4S)
＝ M S × Q T / QS
MS: Amount [mg (potency)] of Ampicillin RS taken suitability in the Assay.
(2) The retention times of sulbactam obtained from the
sample solution and the standard solution observed in the
Assay are the same, and the peak area of sulbactam observed
in the Assay obtained from the sample solution is 2.0 to 2.6
times the peak area of sulbactam observed in the test per-
length: 230 nm).
formed with 10 mL of the sample solution obtained in the
Assay as directed under Liquid Chromatography <2.01> ac-
cording to the following conditions.
Column, column temperature, mobile phase, and flow
259 C.
rate: Proceed as directed in the operating conditions in the
Mobile phase: Dissolve 5.94 mg of diammonium hydrogen
Assay.
phosphate in 850 mL of water, add 100 mL of acetonitril,
add phosphoric acid to adjust the pH to 5.0, then add water
length: 230 nm).
to make exactly 1000 mL.
Flow rate: Adjust so that the retention time of ampicillin
is about 6 minutes.
System performance: When the procedure is run with 10 pH <2.54> The pH of a solution prepared by dissolving an
mL of the standard solution under the above operating con- amount of Ampicillin Sodium and Sulbactam Sodium for
ditions, ampicillin and the internal standard are eluted in this Injection, equivalent to 1.0 g (potency) of ampicillin
order with the resolution between these peaks being not less (C16H19N3O4S), in 10 mL of water is between 8.0 and 10.0.
than 26.
Purity (1) Clarity and color of solution—Dissolve an
amount of Ampicillin Sodium and Sulbactam Sodium for
Injection, equivalent to 1.0 g (potency) of ampicillin
(C16H19N3O4S), in 10 mL of water: the solution is clear.
the peak area of ampicillin to that of the internal standard is
Determine the absorption of this solution as directed under
not more than1.0z.
Ultraviolet-visible Spectrophotometry <2.24>: the absor-
Containers and storage Containers—Hermetic containers. bance at 425 nm is not more than 0.10.
(2) Total penicilloic acid—Weigh accurately about 25 mg
of Ampicillin Sodium and Sulbactam Sodium for Injection,
Ampicillin Sodium and Sulbactam place in a glass-stoppered flask, dissolve in 25 mL of 0.02
mol/L phosphate buffer solution (pH 3.0), add exactly 5 mL
Sodium for Injection of 0.005 mol/L iodine VS, stopper the flask, allow to stand
for 5 minutes, and titrate <2.50> with 0.005 mol/L sodium
注射用アンピシリンナトリウム･スルバクタムナトリウム
thiosulfate VS (indicator: 1.0 mL of starch TS). Perform a
blank determination in the same manner, and make any
Ampicillin Sodium and Sulbactam Sodium for In- necessary correction: the amount of total penicilloic acid
jection is a preparation for injection which is dissolved (as C16H21N3O5S: 367.42) is not more than 3.0z.
before use.
Each mL of 0.005 mol/L sodium thiosulfate VS
It contains not less than 95.0z and not more ＝ 0.2064 mg of C16H21N3O5S
than 112.0z of the labeled potency of ampicillin
(C16H19N3O4S: 349.40) and sulbactam (C8H11NO5S: Water <2.48> Not more than 2.0z (0.5 g, volumetric titra-
233.24). tion, direct titration).
Method of preparation Prepare as directed under Injec- Bacterial endotoxins <4.01> Less than 0.10 EU/mg (po-
tions, with Ampicillin Sodium and Sulbactam Sodium. tency).
Description Ampicillin Sodium and Sulbactam Sodium for Uniformity of dosage units <6.02> Perform the test accord-
Injection occurs as a white to yellowish white powder. ing to the following method: it meets the requirement of the
Content uniformity test (T: 105.0z).
Identification (1) The retention times of ampicillin ob-
Dissolve 1 Ampicillin Sodium and Sulbactam Sodium for
tained from the sample solution and the standard solution
Injection in the mobile phase to make exactly V mL so
observed in the Assay are the same, and the peak area of am-
that each mL contains 5 mg (potency) of ampicillin
picillin observed in the Assay obtained from the sample solu-
(C16H19N3O4S). Pipet 5 mL of this solution, add exactly 5
tion is 2.8 to 3.6 times the peak area of ampicillin observed
mL of the internal standard solution, then add the mobile
in the test performed with 10 mL of the sample solution ob-
phase to make 50 mL, and use this solution as the sample so-
tained in the Assay as directed under Liquid Chromatogra-
lution. Then, proceed as directed in the Assay.
phy <2.01> according to the following conditions.
434 Ampiroxicam / Official Monographs JP XVII
Amount [mg (potency)] of ampicillin (C16H19N3O4S) peaks is not less than 2.0.
＝ MS1 × QTa/QSa × V/10 System repeatability: When the test is repeated 6 times
Amount [mg (potency)] of sulbactam (C8H11NO5S)
＝ MS2 × QTb/QSb × V/10
area of sulbactam is not more than 1.0z.
MS1: Amount [mg (potency)] of Ampicillin RS taken
Containers and storage—Hermetic containers. Plastic con-
MS2: Amount [mg (potency)] of Sulbactam RS taken
tainers for aqueous injections may be used.
Foreign insoluble matter <6.06> Perform the test according Ampiroxicam
アンピロキシカム
ment.
Assay Weigh accurately the mass of the contents of not less
than 10 containers of Ampicillin Sodium and Sulbactam
Sodium for Injection. Weigh accurately an amount of a por-
tion of the contents, equivalent to about 0.25 g (potency) of C20H21N3O7S: 447.46
ampicillin (C16H19N3O4S), and dissolve in the mobile phase Ethyl (1RS )-1-({2-methyl-1,1-dioxido-3-[(pyridin-2-
to make exactly 50 mL. Pipet 5 mL of this solution, add ex- ylamino)carbonyl]-2H-1,2-benzothiazin-4-yl}oxy)ethyl carbonate
actly 5 mL of the internal standard solution, add the mobile [99464-64-9]
phase to make 50 mL, and use this solution as the sample so-
lution. Separately, weigh accurately an amount of Ampicil- Ampiroxicam, when dried, contains not less than
lin RS, equivalent to about 50 mg (potency), and an amount 99.0z and not more than 101.0z of ampiroxicam
of Sulbactam RS, equivalent to about 25 mg (potency), dis- (C20H21N3O7S).
solve in the mobile phase, add exactly 10 mL of the internal
Description Ampiroxicam occurs as a white to yellowish
standard solution, add the mobile phase to make 100 mL,
It is freely soluble in acetic acid (100), soluble in aceto-
nitrile, very slightly soluble in ethanol (99.5), and practically
insoluble in water.
A solution of Ampiroxicam in acetonitrile (1 in 20) shows
QTa and QTb, of the peak areas of ampicillin and sulbactam
to that of the internal standard obtained from the sample
It shows crystal polymorphism.
solution, and the ratios, QSa and QSb, of the peak areas of
ampicillin and sulbactam to that of the internal standard Identification (1) Determine the absorption spectrum of a
obtained from the standard solution. solution of Ampiroxicam in 0.01 mol/L hydrochloric acid-
methanol TS (1 in 100,000) as directed under Ultraviolet-
Amount [mg (potency)] of ampicillin (C16H19N3O4S)
＝ MS1 × QTa/QSa × 5
Amount [mg (potency)] of sulbactam (C8H11NO5S) tensities of absorption at the same wavelengths.
＝ MS2 × QTb/QSb × 5 (2) Determine the infrared absorption spectrum of Am-
piroxicam as directed in the potassium bromide disk method
MS1: Amount [mg (potency)] of Ampicillin RS taken
MS2: Amount [mg (potency)] of Sulbactam RS taken
Internal standard solution—A solution of parahydroxyben- similar intensities of absorption at the same wave numbers.
Ampiroxicam according to Method 2, and perform the test.
length: 215 mn).
solution (not more than 20 ppm).
(2) Related substances—Dissolve 20 mg of Ampiroxicam
in 50 mL of acetonitrile, and use this solution as the sample
solution. Pipet 1 mL of the sample solution, add acetonitrile
to make exactly 200 mL, and use this solution as the stand-
359 C.
ard solution. Perform the test with exactly 10 mL each of the
Mobile phase: A mixture of 0.02 mol/L phosphate buffer
(pH 3.0) and acetonitrile for liquid chromatography (23:2).
Flow rate: Adjust so that the retention time of the internal
standard is about 9 minutes.
tegration method: the area of the peak, having a relative
retention time of about 0.17 to ampiroxicam, obtained from
the sample solution is not larger than 1/2 times the peak area
of ampiroxicam obtained from the standard solution, the
ditions, sulbactam, the internal standard and ampicillin are
area of the peak other than ampiroxicam and the peak men-
eluted in this order, and either resolution between these
tioned above from the sample solution is not larger than 2/5
JP XVII Official Monographs / Ampiroxicam Capsules 435
times the peak area of ampiroxicam from the standard solu- methanol TS, shake well, and centrifuge. To 5 mL of the su-
tion, and the total area of the peaks other than ampiroxicam pernatant liquid add 0.01 mol/L hydrochloric acid-methanol
from the sample solution is not larger than the peak area of TS to make 50 mL. Determine the absorption spectrum of
ampiroxicam from the standard solution. For the area of the this solution as directed under Ultraviolet-visible Spectro-
peaks, having the relative retention time of about 0.17 and photometry <2.24>: it exhibits a maximum between 318 nm
about 0.46 to ampiroxicam, multiply the relative response and 322 nm.
factor, 0.37 and 0.60, respectively.
length: 254 nm).
Take out the contents of 1 capsule of Ampiroxicam Cap-
sules, add acetonitrile to make exactly V mL so that each mL
contains about 0.27 mg of ampiroxicam (C20H21N3O7S). Stir
for 30 minutes, then centrifuge, and use the supernatant liq-
uid as the sample solution. Then, proceed as directed in the
259 C.
Assay.
500), methanol and acetonitrile (5:3:2). Amount (mg) of ampiroxicam (C20H21N3O7S)
Flow rate: Adjust so that the retention time of ampiroxic- ＝ MS × AT/AS × V/100
am is about 9 minutes.
MS: Amount (mg) of ampiroxicam for assay taken
retention time of ampiroxicam, beginning after the solvent Dissolution <6.10> When the test is performed at 50 revolu-
peak. tions per minute according to the Paddle method using the
System suitability— sinker, using 900 mL of 1st fluid for dissolution test as the
Test for required detectability: Pipet 5 mL of the standard dissolution medium, the dissolution rate in 30 minutes of
solution, add acetonitrile to make exactly 50 mL. Confirm Ampiroxicam Capsules is not less than 70z.
that the peak area of ampiroxicam obtained with 10 mL of Start the test with 1 capsule of Ampiroxicam Capsules,
this solution is equivalent to 7 to 13z of that obtained with withdraw not less than 20 mL of the medium at the specified
10 mL of the standard solution. minute after starting the test, and filter through a membrane
System performance: When the procedure is run with 10 filter with a pore size not exceeding 0.45 mm. Discard the
mL of the standard solution under the above operating con- first 10 mL of the filtrate, pipet V mL of the subsequent fil-
ditions, the number of theoretical plates and the symmetry trate, add the dissolution medium to make exactly V? mL so
factor of the peak of ampiroxicam are not less than 3000 and that each mL contains about 15 mg of ampiroxicam
not more than 2.0, respectively. (C20H21N3O7S), and use this solution as the sample solution.
System repeatability: When the test is repeated 6 times Separately, weigh accurately about 30 mg of ampiroxicam
with 10 mL of the standard solution under the above operat- for assay, previously dried at 1059C for 3 hours, dissolve in
ing conditions, the relative standard deviation of the peak 5 mL of acetonitrile, and add the dissolution medium to
area of ampiroxicam is not more than 5z. make exactly 100 mL. Pipet 5 mL of this solution, add the
dissolution medium to make exactly 100 mL, and use this so-
lution as the standard solution. Determine the absorbances,
3 hours).
Residue on ignition <2.44> Not more than 0.2z (1 g). solution as directed under Ultraviolet-visible Spectropho-
tometry <2.24>, using the dissolution medium as the blank.
Assay Weigh accurately about 0.22 g of Ampiroxicam,
previously dried, dissolve in 50 mL of acetic acid (100), and Dissolution rate (z) with respect to the labeled amount of
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- ampiroxicam (C20H21N3O7S)
metric titration). Perform a blank determination in the same ＝ MS × AT/AS × V?/V × 1/C × 45
MS: Amount (mg) of ampiroxicam for assay taken
Each mL of 0.1 mol/L perchloric acid VS C: Labeled amount (mg) of ampiroxicam (C20H21N3O7S)
＝ 44.75 mg of C20H21N3O7S in 1 capsule
Containers and storage Containers—Tight containers. Assay Take out the contents of not less than 20 Ampiroxic-
Storage—Light-resistant. am Capsules, weigh accurately the mass of the contents, and
powder if necessary. Weigh accurately a portion of the
powder, equivalent to about 13.5 mg of ampiroxicam
Ampiroxicam Capsules (C20H21N3O7S), and add acetonitrile to make exactly 50 mL.
Stir for 30 minutes, centrifuge, and use the supernatant
アンピロキシカムカプセル liquid as the sample solution. Separately, weigh accurately
about 27 mg of ampiroxicam for assay, previously dried at
1059C for 3 hours, dissolve in acetonitrile to make exactly
Ampiroxicam Capsules contain not less than 95.0z
ampiroxicam (C20H21N3O7S: 447.46).
Method of Preparation Prepare as directed under Cap- raphy <2.01> according to the following condition, and de-
sules, with Ampiroxicam. termine the peak areas, AT and AS, of ampiroxicam in each
solution.
Identification Take out the contents of Ampiroxicam Cap-
sules, to a quantity of the contents, equivalent to 10 mg of Amount (mg) of ampiroxicam (C20H21N3O7S)
Ampiroxicam, add 100 mL of 0.01 mol/L hydrochloric acid- ＝ MS × AT/AS × 1/2
436 Amyl Nitrite / Official Monographs JP XVII
MS: Amount (mg) of ampiroxicam for assay taken Add 1.0 mL of Amyl Nitrite, and warm between 609C and
709C for 1 minute: a brown to black color is not produced.
(4) Residue on evaporation—Evaporate 10.0 mL of
Amyl Nitrite on a water bath in a draft chamber, carefully
length: 254 nm).
protecting from flame, and dry the residue at 1059C for 1
hour: the mass of the residue is not more than 1.0 mg.
gel for liquid chromatography (4 mm in particle diameter). Assay Weigh accurately a volumetric flask containing 10
Column temperature: A constant temperature of about mL of ethanol (95), add about 0.5 g of Amyl Nitrite, and
259 C. weigh accurately again. Add exactly 25 mL of 0.1 mol/L
Mobile phase: A mixture of diluted acetic acid (100) (3 in silver nitrate VS, then add 15 mL of potassium chlorate solu-
500), methanol, and acetonitrile (5:3:2). tion (1 in 20) and 10 mL of dilute nitric acid, stopper the
Flow rate: Adjust so that the retention time of ampiroxic- flask immediately, and shake it vigorously for 5 minutes.
am is about 9 minutes. Dilute with water to make exactly 100 mL, shake, and filter
System suitability— through dry filter paper. Discard the first 20 mL of the fil-
System performance: When the procedure is run with 10 trate, measure exactly 50 mL of the subsequent filtrate, and
mL of the standard solution under the above operating con- titrate <2.50> the excess silver nitrate with 0.1 mol/L ammo-
ditions, the number of theoretical plates and the symmetry nium thiocyanate VS (indicator: 2 mL of ammonium iron
factor of the peak of ampiroxicam are not less than 4000 and (III) sulfate TS). Perform a blank determination.
Each mL of 0.1 mol/L silver nitrate VS
＝ 35.15 mg of C5H11NO2
ing conditions, the relative standard deviation of the peak Containers and storage Containers—Hermetic containers
area of ampiroxicam is not more than 1.0z. not exceeding 10–ml capacity.
Storage—Light-resistant, in a cold place, and remote from
fire.
ers.
Amyl Nitrite Dental Antiformin

亜硝酸アミル
Dental Sodium Hypochlorite Solution
歯科用アンチホルミン
C5H11NO2: 117.15
Dental Antiformin contains not less than 3.0 w/vz
Amyl Nitrite is the nitrous acid ester of 3-methyl-
and not more than 6.0 w/vz of sodium hypochlorite
butanol-1 and contains a small quantity of 2-methyl-
(NaClO: 74.44).
butanol-1 and the nitrous acid esters of other homo-
logues. Description Dental Antiformin is a slightly light yellow-
It contains not less than 90.0z of amyl nitrite green, clear liquid. It has a slight odor of chlorine.
(C5H11NO2). It gradually changes by light.
Description Amyl Nitrite is a clear, light yellowish liquid, Identification (1) Dental Antiformin changes red litmus
and has a characteristic, fruity odor. paper to blue, and then decolorizes it.
It is miscible with ethanol (95), and with diethyl ether. (2) To Dental Antiformin add dilute hydrochloric acid:
It is practically insoluble in water. it evolves the odor of chlorine, and the gas changes potas-
It is affected by light and by heat. sium iodide starch paper moistened with water to blue.
It is volatile at ordinary temperature and flammable even (3) Dental Antiformin responds to the Qualitative Tests
at a low temperature. <1.09> (1) for sodium salt.
Boiling point: about 979C
Assay Measure exactly 3 mL of Dental Antiformin in a
Identification Determine the infrared spectrum of Amyl glass-stoppered flask, add 50 mL of water, 2 g of potassium
Nitrite as directed in the liquid film method under Infrared iodide and 10 mL of acetic acid (31), and titrate <2.50> the
Spectrophotometry <2.25>, and compare the spectrum with liberated iodine with 0.1 mol/L sodium thiosulfate VS
the Reference Spectrum: both spectra exhibit similar intensi- (indicator: 3 mL of starch TS).
ties of absorption at the same wave numbers.
Each mL of 0.1 mol/L sodium thiosulfate VS
Specific gravity <2.56> d 20
20: 0.871 – 0.880 ＝ 3.722 mg of NaClO
Purity (1) Acidity—To 5 mL of Amyl Nitrite add a mix- Containers and storage Containers—Tight containers.
ture of 1.0 mL of 1 mol/L sodium hydroxide VS, 10 mL of Storage—Light-resistant, and not exceeding 109C.
water and 1 drop of phenolphthalein TS, shake, and allow to
stand for 1 minute: the light red color of the water layer does
not disappear.
(2) Water—Allow 2.0 mL of Amyl Nitrite to stand in ice
water: no turbidity is produced.
(3) Aldehyde—To 3 mL of a mixture of equal volumes
of silver nitrate TS and aldehyde free-ethanol add ammonia
TS dropwise until the precipitate first formed is redissolved.
JP XVII Official Monographs / Aprindine Hydrochloride 437
Antipyrine Aprindine Hydrochloride

Phenazone アプリンジン塩酸塩
アンチピリン
C22H30N2.HCl: 358.95
C11H12N2O: 188.23
N-(2,3-Dihydro-1H-inden-2-yl)-N?, N?-diethyl-
1,5-Dimethyl-2-phenyl-1,2-dihydro-3H-pyrazol-3-one
N-phenylpropane-1,3-diamine monohydrochloride
[60-80-0]
[33237-74-0]
Antipyrine, when dried, contains not less than
Aprindine Hydrochloride, when dried, contains
99.0z of antipyrine (C11H12N2O).
Description Antipyrine occurs as colorless or white crys- aprindine hydrochloride (C22H30N2.HCl).
tals, or a white, crystalline powder. It is odorless, and has a
Description Aprindine Hydrochloride occurs as a white to
slightly bitter taste.
pale yellowish white crystalline powder. It has a bitter taste,
It is very soluble in water, freely soluble in ethanol (95),
numbing the tongue.
and sparingly soluble in diethyl ether.
It is very soluble in water, in methanol and in acetic acid
A solution of Antipyrine (1 in 10) is neutral.
(100), and freely soluble in ethanol (99.5).
Identification (1) To 5 mL of a solution of Antipyrine It gradually turns brown on exposure to light.
(1 in 100) add 2 drops of sodium nitrite TS and 1 mL of
Identification (1) Dissolve 10 mg of Aprindine Hydro-
dilute sulfuric acid: a deep green color develops.
chloride in a solution of hydrochloric acid in diluted ethanol
(2) To 2 mL of a solution of Antipyrine (1 in 100) add 4
(1 in 2) (1 in 125) to make 50 mL. Determine the absorption
drops of dilute iron (III) chloride TS: a yellow-red color de-
spectrum of this solution as directed under Ultraviolet-
velops. Then add 10 drops of dilute sulfuric acid: the color
changes to light yellow.
(3) To 5 mL of a solution of Antipyrine (1 in 100) add 2
to 3 drops of tannic acid TS: a white precipitate is produced.
(4) To 0.1 g of Antipyrine add 0.1 g of vanillin, 5 mL of
Aprindine Hydrochloride as directed in the potassium chlo-
water and 2 mL of sulfuric acid, boil the mixture, and cool:
a yellow-red precipitate is produced.
Melting point <2.60> 111 – 1139C both spectra exhibit similar intensities of absorption at the
same wave numbers.
Purity (1) Chloride <1.03>—Perform the test with 1.0 g of
(3) To 5 mL of a solution of Aprindine Hydrochloride
Antipyrine. Prepare the control solution with 0.40 mL of
(1 in 50) add 1 mL of dilute nitric acid: this solution re-
sponds to the Qualitative Tests <1.09> for chloride.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Antipy-
rine according to Method 1, and perform the test. Prepare pH <2.54> Dissolve 1.0 g of Aprindine Hydrochloride in 50
the control solution with 2.0 mL of Standard Lead Solution mL of water: the pH of the solution is between 6.4 and 7.0.
(3) Readily carbonizable substances<1.15>—Perform the
test with 0.5 g of Antipyrine: the solution remains colorless. Purity (1) Clarity and color of solution—Dissolve 1.0 g
of Aprindine Hydrochloride in 10 mL of methanol: the solu-
Loss on drying <2.41> Not more than 0.5z (1 g, silica gel,
tion is clear, and its absorbance at 420 nm determined as di-
4 hours).
rected under Ultraviolet-visible Spectrophotometry <2.24> is
Residue on ignition <2.44> Not more than 0.1z (1 g). not more than 0.10.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Aprin-
Assay Dissolve about 0.2 g of Antipyrine, previously dried
and accurately weighed, in 20 mL of sodium acetate TS, add
exactly 30 mL of 0.05 mol/L iodine VS, and allow to stand
for 20 minutes with occasional shaking. Dissolve the precipi-
(3) Related substances—Dissolve 25 mg of Aprindine
tate in 10 mL of chloroform, and titrate <2.50> the excess
iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 3
mL of starch TS). Perform a blank determination.
Each mL of 0.05 mol/L iodine VS use this solution as the standard solution. Perform the test
＝ 9.412 mg of C11H12N2O with exactly 10 mL each of the sample solution and standard
according to the following conditions. Determine each peak
ers.
area of both solutions by the automatic integration method:
the area of the peak other than aprindine obtained from the
sample solution is not larger than 1/10 times the peak area
438 Aprindine Hydrochloride Capsules / Official Monographs JP XVII
of aprindine obtained from the standard solution. Uniformity of dosage units <6.02> Perform the test accord-
Operating conditions— ing to the following method: it meets the requirement of the
Detector: An ultraviolet absorption photometer (wave- Content uniformity test.
length: 254 nm). Take out the contents of 1 capsule of Aprindine Hydro-
Column: A stainless steel column 4.6 mm in inside diame- chloride Capsules, add 30 mL of a solution of hydrochloric
ter and 15 cm in length, packed with octadecylsilanized silica acid in diluted ethanol (1 in 2) (1 in 125), shake vigorously
gel for liquid chromatography (5 mm in particle diameter). for 20 minutes, add a solution of hydrochloric acid in diluted
Column temperature: A constant temperature of about ethanol (1 in 2) (1 in 125) to make exactly V mL so that each
409 C. mL contains about 0.2 mg of aprindine hydrochloride
Mobile phase: Dissolve 3.40 g of potassium dihydrogen (C22H30N2.HCl), and filter. Discard the first 5 mL of the fil-
phosphate in 500 mL of water, and adjust the pH to 3.0 with trate, and use the subsequent filtrate as the sample solution.
hydrochloric acid. To 500 mL of this solution add 500 mL of Proceed as directed in the Assay.
acetonitrile.
Amount (mg) of aprindine hydrochloride (C22H30N2.HCl)
Flow rate: Adjust so that the retention time of aprindine is
＝ MS × AT/AS × V/250
about 6 minutes.
Time span of measurement: About 4 times as long as the MS: Amount (mg) of aprindine hydrochloride for assay
retention time of aprindine. taken
sinker, using 900 mL of water as the dissolution medium, the
Confirm that the peak area of aprindine obtained from 10
dissolution rate in 15 minutes of Aprindine Hydrochloride
mL of this solution is equivalent to 7 to 13z of that obtained
Capsules is not less than 80z.
from 10 mL of the standard solution.
Start the test with 1 capsule of Aprindine Hydrochloride
Capsules, withdraw not less than 20 mL of the medium at
the specified minute after starting the test, and filter through
a membrane filter with a pore size not exceeding 0.45 mm.
factor of the peak of aprindine are not less than 3000 and
Discard the first 10 mL of the filtrate, pipet V mL of the
subsequent filtrate, add water to make exactly V? mL so that
each mL contains about 11 mg of aprindine hydrochloride
(C22H30N2.HCl), and use this solution as the sample solu-
tion. Separately, weigh accurately about 28 mg of aprindine
area of aprindine is not more than 1.5z.
hydrochloride for assay, previously dried in vacuum at 609C
Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- for 4 hours, and dissolve in water to make exactly 100 mL.
um, 609C, 4 hours). Pipet 2 mL of this solution, add water to make exactly 50
Assay Weigh accurately about 0.5 g of Aprindine Hydro- standard solution as directed under Liquid Chromatography
chloride, previously dried, dissolve in 80 mL of acetic acid <2.01> according to the following conditions, and determine
(100), and titrate <2.50> with 0.1 mol/L perchloric acid VS the peak areas, AT and AS, of aprindine in each solution.
(potentiometric titration). Perform a blank determination in
the same manner, and make any necessary correction.
of aprindine hydrochloride (C22H30N2.HCl)
Each mL of 0.1 mol/L perchloric acid VS ＝ MS × AT/AS × V?/V × 1/C × 36
＝ 35.90 mg of C22H30N2.HCl
MS: Amount (mg) of aprindine hydrochloride for assay
Containers and storage Containers—Well-closed contain- taken
ers. C: Labeled amount (mg) of aprindine hydrochloride
Storage—Light-resistant. (C22H30N2.HCl) in 1 capsule
Aprindine Hydrochloride Capsules length: 254 nm).
アプリンジン塩酸塩カプセル
Aprindine Hydrochloride Capsules contain not less Column temperature: A constant temperature of about
than 95.0z and not more than 105.0z of the labeled 409C.
amount of aprindine hydrochloride (C22H30N2.HCl: Mobile phase: Dissolve 3.40 g of potassium dihydrogen
358.95). phosphate in 500 mL of water, and adjust the pH to 3.0 with
hydrochloric acid. To 500 mL of this solution add 500 mL of
acetonitrile.
sules, with Aprindine Hydrochloride.
Flow rate: Adjust so that the retention time of aprindine is
Identification Determine the absorption spectrum of the about 6 minutes.
sample solution obtained in the Assay as directed under System suitability—
Ultraviolet-visible Spectrophotometry <2.24>, it exhibits System performance: When the procedure is run with 20
maxima between 264 nm and 268 nm, and between 271 nm mL of the standard solution under the above operating con-
and 275 nm. ditions, the number of theoretical plates and the symmetry
factor of the peak of aprindine are not less than 3000 and
JP XVII Official Monographs / Arbekacin Sulfate 439
not more than 2.0, respectively. pressed as mass (potency) of arbekacin (C22H44N6O10:
System repeatability: When the test is repeated 6 times 552.62).
Description Arbekacin Sulfate occurs as a white powder.
It is very soluble in water, and practically insoluble in
area of aprindine is not more than 1.5z.
ethanol (99.5).
Assay Take out the contents of not less than 20 Aprindine
Identification (1) Dissolve 10 mg each of Arbekacin Sul-
Hydrochloride Capsules, weigh accurately the mass of the
fate and Arbekacin Sulfate RS in 1 mL of water, and use
contents, and powder. Weigh accurately a portion of the
these solutions as the sample solution and standard solution.
powder, equivalent to about 0.1 g of aprindine hydrochlo-
ride (C22H30N2.HCl), add 60 mL of a solution of hydrochlo-
ric acid in diluted ethanol (1 in 2) (1 in 125), shake vigorously
solution and standard solution on a plate of silica gel for
for 20 minutes, and add a solution of hydrochloric acid in
diluted ethanol (1 in 2) (1 in 125) to make exactly 100 mL.
of ammonia solution (28), methanol, chloroform and
Pipet 10 mL of this solution, add a solution of hydrochloric
ethanol (95) (7:6:4:1) to a distance of about 10 cm, and air-
acid in diluted ethanol (1 in 2) (1 in 125) to make exactly 50
dry the plate. Spray evenly 0.2z ninhydrin-water saturated
mL, and filter, Discard the first 5 mL of the filtrate, and use
1-butanol TS on the plate, and heat at 1009C for 10 minutes:
the subsequent filtrate as the sample solution. Separately,
the principal spot obtained from the sample solution and the
weigh accurately about 50 mg of aprindine hydrochloride for
spot obtained from the standard solution are purple-brown
assay, previously dried in vacuum at 609C for 4 hours, and
in color and their R f values are the same.
dissolve in a solution of hydrochloric acid in diluted ethanol
(2) A solution of Arbekacin Sulfate (1 in 50) responds to
(1 in 2) (1 in 125) to make exactly 50 mL. Pipet 10 mL of this
the Qualitative Tests <1.09> (1) for sulfate.
solution, add a solution of hydrochloric acid in diluted
ethanol (1 in 2) (1 in 125) to make exactly 50 mL, and use Optical rotation <2.49> [a]20
D : ＋69 – ＋799 (0.25 g after
this solution as the standard solution. Determine the absor- drying, water, 25 mL, 100 mm).
bances, AT and AS, of the sample solution and standard so-
lution at 265 nm as directed under Ultraviolet-visible Spec-
0.75 g of Arbekacin Sulfate in 10 mL of water is between 6.0
and 8.0.
Amount (mg) of aprindine hydrochloride (C22H30N2.HCl)
Purity (1) Clarity and color of solution—A solution
＝ M S × AT / AS × 2
obtained by dissolving 1.0 g of Arbekacin Sulfate in 5 mL of
MS: Amount (mg) of aprindine hydrochloride for assay water is clear and colorless.
taken (2) Heavy metals <1.07>—Proceed with 2.0 g of Arbeka-
cin Sulfate according to Method 1, and perform the test.
(3) Dibekacin—Weigh accurately about 20 mg of Ar-
bekacin Sulfate, add exactly 10 mL of the internal standard
Arbekacin Sulfate solution to dissolve, add water to make 20 mL, and use this
アルベカシン硫酸塩
an amount of Dibekacin Sulfate RS, equivalent to about 10
mg (potency), and dissolve in water to make exactly 50 mL.
nal standard solution and water to make 20 mL, and use this
solution as the standard solution. Perform the test with 5 mL
the peak area of dibekacin to that of the internal standard.
Calculate the amount of dibekacin by the following equa-
tion: not more than 2.0z.
Amount (z) of dibekacin
＝ MS/MT × QT/QS × 1/10 × 100
MS: Amount [mg (potency)] of Dibekacin Sulfate RS
taken
C22H44N6O10.xH2SO4 (x ＝ 2 － 2 1/2 )
MT: Amount (mg) of Arbekacin Sulfate taken
3-Amino-3-deoxy-a-D-glucopyranosyl-(1→6)-
[2,6-diamino-2,3,4,6-tetradeoxy-a-D-erythro- Internal standard solution—A solution of bekanamycin sul-
hexopyranosyl-(1→4)]-1-N-[(2S)-4-amino-2- fate (1 in 2000).
hydroxybutanoyl]-2-deoxy-D-streptamine sulfate Operating conditions—
[51025-85-5, Arbekacin] Detector: Fluorometric detector (excitation wavelength:
340 nm, detection wavelength: 460 nm).
Arbekacin Sulfate is the sulfate of a derivative of Column: A stainless steel column 4.6 mm in inside diame-
dibekacin. ter and 15 cm in length, packed with octadecylsilanized silica
It contains not less than 670 mg (potency) and not gel for liquid chromatography (5 mm in particle diameter).
more than 750 mg (potency) per mg, calculated on the Column temperature: A constant temperature of about
dried basis. The potency of Arbekacin Sulfate is ex- 409C.
440 Arbekacin Sulfate Injection / Official Monographs JP XVII
Reaction coil: A column about 0.3 mm in inside diameter method as directed under Microbial Assay for Antibiotics
and about 3 m in length. <4.02> according to the following conditions.
Reaction coil temperature: A constant temperature of (i) Test organism—Bacillus subtilis ATCC 6633
about 509 C. (ii) Culture medium—Use the medium i in 1) under (1)
Mobile phase: Dissolve 8.70 g of sodium 1-pentane sul- Agar media for seed and base layer, having pH 7.8 – 8.0
fonate and 8.52 g of anhydrous sodium sulfate in 980 mL of after sterilization.
water, adjust the pH to 4.0 with acetic acid (100), and add (iii) Standard solutions—Weigh accurately an amount of
water to make 1000 mL. To 230 mL of this solution add 20 Arbekacin Sulfate RS, previously dried, equivalent to about
mL of methanol. 20 mg (potency), dissolve in diluted phosphate buffer solu-
Reagent: Dissolve 12.36 g of boric acid in 960 mL of tion (pH 6.0) (1 in 2) to make exactly 50 mL, and use this so-
water, add 10 mL of a solution of o-phthalaldehyde in lution as the standard stock solution. Keep the standard
ethanol (99.5) (1 in 25), adjust the pH to 10.5 with 8 mol/L stock solution at 5 to 159C and use within 30 days. Take ex-
potassium hydroxide TS, and add water to make 1000 mL. actly a suitable amount of the standard stock solution before
To this solution add 1 mL of 2-mercaptoethanol. use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to
Reaction temperature: A constant temperature of about make solutions so that each mL contains 20 mg (potency) and
509 C. 5 mg (potency), and use these solutions as the high concentra-
Flow rate of mobile phase: 0.5 mL per minute. tion standard solution and low concentration standard solu-
Flow rate of reagent: 1 mL per minute. tion, respectively.
System suitability— (iv) Sample solutions—Weigh accurately an amount of
System performance: Dissolve 20 mg each of Arbekacin Arbekacin Sulfate, equivalent to about 20 mg (potency),
Sulfate, becanamycin sulfate and dibekacin sulfate in 200 and dissolve in water to make exactly 50 mL. Take exactly a
mL of water. When the procedure is run with 5 mL of this so- suitable amount of this solution, add 0.1 mol/L phosphate
lution under the above operating conditions, becanamycin, buffer solution (pH 8.0) to make solutions so that each mL
arbekacin and dibekacin are eluted in this order, and the contains 20 mg (potency) and 5 mg (potency), and use these
resolution between the peaks, becanamycin and arbekacin is solutions as the high concentration sample solution and low
not less than 5 and arbekacin and dibekacin is not less than concentration sample solution, respectively.
1.5, respectively.
conditions, the relative standard deviation of the ratio of the
peak area of dibekacin to that of the internal standard is not Arbekacin Sulfate Injection
more than 2.0z.
アルベカシン硫酸塩注射液
(4) Related substances—Dissolve 20 mg of Arbekacin
Sulfate in 20 mL of water, and use this solution as the sam-
ple solution. Pipet 3 mL of the sample solution, add water to Arbekacin Sulfate Injection is an aqueous injection.
make exactly 250 mL, and use this solution as the standard It contains not less than 90.0z and not more than
solution. Perform the test with exactly 5 mL each of the sam- 110.0z of the labeled potency of arbekacin sulfate
ple solution and standard solution as directed under Liquid (C22H44N6O10: 552.62).
tions, and determine the area of each peak by the automatic
tions, with Arbekacin Sulfate.
integration method: the total area of the peaks other than ar-
bekacin and dibekacin obtained from the sample solution is Description Arbekacin Sulfate Injection occurs as a clear
not larger than the peak area of arbekacin obtained from the and colorless liquid.
standard solution.
Identification To 0.2 mL of Arbekacin Sulfate Injection
add 1 mL of water, and use this solution as the sample solu-
Detector, column, column temperature, reaction coil,
tion. Separately, dissolve 10 mg of Arbekacin Sulfate RS in 1
reaction coil temperature, mobile phase, reagent, reaction
mL of water, and use this solution as the standard solution.
temperature, flow rate of mobile phase, and flow rate of rea-
gent: Proceed as directed in the operating conditions in the
Purity (3).
solution and standard solution on a plate of silica gel for
Time span of measurement: About 1.5 times as long as the
thin-layer chromatography. Develop with a mixture of am-
retention time of arbekacin.
monia solution (28), methanol, chloroform and ethanol (95)
(7:6:4:1) to a distance of about 12 cm, and air-dry the plate.
System performance: Dissolve 10 mg each of Arbekacin
Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS
Sulfate and dibekacin sulfate in 200 mL of water. When the
on the plate, and heat at 809C for 10 minutes: the principal
spot with the sample solution and the spot with the standard
operating conditions, arbekacin and dibekacin are eluted in
solution show a purple-brown color and the same R f value.
less than 1.5. Osmotic pressure ratio <2.47> 0.8 – 1.2 (for the preparation
System repeatability: When the test is repeated 6 times intended for intramuscular use).
pH <2.54> 6.0 – 8.0
of arbekacin is not more than 5.0z. Bacterial endotoxins <4.01> Less than 0.50 EU/mg (po-
tency).
Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced
pressure not exceeding 0.67 kPa, 609C, 3 hours). Extractable volume <6.05> It meets the requirement.
Assay Perform the test according to the Cylinder-plate Foreign insoluble matter <6.06> Perform the test according
JP XVII Official Monographs / Argatroban Hydrate 441
to Method 1: it meets the requirement. Optical rotation <2.49> [a]20

D : ＋175 – ＋1859(0.2 g calcu-
lated on the anhydrous basis, methanol, 25 mL, 100 mm).
ment. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
Argatroban Hydrate according to Method 2, and perform
Assay Perform the test according to the Cylinder-plate (2) Arsenic <1.11>—Incinerate 2.0 g of Argatroban Hy-
method as directed under Microbial Assay for Antibiotics drate according to Method 4. After cooling, add 10 mL of
<4.02> according to the following conditions. dilute hydrochloric acid to the residue, dissolve by warming
(i) Test organism, Culture medium and Standard solu- on a water bath, and perform the test using this solution as
tions: Proceed as directed in the Assay under Arbekacin Sul- the test solution. Add 10 mL of a solution of magnesium ni-
fate. trate hexahydrate in ethanol (95) (1 in 10), then add 1.5 mL
(ii) Sample solutions—Take exactly a volume of Arbeka- of hydrogen peroxide (30), and fire to burn (not more than 1
cin Sulfate Injection, equivalent to about 20 mg (potency), ppm).
and add water to make exactly 50 mL. Take exactly a suita- (3) Related substance 1—Dissolve 50 mg of Argatroban
ble amount of this solution, add 0.1 mol/L phosphate buffer Hydrate in 40 mL of methanol, add water to make 100 mL,
solution (pH 8.0) to make solutions so that each mL contains and use this solution as the sample solution. Perform the test
20 mg (potency) and 5 mg (potency), and use these solutions with 10 mL of the sample solution as directed under Liquid
as the high concentration sample solution and low concen- Chromatography <2.01> according to the following condi-
tration sample solution, respectively. tions. Determine each peak area from the sample solution by
the automatic integration method, and calculate the amount
of them by the area percentage method: the amount of each
peak other than argatroban is not more than 0.1z.
Argatroban Hydrate Detector: An ultraviolet absorption photometer (wave-
length: 254 nm).
アルガトロバン水和物
459C.
Mobile phase A: To 2.5 mL of acetic acid (100) add water
to make 1000 mL, and adjust the pH to 5.0 with ammonia
TS. To 500 mL of this solution add 500 mL of methanol.
Mobile phase B: To 2.5 mL of acetic acid (100) add water
to make 1000 mL, and adjust the pH to 5.0 with ammonia
C23H36N6O5S.H2O: 526.65 TS. To 200 mL of this solution add 800 mL of methanol.
(2R,4R)-4-Methyl-1-((2S)-2-{[(3RS)-3-methyl- Flowing of mobile phase: Control the gradient by mixing
1,2,3,4-tetrahydroquinolin-8-yl]sulfonyl}amino- the mobile phases A and B as directed in the following table.
5-guanidinopentanoyl)piperidine-2-carboxylic acid
monohydrate Time after injection Mobile phase A Mobile phase B
[141396-28-3] of sample (min) (volz) (volz)
Argatroban Hydrate contains not less than 0–5 100 0

98.5z and not more than 101.0z of argatroban 5 – 35 100 → 5 0 → 95
(C23H36N6O5S: 508.63), calculated on the anhydrous
basis. Flow rate: About 1.0 mL per minute.
Description Argatroban Hydrate occurs as white, crystals Time span of measurement: About 1.5 times as long as the
or crystalline powder. It has a bitter taste. retention time of argatroban, beginning after the solvent
It is freely soluble in acetic acid (100), sparingly soluble in peak.
methanol, slightly soluble in ethanol (99.5), and very slightly System suitability—
soluble in water. Test for required detectability: To 1 mL of the sample so-
It is gradually decomposed on exposure to light. lution add the mobile phase A to make 100 mL, and use this
Identification (1) Determine the absorption spectrum of a mL of the solution for system suitability test, and add the
solution of Argatroban Hydrate in ethanol (99.5) (1 in mobile phase A to make exactly 10 mL. Confirm that the
20,000) as directed under Ultraviolet-visible Spectropho- peak area of argatroban obtained from 10 mL of this solu-
tometry <2.24>, and compare the spectrum with the Refer- tion is equivalent to 7 to 13z of that obtained from 10 mL of
ence Spectrum: both spectra exhibit similar intensities of ab- the solution for system suitability test.
sorption at the same wavelengths. System performance: Dissolve 5 mg of Argatroban Hy-
(2) Determine the infrared absorption spectrum of Ar- drate and 5 mL of methyl benzoate in 40 mL of methanol,
gatroban Hydrate as directed in the potassium bromide disk and add water to make 100 mL. To 5 mL of this solution
method under Infrared Spectrophotometry <2.25>, and com- add 40 mL of methanol and water to make 100 mL. When
pare the spectrum with the Reference Spectrum: both spectra the procedure is run with 10 mL of this solution under the
exhibit similar intensities of absorption at the same wave above operating conditions, methyl benzoate and argatroban
numbers. are eluted in this order with the resolution between these
442 L-Arginine / Official Monographs JP XVII
peaks being not less than 3. Each mL of 0.1 mol/L perchloric acid VS
System repeatability: When the test is repeated 6 times ＝ 50.86 mg of C23H36N6O5S
tion of the peak area of argatroban is not more than 2.0z.
(4) Related substance 2—Dissolve 0.10 g of Argatroban
Hydrate in 10 mL of methanol, and use this solution as the
sample solution. Pipet 3 mL of the sample solution, and add L-Arginine
methanol to make exactly 100 mL. Pipet 5 mL of this solu-
L-アルギニン
tion, add methanol to make exactly 50 mL, and use this solu-
tion as the standard solution. Perform the test with these so-
lutions as directed under Thin-layer Chromatography <2.03>.
Spot 10 mL each of the sample solution and standard solu-
tion on a plate of silica gel with fluorescent indicator for
thin-layer chromatography. Develop the plate with a mixture C6H14N4O2: 174.20
of methanol, ethyl acetate and water (10:10:1) to a distance (2S )-2-Amino-5-guanidinopentanoic acid
of about 10 cm, and air-dry the plate. Examine under ultra- [74-79-3]
violet light (main wavelength: 254 nm): the number of spots
other than the principal spot obtained from the sample solu- L-Arginine, when dried, contains not less than
tion is not more than 2, and they are not more intense than 98.5z and not more than 101.0z of L-arginine
the spot obtained from the standard solution. (C6H14N4O2).
Water <2.48> 2.5 – 4.5z (0.2 g, volumetric titration, direct Description L-Arginine occurs as white, crystals or crystal-
titration). line powder. It has a characteristic odor.
It is freely soluble in water and in formic acid, and practi-
cally insoluble in ethanol (99.5).
Isomer ratio Dissolve 50 mg of Argatroban Hydrate in 50 It dissolves in dilute hydrochloric acid.
mL of methanol, add water to make 100 mL, and use this so- It is hygroscopic.
lution as the sample solution. Perform the test with 10 mL of
the sample solution as directed under Liquid Chromatogra-
of previously dried L-Arginine as directed in the potassium
phy <2.01> according to the following conditions. Determine
the areas of two adjacent peaks, Aa and Ab, having the reten-
tion times of about 40 minutes, where Aa is the peak area of
shorter retention time and Ab is the peak area of longer
retention time: Ab/(Aa ＋ Ab) is between 0.30 and 0.40.
Operating conditions— Optical rotation <2.49> [a]20
D : ＋26.9 – ＋27.99(after dry-
Detector: An ultraviolet absorption photometer (wave- ing, 2 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm).
length: 254 nm).
pH <2.54> The pH of a solution prepared by dissolving
1.0 g of L-Arginine in 10 mL of water is between 10.5 and
12.0.
Column temperature: A constant temperature of about Purity (1) Clarity and color of solution—A solution ob-
409 C. tained by dissolving 1.0 g of L-Arginine in 10 mL of water is
Mobile phase: To 500 mL of water add 500 mL of metha- clear and colorless.
nol, 13 mL of diluted 40z tetrabutylammonium hydroxide (2) Chloride <1.03>—Perform the test with 0.5 g of
TS (1 in 4) and 0.68 mL of phosphoric acid, and adjust the L-Arginine. Prepare the control solution with 0.30 mL of
pH to 6.8 with ammonia TS and diluted ammonia solution 0.01 mol/L hydrochloric acid VS (not more than 0.021z).
(28) (1 in 20). (3) Sulfate <1.14>—Perform the test with 0.6 g of
Flow rate: Adjust so that the retention time of the peak L-Arginine. Prepare the control solution with 0.35 mL of
having the shorter retention time of the two peaks of ar- 0.005 mol/L sulfuric acid VS (not more than 0.028z).
gatroban is about 40 minutes. (4) Ammonium <1.02>—Perform the test with 0.25 g of
System suitability— L-Arginine, using the distillation under reduced pressure.
System performance: When the procedure is run with 10 Prepare the control solution with 5.0 mL of Standard Am-
mL of the sample solution under the above operating condi- monium Solution (not more than 0.02z).
tions, the resolution between the two peaks is not less than (5) Heavy metals <1.07>—Dissolve 2.0 g of L-Arginine in
1.2. 30 mL of water, add 1 drop of phenolphthalein TS, neutral-
System repeatability: When the test is repeated 6 times ize with dilute hydrochloric acid, add 2 mL of dilute acetic
with 10 mL of the sample solution under the above operating acid and water to make 50 mL, and perform the test. Pre-
conditions, the relative standard deviation of the total area pare the control solution with 2.0 mL of Standard Lead So-
of the two separate peaks of argatroban is not more than lution (not more than 10 ppm).
2.0z. (6) Iron <1.10>—Prepare the test solution with 1.0 g of
L-Arginine according to Method 1, and perform the test
Assay Weigh accurately about 0.5 g of Argatroban Hy-
drate, dissolve in 20 mL of acetic acid for nonaqueous titra-
tion, add 40 mL of acetone for nonaqueous titration, and
(7) Related substances—Dissolve 0.10 g of L-Arginine in
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-
10 mL of water, and use this solution as the sample solution.
Pipet 1 mL of the sample solution, and add water to make
JP XVII Official Monographs / L-Arginine Hydrochloride Injection 443
exactly 50 mL. Pipet 2 mL of this solution, add water to (2) Sulfate <1.14>—Perform the test with 0.6 g of
make exactly 20 mL, and use this solution as the standard L-Arginine Hydrochloride. Prepare the control solution with
solution. Perform the test with these solutions as directed 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than
under Thin-layer Chromatography <2.03>. Spot 5 mL each of 0.028z).
the sample solution and standard solution on a plate of silica (3) Ammonium <1.02>—Perform the test with 0.25 g of
gel for thin-layer chromatography. Develop the plate with a L-Arginine Hydrochloride, using the distillation under
mixture of 2-propanol and ammonia solution (28) (7:3) to a reduced pressure. Prepare the control solution with 5.0 mL
distance of about 10 cm, and dry the plate at 809C for 30 of Standard Ammonium Solution (not more than 0.02z).
minutes. Spray evenly ninhydrin-butanol TS on the plate, (4) Heavy metals <1.07>—Proceed with 1.0 g of
and heat at 809C for 10 minutes: the spot other than the L-Arginine Hydrochloride according to Method 1, and
principal spot with the sample solution is not more intense perform the test. Prepare the control solution with 2.0 mL
than the spot with the standard solution. of Standard Lead Solution (not more than 20 ppm).
C,
of L-Arginine Hydrochloride according to Method 1, and
3 hours).
Residue on ignition <2.44> Not more than 0.1z (1 g). (6) Related substances—Dissolve 0.20 g of L-Arginine
Hydrochloride in 10 mL of water, and use this solution as
Assay Weigh accurately about 80 mg of L-Arginine, previ-
the sample solution. Pipet 1 mL of the sample solution, and
ously dried, dissolve in 3 mL of formic acid, add 50 mL of
add water to make exactly 10 mL. Pipet 1 mL of this solu-
acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric
tion, add water to make exactly 25 mL, and use this solution
nation in the same manner, and make any necessary correc-
tion.
Each mL of 0.1 mol/L perchloric acid VS on a plate of silica gel for thin-layer chromatography.
＝ 8.710 mg of C6H14N4O2 Develop the plate with a mixture of ethanol (99.5), water, 1-
butanol and ammonia water (28) (2:1:1:1) to a distance of
about 10 cm, and dry the plate at 1009 C for 30 minutes.
Spray evenly a solution of ninhydrin in acetone (1 in 50) on
the plate, and heat at 809C for 5 minutes: the spots other
L-Arginine Hydrochloride than the principal spot from the sample solution are not
more intense than the spot from the standard solution.
L-アルギニン塩酸塩
3 hours).
Assay Weigh accurately about 0.1 g of L-Arginine Hydro-
C6H14N4O2.HCl: 210.66 chloride, previously dried, dissolve in 2 mL of formic acid,
(2S )-2-Amino-5-guanidinopentanoic acid add exactly 15 mL of 0.1 mol/L perchloric acid VS, and heat
monohydrochloride on a water bath for 30 minutes. After cooling, add 45 mL of
[1119-34-2] acetic acid (100), and titrate <2.50> the excess perchloric acid
with 0.1 mol/L sodium acetate VS (potentiometric titration).
L-Arginine Hydrochloride, when dried, contains Perform a blank determination.
not less than 98.5z of L-arginine hydrochloride
(C6H14N4O2.HCl). ＝ 10.53 mg of C6H14N4O2.HCl
Description L-Arginine Hydrochloride occurs as white,
crystals or crystalline powder. It is odorless, and has a slight,
characteristic taste.
It is freely soluble in water and in formic acid, and very
slightly soluble in ethanol (95). L-Arginine Hydrochloride Injection
Identification (1) Determine the infrared absorption L-アルギニン塩酸塩注射液
spectrum of L-Arginine Hydrochloride, previously dried, as
directed in the potassium bromide disk method under In-
L-Arginine Hydrochloride Injection is an aqueous
frared Spectrophotometry <2.25>, and compare the spectrum
injection.
with the Reference Spectrum: both spectra exhibit similar
It contains not less than 9.5 w/vz and not
intensities of absorption at the same wave numbers.
more than 10.5 w/vz of L-arginine hydrochloride
(2) A solution of L-Arginine Hydrochloride (1 in 50)
(C6H14N4O2.HCl: 210.66).
D : ＋21.5 – ＋23.59(after dry-
ing, 2 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). L-Arginine Hydrochloride 100 g
Water for Injection or Sterile Water
pH <2.54> Dissolve 1.0 g of L-Arginine Hydrochloride in 10
for Injection in Containers a sufficient quantity
mL of water: the pH of this solution is between 4.7 and 6.2.
To make 1000 mL
of L-Arginine Hydrochloride in 10 mL of water: the solution Prepare as directed under Injections, with the above
is clear and colorless. ingredients.
444 Arotinolol Hydrochloride / Official Monographs JP XVII
No preservative is added. (2) Determine the infrared absorption spectrum of
Arotinolol Hydrochloride, previously dried, as directed in
Description L-Arginine Hydrochloride Injection is a clear,
colorless liquid.
Identification (1) To 5 mL of a solution of L-Arginine erence Spectrum: both spectra exhibit similar intensities of
Hydrochloride Injection (1 in 100) add 1 mL of ninhydrin absorption at the same wave numbers.
TS, and heat for 3 minutes: a blue-purple color develops. (3) A solution of Arotinolol Hydrochloride (1 in 200)
(2) To 5 mL of a solution of L-Arginine Hydrochloride responds to the Qualitative Tests <1.09> (2) for chloride.
Injection (1 in 10) add 2 mL of sodium hydroxide TS and 1
to 2 drops of a solution of 1-naphthol in ethanol (95) (1 in
Arotinolol Hydrochloride according to Method 2, and per-
1000), allow to stand for 5 minutes, and add 1 to 2 drops of
sodium hypochlorite TS: a red-orange color develops.
pH <2.54> 5.0 – 6.0 (2) Related substances—Dissolve 0.05 g of Arotinolol
Hydrochloride in 10 mL of methanol, and use this solution
Bacterial endotoxins <4.01> Less than 0.50 EU/mL.
Extractable volume <6.05> It meets the requirement. add methanol to make exactly 100 mL. Pipet 1 mL of this
solution, add methanol to make exactly 10 mL, and use this
Insoluble particulate matter <6.07> It meets the require- <2.03>. Spot 40 mL each of the sample solution and standard
ment. solution on a plate of silica gel with fluorescent indicator for
of chloroform, methanol, acetone and ammonia solution
(28) (30:10:10:1) to a distance of about 12 cm, and air-dry
Assay Pipet 20 mL of L-Arginine Hydrochloride Injection, the plate. Examine under ultraviolet light (main wavelength:
add 7.5 mol/L hydrochloric acid TS to make exactly 100 254 nm): the spots other than the principal spot from the
mL, and determine the optical rotation aD as directed under sample solution are not more intense than the spot from the
Optical Rotation Determination <2.49> at 20 ± 19C in a standard solution.
100-mm cell.
Amount (mg) of L-arginine hydrochloride (C6H14N4O2.HCl) um, 1059 C, 4 hours).
＝ aD × 4444
Assay Weigh accurately about 1.5 g of Arotinolol Hydro-
chloride, previously dried, dissolve in dimethylsulfoxide to
make exactly 25 mL. Pipet 5 mL of this solution, add 100
Arotinolol Hydrochloride mL of water and 5 mL of sodium hydroxide TS, and extract
with three 50-mL portions of dichloromethane. Filter each
アロチノロール塩酸塩
dichloromethane extract through a pledget of absorbent cot-
ton with anhydrous sodium sulfate on it. Evaporate com-
bined filtrate to dryness in vacuum. Dissolve the residue in
70 mL of acetic acid (100), and titrate <2.50> with 0.05
mol/L perchloric acid VS (potentiometric titration). Per-
form a blank determination, and make any necessary correc-
C15H21N3O2S3.HCl: 408.00
tion.
5-{2-[(2RS )-3-(1,1-Dimethylethyl)amino-
2-hydroxypropylsulfanyl]-1,3-thiazol-4-yl}thiophene- Each mL of 0.05 mol/L perchloric acid VS
2-carboxamide monohydrochloride ＝ 20.40 mg of C15H21N3O2S3.HCl
[68377-91-3]
Arotinolol Hydrochloride, when dried, contains
not less than 99.0z of arotinolol hydrochloride
(C15H21N3O2S3.HCl).
Description Arotinolol Hydrochloride occurs as a white to
light yellow crystalline powder.
It is freely soluble in dimethylsulfoxide, slightly soluble in
methanol and in water, very slightly soluble in ethanol
(99.5), and practically insoluble in diethyl ether.
A solution of Arotinolol Hydrochloride in methanol (1 in
125) does not show optical rotation.
solution of Arotinolol Hydrochloride in methanol (1 in
JP XVII Official Monographs / Ascorbic Acid 445
allow to stand in a dark place at room temperature for 45

Arsenical Paste minutes, and titrate <2.50> the liberated iodine with 0.1
mol/L sodium thiosulfate VS (indicator: 5 mL of starch TS).
亜ヒ酸パスタ Perform a blank determination, and make any necessary
correction.
Arsenical Paste contains not less than 36.0z and Each mL of 0.1 mol/L sodium thiosulfate VS
not more than 44.0z of arsenic trioxide (As2O3: ＝ 4.946 mg of As2O3
197.84).
Arsenic Trioxide, finely powdered 40 g
Procaine Hydrochloride, finely Arsenic Trioxide
powdered 10 g
Hydrophilic Cream 30 g
Arsenous Acid
Clove Oil a suitable quantity
三酸化二ヒ素
Medicinal Carbon a suitable quantity
To make 100 g
As2O3: 197.84
Mix Arsenic Trioxide and Procaine Hydrochloride with
Hydrophilic Cream, and add Clove Oil to make a suitably Arsenic Trioxide, when dried, contains not less than
viscous liquid, followed by Medicinal Carbon for coloring. 99.5z of arsenic trioxide (As2O3).
Description Arsenical Paste is grayish black and has the Description Arsenic Trioxide occurs as a white powder.
odor of clove oil. It is odorless. It is practically insoluble in water, in ethanol
Identification (1) Place 0.1 g of Arsenical Paste in a small (95) and in diethyl ether.
flask, add 5 mL of fuming nitric acid and 5 mL of sulfuric It dissolves in sodium hydroxide TS.
acid, and heat over a flame until the reacting liquid becomes Identification Dissolve 0.2 g of Arsenic Trioxide in 40 mL
colorless and white fumes begin to evolve. After cooling, of water by heating on a water bath: the solution responds to
add the reacting liquid to 20 mL of water cautiously, and the Qualitative Tests <1.09> for arsenite.
add 10 mL of hydrogen sulfide TS while warming: a yellow
precipitate is produced (arsenic trioxide). Purity Clarity of solution—To 1.0 g of Arsenic Trioxide
(2) Shake thoroughly 0.5 g of Arsenical Paste with 25 add 10 mL of ammonia TS, and heat gently: the solution is
mL of diethyl ether, 5 mL of dilute hydrochloric acid and 20 clear.
mL of water, separate the water layer, and filter: 5 mL of the Loss on drying <2.41> Not more than 0.5z (1 g, 1059C,
filtrate responds to the Qualitative Tests <1.09> for primary 3 hours).
aromatic amines (procaine hydrochloride).
(3) Shake thoroughly 0.5 g of Arsenical Paste with 25 Assay Weigh accurately about 0.15 g of Arsenic Trioxide,
mL of diethyl ether and 25 mL of water, separate the water previously dried, dissolve in 20 mL of a solution of sodium
layer, filter, and use the filtrate as the sample solution. Dis- hydroxide (1 in 25), by warming, if necessary. Add 40 mL of
solve 0.01 g of procaine hydrochloride in 5 mL of water, and water and 2 drops of methyl orange TS, then add dilute
use this solution as the standard solution. Perform the test hydrochloric acid until the color of the solution becomes
with these solutions as directed under Thin-layer Chroma- light red. Add 2 g of sodium hydrogen carbonate and 50 mL
tography <2.03>. Spot 5 mL each of the sample solution and of water to this solution, and titrate <2.50> with 0.05 mol/L
standard solution on a plate of silica gel with fluorescent in- iodine VS (indicator: 3 mL of starch TS).
dicator for thin-layer chromatography. Develop the plate Each mL of 0.05 mol/L iodine VS ＝ 4.946 mg of As2O3
with a mixture of ethyl acetate, ethanol (99.5) and ammonia
solution (28) (50:5:1) to a distance of about 10 cm, and air- Containers and storage Containers—Tight containers.
dry the plate. Examine under ultraviolet light (main wave-
length: 254 mm): the spots from the sample solution and
standard solution exhibit the same R f value. Ascorbic Acid
Assay Weigh accurately about 0.3 g of Arsenical Paste into Vitamin C
a 150-mL Kjeldahl flask, add 5 mL of fuming nitric acid and
10 mL of sulfuric acid, and shake thoroughly. Heat cau- アスコルビン酸
tiously the mixture, gently at first, and then continue strong
heating, until red fumes of nitrogen oxide are sparingly
evolved. After cooling, add 5 mL of fuming nitric acid, heat
again until red fumes of nitrogen oxide are no longer evolved
and the reacting liquid becomes clear, and cool. Add 30 mL
of a saturated solution of ammonium oxalate monohydrate,
heat again until white fumes of sulfuric acid are evolved, and C6H8O6: 176.12
L-threo-Hex-2-enono-1,4-lactone
continue the heating for 10 minutes. Decompose completely
oxalic acid, cool, transfer cautiously the colorless reacting [50-81-7]
liquid to a glass-stoppered flask, containing 40 mL of water.
Wash thoroughly the Kjeldahl flask with 60 mL of water, Ascorbic Acid, when dried, contains not less than
add the washings to the content of the glass-stoppered flask, 99.0z of L-ascorbic acid (C6H8O6).
and cool. Dissolve 3 g of potassium iodide in this solution, Description Ascorbic Acid occurs as white crystals or a
446 Ascorbic Acid Injection / Official Monographs JP XVII
white crystalline powder. It is odorless, and has an acid equivalent to 5 mg of Ascorbic Acid. Add a solution of
taste. metaphosphoric acid (1 in 50) to make 5 mL, and proceed
It is freely soluble in water, sparingly soluble in ethanol with this solution as directed in the Identification (2) under
(95), and practically insoluble in diethyl ether. Ascorbic Acid.
Melting point: about 1909C (with decomposition). (3) Ascorbic Acid Injection responds to the Qualitative
Tests (1) for sodium salt.
Identification (1) To 5 mL each of a solution of Ascorbic
Acid (1 in 50) add 1 drop of potassium permanganate TS or pH <2.54> 5.6 – 7.4
1 to 2 drops of 2,6-dichloroindophenol sodium TS: the color
of the solution is discharged immediately in each case.
(2) Dissolve 0.1 g of Ascorbic Acid in 100 mL of a solu- Extractable volume <6.05> It meets requirement.
tion of metaphosphoric acid (1 in 50). To 5 mL of the solu-
tion add iodine TS until the color of the solution becomes
light yellow. Then add 1 drop of a solution of copper (II)
sulfate pentahydrate (1 in 1000) and 1 drop of pyrrole, and Insoluble particulate matter <6.07> It meets the require-
warm the mixture at 509C for 5 minutes: a blue color de- ment.
velops.
D : ＋20.5 – ＋21.59 (2.5 g, brane filtration method: it meets the requirement.
water, 25 mL, 100 mm).
Assay Measure exactly a volume of Ascorbic Acid Injec-
pH <2.54> Dissolve 1.0 g of Ascorbic Acid in 20 mL of tion, equivalent to about 0.1 g of L-ascorbic acid (C6H8O6),
water: the pH of this solution is between 2.2 and 2.5. previously diluted with metaphosphoric acid-acetic acid TS,
if necessary, and add metaphosphoric acid-acetic acid TS to
make exactly 200 mL. Measure exactly 2 mL of the solution,
of Ascorbic Acid in 20 mL of water: the solution is clear and
and shake with 8 mL of metaphosphoric acid-acetic acid TS
colorless.
and 2 mL of hydrogen peroxide TS. Titrate <2.50> with 2,6-
(2) Heavy metals <1.07>—Perform the test with 1.0 g of
dichloroindophenol sodium TS for titration until a light red
Ascorbic Acid according to Method 1. Prepare the control
color persists for 5 seconds. Perform a blank determination,
solution with 2.0 mL of Standard Lead Solution (not more
and make any necessary correction.
than 20 ppm).
Each mL of 2, 6-dichlorophenol-indophenol
sodium TS for titration
24 hours).
＝ A mg of C6H8O6
A is decided by the following standardization of 2,6-
Assay Weigh accurately about 0.2 g of Ascorbic Acid, dichloroindophenol sodium TS for titration.
previously dried, dissolve in 50 mL of a solution of meta- 2,6-Dichlorophenol-indophenol sodium TS for titration:
phosphoric acid (1 in 50), and titrate <2.50> with 0.05 mol/L Preparation—Dissolve 42 mg of sodium hydrogen carbon-
iodine VS (indicator: 1 mL of starch TS). ate in 50 mL of water, add 0.5 g of 2,6-dichloroindophenol
sodium dihydrate and water to make 200 mL, and filter. Pre-
Each mL of 0.05 mol/L iodine VS
pare before use.
＝ 8.806 mg of C6H8O6
Standardization—Weigh accurately about 50 mg of Ascor-
Containers and storage Containers—Tight containers. bic Acid RS, previously dried in a desiccator (silica gel) for
Storage—Light-resistant. 24 hours, and dissolve in metaphosphoric acid-acetic acid TS
to make exactly 100 mL. Pipet 2 mL of this solution, shake
with 8 mL of metaphosphoric acid-acetic acid TS and 2 mL
Ascorbic Acid Injection of hydrogen peroxide TS, and titrate <2.50> with 2,6-
dichloroindophenol sodium TS for titration until a light red
Vitamin C Injection color persists for 5 seconds. Perform a blank determination,
and make any necessary correction. Calculate the quantity
アスコルビン酸注射液 (A mg) of L-ascorbic acid (C6H8O6) equivalent to 1 mL of
this test solution.
Ascorbic Acid Injection is an aqueous injection. Containers and storage Containers—Hermetic containers.
It contains not less than 95.0z and not more than Storage—Under nitrogen atmosphere.
115.0z of the labeled amount of L-ascorbic acid
(C6H8O6: 176.12).
Method of preparation Prepare as directed under Injec- Ascorbic Acid Powder
tions, with the sodium salt of Ascorbic Acid.
Vitamin C Powder
Description Ascorbic Acid Injection occurs as a clear, col-
orless liquid. アスコルビン酸散
Identification (1) Measure a volume of Ascorbic Acid
Injection, equivalent to 0.5 g of Ascorbic Acid, and add Ascorbic Acid Powder contains not less than 95.0z
water to make 25 mL. Proceed with 5 mL each of the solu- and not more than 120.0z of the labeled amount of
tion as directed in the Identification (1) under Ascorbic L-ascorbic acid (C6H8O6: 176.12).
Acid.
Method of preparation Prepare as directed under Granules
(2) Measure a volume of Ascorbic Acid Injection,
or Powders, with Ascorbic Acid.
JP XVII Official Monographs / Ascorbic Acid and Calcium Pantothenate Tablets 447
Identification (1) Weigh a portion of Ascorbic Acid Pow- Acid and Calcium Pantothenate Tablets, equivalent to 0.5 g
der, equivalent to 0.5 g of Ascorbic Acid, add 30 mL of of Ascorbic Acid, add 30 mL of water, shake for 1 minute,
water, shake for 1 minute, and filter. Proceed with 5 mL and filter. Proceed as directed in the Identification (1) under
each of the filtrate as directed in the Identification (1) under Ascorbic Acid using 5 mL each of the filtrate.
Ascorbic Acid. (2) To a quantity of powdered Ascorbic Acid and Cal-
(2) Weigh a portion of Ascorbic Acid Powder, equiva- cium Pantothenate Tablets, equivalent to 3 mg of Calcium
lent to about 0.01 g of Ascorbic Acid, add 10 mL of a solu- Pantothenate, add 20 mL of ethanol (95), shake vigorously
tion of metaphosphoric acid (1 in 50), shake for 1 minute, for 10 minutes, centrifuge, and use the supernatant liquid as
and filter. Proceed with 5 mL of the filtrate as directed in the the sample solution. Separately, dissolve 3 mg of calcium
Identification (2) under Ascorbic Acid. pantothenate in 20 mL of ethanol (95), and use this solution
Purity Rancidity—Ascorbic Acid Powder is free from any
unpleasant or rancid odor and taste.
Assay Weigh accurately a portion of Ascorbic Acid Pow- tion on a plate of silica gel for thin-layer chromatography.
der, equivalent to about 0.1 g of L-ascorbic acid (C6H8O6), Develop the plate with a mixture of ethyl acetate, methanol
extract with several successive portions of metaphosphoric and dilute acetic acid (5:3:2) to a distance of about 10 cm,
acid-acetic acid TS, combine the extracts, and filter. Wash and air-dry the plate. Spray evenly a solution of ninhydrin in
the residue with metaphosphoric acid-acetic acid TS. Com- ethanol (95) (1 in 200) on the plate, and heat at 1209C for 20
bine the filtrates and washings, and add metaphosphoric minutes: one of the spot obtained from the sample solution
acid-acetic acid to make exactly 200 mL. Pipet 2 mL of the and the spot obtained from the standard solution are purple
solution, and shake with 8 mL of metaphosphoric acid-acetic in color and their Rf value are the same.
acid TS and 2 mL of hydrogen peroxide TS. Titrate <2.50>
Uniformity of dosage units <6.02> (1) L-Ascorbic acid—
with 2,6-dichloroindophenol sodium TS for titration until a
Perform the Mass variation test, or the Content uniformity
light red color persists for 5 seconds. Perform a blank deter-
test according to the following method: it meets the require-
mination, and make any necessary correction.
ment.
Each mL of 2,6-dichlorophenol-indophenol To 1 tablet of Ascorbic Acid and Calcium Pantothenate
sodium TS for titration Tablets add 100 mL of a solution of metaphosphoric acid (1
＝ A mg of C6H8O6 in 50), stir thoroughly, and titrate <2.50> with 0.05 mol/L
iodine VS (indicator: 1 mL of starch TS).
A is decided by the following standardization of 2,6-
dichloroindophenol sodium TS for titration. Each mL of 0.05 mol/L iodine VS ＝ 8.806 mg of C6H8O6
2,6-Dichlorophenol-indophenol sodium TS for titration:
(2) Calcium pantothenate—Perform the test according
Preparation—Dissolve 42 mg of sodium hydrogen carbon-
to the following method: it meets the requirement of the
ate in 50 mL of water, add 0.05 g of 2,6-dichloroindophenol
sodium dihydrate and water to make 200 mL, and filter. Pre-
To 1 tablet of Ascorbic Acid and Calcium Pantothenate
pare before use.
Tablets add exactly V mL of the internal standard solution
Standardization—Weigh accurately about 50 mg of Ascor-
so that each mL contains about 0.15 mg of calcium panto-
bic Acid RS, previously dried in a desiccator (silica gel) for
thenate (C18H32CaN2O10), shake vigorously for 15 minutes,
24 hours, and dissolve in metaphosphoric acid-acetic acid TS
filter through a membrane filter with a pore size not exceed-
to make exactly 100 mL. Pipet 2 mL of this solution, shake
ing 0.45 mm, and use the filtrate as the sample solution.
with 8 mL of metaphosphoric acid-acetic acid TS and 2 mL
Then, proceed as directed in the Assay (2).
of hydrogen peroxide TS, and titrate <2.50> with 2,6-
dichloroindophenol sodium TS for titration until a light red Amount (mg) of calcium pantothenate (C18H32CaN2O10)
color persists for 5 seconds. Perform a blank determination, ＝ MS × QT/QS × V/200
and make any necessary correction. Calculate the quantity
MS: Amount (mg) of Calcium Pantothenate RS taken, cal-
(A mg) of L-ascorbic acid (C6H8O6) equivalent to 1 mL of
this test solution.
Internal Standard Solution—A solution of acetaminophen
(1 in 50,000).
Dissolution <6.10> (1) L-Ascorbic acid—When the test is
Ascorbic Acid and Calcium performed at 50 revolutions per minute according to the
Paddle method, using 900 mL of water as the dissolution
Pantothenate Tablets medium, the dissolution rate in 60 minutes of Ascorbic Acid
and Calcium Pantothenate Tablets is not less than 85z.
アスコルビン酸･パントテン酸カルシウム錠
Start the test with 1 tablet of Ascorbic Acid and Calcium
Pantothenate Tablets, withdraw 20 mL of the medium at the
Ascorbic Acid and Calcium Pantothenate Tablets specified minute after starting the test, and filter through a
contain not less than 95.0z and not more than membrane filter with a pore size not exceeding 0.45 mm.
105.0z of the labeled amount of L-ascorbic acid Discard the first 10 mL of the filtrate, pipet V mL of the
(C6H8O6: 176.12) and not less than 93.0z and not subsequent filtrate, add 1st fluid for dissolution test to make
more than 107.0z of the labeled amount of calcium exactly V? mL so that each mL contains about 11 mg of L-
pantothenate (C18H32CaN2O10: 476.53). ascorbic acid (C6H8O6), and use this solution as the sample
solution. Separately, weigh accurately about 22 mg of Ascor-
bic Acid RS, previously dried in a desiccator (silica gel) for
with Ascorbic Acid and Calcium Pantothenate.
24 hours, dissolve in water to make exactly 100 mL, and
Identification (1) To a quantity of powdered Ascorbic warm at 379 C for 1 hour. Pipet 5 mL of this solution, add
448 Ascorbic Acid and Calcium Pantothenate Tablets / Official Monographs JP XVII
1st fluid for dissolution test to make exactly 100 mL, and use and not more than 2.0, respectively.
this solution as the standard solution. Determine the absor- System repeatability: When the test is repeated 6 times
bances, AT and AS, at 243 nm of the sample solution and with 100 mL of the standard solution under the above operat-
standard solution as directed under Ultraviolet-visible Spec- ing conditions, the relative standard deviation of the peak
trophotometry <2.24> within 1 hour after withdrawing the area of pantothenic acid is not more than 2.0z.
medium, using 1st fluid for dissolution test as the blank.
Assay (1) L-Ascorbic acid—Weigh accurately the mass of
Dissolution rate (z) with respect to the labeled amount not less than 20 tablets of Ascorbic Acid and Calcium Pan-
of L-ascorbic acid (C6H8O6) tothenate Tablets, and powder. Weigh accurately a portion
＝ MS × AT/AS × V?/V × 1/C × 45 of the powder, equivalent to about 0.1 g of L-ascorbic acid
(C6H8O6), add 50 mL of a solution of metaphosphoric acid
MS: Amount (mg) of Ascorbic Acid RS taken
(1 in 50), stir thoroughly, and titrate <2.50> with 0.05 mol/L
C: Labeled amount (mg) of L-ascorbic acid (C6H8O6) in 1
iodine VS (indicator: 1 mL of starch TS).
tablet
Each mL of 0.05 mol/L iodine VS ＝ 8.806 mg of C6H8O6
(2) Calcium pantothenate—When the test is performed
at 50 revolutions per minute according to the Paddle (2) Calcium pantothenate—Weigh accurately the mass
method, using 900 mL of water as the dissolution medium, of not less than 20 tablets of Ascorbic Acid and Calcium
the dissolution rate in 90 minutes of Ascorbic Acid and Cal- Pantothenate Tablets, and powder. Weigh accurately a por-
cium Pantothenate Tablets is not less than 75z. tion of the powder, equivalent to about 3 mg of calcium pan-
Start the test with 1 tablet of Ascorbic Acid and Calcium tothenate (C18H32CaN2O10), add exactly 20 mL of the inter-
Pantothenate Tablets, withdraw 20 mL of the medium at the nal standard solution, shake for 15 minutes, filter through a
specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm, and
membrane filter with a pore size not exceeding 0.45 mm. use the filtrate as the sample solution. Separately, weigh ac-
Discard the first 10 mL of the filtrate, pipet V mL of the curately about 30 mg of Calcium Pantothenate RS (separate-
subsequent filtrate, add 1st fluid for dissolution test to make ly determine the loss on drying <2.41> under the same condi-
exactly V? mL so that each mL contains about 3.3 mg of cal- tions as Calcium Pantothenate), and dissolve in water to
cium pantothenate (C18H32CaN2O10), and use this solution as make exactly 50 mL. Pipet 5 mL of this solution, add exactly
the sample solution. Separately, weigh accurately about 16.5 20 mL of the internal standard solution, and use this solu-
mg of Calcium Pantothenate RS (separately determine the tion as the standard solution. Perform the test with 5 mL
loss on drying <2.41> under the same conditions as Calcium each of the sample solution and standard solution as directed
Pantothenate), and dissolve in water to make exactly 100 under Liquid Chromatography <2.01> according to the fol-
mL. Pipet 2 mL of this solution, add water to make exactly lowing conditions, and calculate the ratios, QT and QS, of
100 mL, and use this solution as the standard solution. Per- the peak area of pantothenic acid to that of the internal
form the test with exactly 100 mL each of the sample solution standard.
Amount (mg) of calcium pantothenate (C18H32CaN2O10)
＝ MS × QT/QS × 1/10
termine the peak areas, AT and AS, of pantothenic acid in
each solution. MS: Amount (mg) of Calcium Pantothenate RS taken, cal-
of calcium pantothenate (C18H32CaN2O10) Internal standard solution—A solution of acetaminophen
＝ MS × AT/AS × V?/V × 1/C × 18 (1 in 50,000).
MS: Amount (mg) of Calcium Pantothenate RS taken, cal-
length: 200 nm).
C: Labeled amount (mg) of calcium pantothenate
(C18H32CaN2O10) in 1 tablet
Operating conditions— gel for liquid chromatography (3 mm in particle diameter).
Detector: An ultraviolet absorption photometer (wave- Column temperature: A constant temperature of about
length: 210 nm). 259C.
Column: A stainless steel column 4.6 mm in inside diame- Mobile phase: A mixture of 0.05 mol/L sodium dihydro-
ter and 15 cm in length, packed with octadecylsilanized sili- gen phosphate TS and acetonitrile for liquid chromatogra-
cone polymer coated silica gel for liquid chromatography (5 phy (97:3).
mm in particle diameter). Flow rate: Adjust so that the retention time of pantothenic
Column temperature: A constant temperature of about acid is about 3 minutes.
359 C. System suitability—
Mobile phase: Dissolve 7.80 g of sodium dihydrogen phos- System performance: When the procedure is run with 5 mL
phate dihydrate in 900 mL of water, adjust to pH 2.6 with of the standard solution under the above operating condi-
phosphoric acid, and add water to make 1000 mL. To 970 tions, pantothenic acid and the internal standard are eluted
mL of this solution add 30 mL of acetonitrile for liquid in this order with the resolution between these peaks being
chromatography. not less than 4.
Flow rate: Adjust so that the retention time of pantothenic System repeatability: When the test is repeated 6 times
acid is about 10 minutes. with 5 mL of the standard solution under the above operating
System suitability— conditions, the relative standard deviation of the ratio of the
System performance: When the procedure is run with 100 peak area of pantothenic acid to that of the internal standard
mL of the standard solution under the above operating con- is not more than 1.0z.
factor of the peak of pantothenic acid are not less than 5000
JP XVII Official Monographs / Aspirin 449

L-Aspartic Acid phy <2.03>. Spot 5 mL each of the sample solution and stand-
ard solution on a plate of silica gel for thin-layer chromatog-
L-アスパラギン酸 raphy, develop with a mixture of 1-butanol, water and acetic
acid (100) (3:1:1) to a distance of about 10 cm, and dry the
plate at 809C for 30 minutes. Spray evenly a solution of nin-
hydrin in a mixture of methanol and acetic acid (100) (97:3)
(1 in 100), and heat at 809 C for 10 minutes: the spot other
C4H7NO4: 133.10 than the principal spot obtained with the sample solution is
(2S )-2-Aminobutanedioic acid not more intense than the spot obtained with the standard
[56-84-8] solution.
L-Aspartic
Acid, when dried, contains not less than
3 hours).
98.5z and not more than 101.0z of L-aspartic acid
(C4H7NO4). Residue on ignition <2.44> Not more than 0.1z (1 g).
Description L-Aspartic Acid occurs as white, crystals or Assay Weigh accurately about 0.15 g of L-Aspartic Acid,
crystalline powder. previously dried, dissolve in 50 mL of water by warming.
It is sparingly soluble in water, and practically insoluble in After cooling, titrate <2.50> with 0.1 mo/L sodium hydrox-
ethanol (99.5). ide VS (potentiometric titration). Perform a blank determi-
It dissolves in dilute hydrochloric acid and in 0.2 mol/L nation in the same manner, and make any necessary correc-
sodium hydroxide TS. tion.
Identification Determine the infrared absorption spectrum Each mL of 0.1 mol/L sodium hydroxide VS
of L-Aspartic Acid as directed in the potassium bromide disk ＝ 13.31 mg of C4H7NO4
numbers.
Aspirin
D : ＋24.0 – ＋26.09 (2 g, after
drying, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). Acetylsalicylic Acid
pH <2.54> Dissolve 0.4 g of L-Aspartic Acid in 100 mL of
アスピリン
water by warming, and allow to cool: between 2.5 and 3.5.
of L-Aspartic Acid in 20 mL of 1 mol/L hydrochloric acid
TS: the solution is clear and colorless.
(2) Chloride <1.03>—Dissolve 0.5 g of L-Aspartic Acid in
C9H8O4: 180.16
6 mL of dilute nitric acid and 20 mL of water, add water to
2-Acetoxybenzoic acid
make 50 mL, and perform the test with this solution as the
[50-78-2]
test solution. Prepare the control solution with 0.30 mL of
Aspirin, when dried, contains not less than 99.5z
(3) Sulfate <1.14>—Dissolve 0.6 g of L-Aspartic Acid in
of aspirin (C9H8O4).
5 mL of dilute hydrochloric acid and 30 mL of water, add
water to make 45 mL, and add 5 mL of barium chloride TS. Description Aspirin occurs as white crystals, granules or
Perform the test with this solution as the test solution. Pre- powder. It is odorless, and has a slight acid taste.
pare the control solution with 0.35 mL of 0.005 mol/L sulfu- It is freely soluble in ethanol (95) and in acetone, soluble
ric acid VS, add 5 mL of dilute hydrochloric acid and water in diethyl ether, and slightly soluble in water.
to make 45 mL, and add 5 mL of barium chloride (not more It dissolves in sodium hydroxide TS and in sodium car-
than 0.028z). bonate TS.
(4) Ammonium <1.02>—Perform the test with 0.25 g of In moist air, it gradually hydrolyzes to salicylic acid and
L-Aspartic Acid. Prepare the control solution with 5.0 mL of acetic acid.
Standard Ammonium Solution (not more than 0.02z). Melting point: about 1369C (bath fluid is heated at 1309 C
(5) Heavy metals <1.07>—Proceed with 1.0 g of L- previously).
Aspartic Acid according to Method 4, and perform the test.
Identification (1) Boil 0.1 g of Aspirin in 5 mL of water
for 5 to 6 minutes, cool, and add 1 to 2 drops of iron (III)
chloride TS: a red-purple color is produced.
(6) Iron <1.10>—Prepare the test solution with 1.0 g of L-
(2) Boil 0.5 g of Aspirin in 10 mL of sodium carbonate
Aspartic Acid according to Method 1, and perform the test
TS for 5 minutes, and add 10 mL of dilute sulfuric acid: the
odor of acetic acid is perceptible, and a white precipitate is
produced. Filter the precipitate, add 3 mL of ethanol (95)
(7) Related substances—Dissolve 0.20 g of L-Aspartic
and 3 mL of sulfuric acid to the filtrate, and heat: the odor
Acid in 10 mL of 0.2 mol/L sodium hydroxide TS, and use
of ethyl acetate is perceptible.
ple solution, add water to make exactly 10 mL. Pipet 1 mL Purity (1) Clarity of solution—Dissolve 0.5 g of Aspirin
of this solution, add water to make exactly 50 mL, and use in 10 mL of warm sodium carbonate TS: the solution is
this solution as the standard solution. Perform the test with clear.
450 Aspirin Tablets / Official Monographs JP XVII
(2) Salicylic acid—Dissolve 2.5 g of Aspirin in 25 mL of Identification (1) Weigh a quantity of powdered Aspirin
ethanol (95), and add 1.0 mL of this solution to a solution Tablets, equivalent to 0.1 g of Aspirin, add 10 mL of water,
which is prepared by transferring 1 mL of a freshly prepared and boil for 5 to 6 minutes. After cooling, filter, and add 1
dilute ammonium iron (III) sulfate TS to a Nessler tube and to 2 drops of iron (III) chloride TS to the filtrate: a red-violet
diluting with water to 50 mL. Allow to stand for 30 seconds: color develops.
the solution has no more color than the following control so- (2) Weigh a portion of powdered Aspirin Tablets,
lution. equivalent to 0.5 g of Aspirin, extract with two 10-mL por-
Control solution: Dissolve 0.100 g of salicylic acid in tions of warm ethanol (95), and filter the combined extracts.
water, and add 1 mL of acetic acid (100) and water to make Evaporate the filtrate to dryness, and boil the residue with 10
1000 mL. Add 1.0 mL of this solution to a solution which is mL of sodium carbonate TS for 5 minutes. Proceed as di-
prepared by transferring 1 mL of freshly prepared dilute am- rected in the Identification (2) under Aspirin.
monium iron (III) sulfate TS and 1 mL of ethanol (95) to a
Purity Salicylic acid—Take a portion of the powdered
Nessler tube and diluting with water to 50 mL. Allow to
Aspirin Tablets, equivalent to 1.0 g of Aspirin, shake with 15
stand for 30 seconds.
mL of ethanol (95) for 5 minutes, filter, discard the first 5
(3) Chloride <1.03>—Boil 1.8 g of Aspirin in 75 mL of
mL of the filtrate, and add 1.0 mL of the subsequent filtrate
water for 5 minutes, cool, add water to make 75 mL, and
to a solution which is prepared by transferring 1 mL of
filter. To 25 mL of the filtrate add 6 mL of dilute nitric acid
freshly prepared dilute ammonium iron (III) sulfate TS to a
and water to make 50 mL, and perform the test using this so-
Nessler tube and diluting with water to make 50 mL. Pro-
lution as the test solution. Prepare the control solution with
ceed as directed in the Purity (2) under Aspirin.
0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than
0.015z). Assay Weigh accurately and powder not less than 20
(4) Sulfate <1.14>—To 25 mL of the filtrate obtained in Aspirin Tablets. Weigh accurately a portion of the powder,
(3) add 1 mL of dilute hydrochloric acid and water to make equivalent to about 1.5 g of aspirin (C9H8O4), add exactly
50 mL. Perform the test using this solution as the test solu- 50 mL of 0.5 mol/L sodium hydroxide VS, and proceed as
tion. Prepare the control solution with 0.50 mL of 0.005 directed in the Assay under Aspirin.
(5) Heavy metals <1.07>—Dissolve 2.5 g of Aspirin in 30
＝ 45.04 mg of C9H8O4
mL of acetone, add 2 mL of dilute acetic acid and water to
make 50 mL, and perform the test using this solution as the Containers and storage Containers—Well-closed contain-
test solution. Prepare the control solution with 2.5 mL of ers.
Standard Lead Solution, 30 mL of acetone, 2 mL of dilute
acetic acid and water to make 50 mL (not more than 10
ppm). Aspirin Aluminum
(6) Readily carbonizable substances <1.15>—Weigh 0.5 g
of Aspirin, and perform the test. The solution has no more Aluminum Acetylsalicylate
color than Matching Fluid Q.
アスピリンアルミニウム
5 hours).
Assay Weigh accurately about 1.5 g of Aspirin, previously
dried, add exactly 50 mL of 0.5 mol/L sodium hydroxide
C18H15AlO9: 402.29
VS, and boil gently for 10 minutes under a reflux condenser
Bis(2-acetoxybenzoato)hydroxoaluminium
with a carbon dioxide-absorbing tube (soda lime). Cool, and
[23413-80-1]
titrate <2.50> immediately the excess sodium hydroxide with
0.25 mol/L sulfuric acid VS (indicator: 3 drops of phenol-
Aspirin Aluminum contains not less than 83.0z and
phthalein TS). Perform a blank determination.
not more than 90.0z of aspirin (C9H8O4: 180.16), and
Each mL of 0.5 mol/L sodium hydroxide VS not less than 6.0z and not more than 7.0z of alumi-
＝ 45.04 mg of C9H8O4 num (Al: 26.98), calculated on the anhydrous basis.
Containers and storage Containers—Well-closed contain- Description Aspirin Aluminum occurs as a white crystal-
ers. line powder. It is odorless or has a slight, acetic odor.
It is practically insoluble in water, in methanol, in ethanol
(95) and in diethyl ether.
Aspirin Tablets It dissolves, with decomposition, in sodium hydroxide TS
and in sodium carbonate TS.
Acetylsalicylic Acid Tablets Identification (1) Dissolve 0.1 g of Aspirin Aluminum in
10 mL of sodium hydroxide TS by heating, if necessary.
アスピリン錠
Neutralize 2 mL of the solution with hydrochloric acid, and
add 1 to 2 drops of iron (III) chloride TS: a red-purple color
Aspirin Tablets contain not less than 95.0z and not develops.
more than 105.0z of the labeled amount of aspirin (2) Determine the absorption spectrum of the sample so-
(C9H8O4: 180.16). lution obtained in the Assay (1) as directed under Ultravio-
let-visible Spectrophotometry <2.24>: it exhibits a maximum
with Aspirin.
(3) Place 2 g of Aspirin Aluminum in a platinum cruci-
JP XVII Official Monographs / Aspoxicillin Hydrate 451
ble, and ignite until charred. To the residue add 1 g of anhy- absorbance AS3 of the standard solution (2) at 278 nm.
drous sodium carbonate, and ignite for 20 minutes. After
Amount (mg) of aspirin (C9H8O4)
cooling, to the residue add 15 mL of dilute hydrochloric
acid, shake, and filter: the filtrate responds to the Qualita-  AT2 × AS1 
AT1 －
tive Tests <1.09> for aluminum salt.
＝ MS ×  AS2

Purity (1) Salicylate—Using AT2 and AS2 obtained in the  AS3 
Assay (1), calculate the amount of salicylate [as salicylic acid
MS: Amount (mg) of Aspirin RS taken
(C7H6O3: 138.12)] by the following equation: salicylate con-
tent is not more than 7.5z, calculated on the anhydrous (2) Aluminum—Weigh accurately about 0.4 g of Aspirin
basis. Aluminum, and dissolve in 10 mL of sodium hydroxide TS.
Add dropwise 1 mol/L hydrochloric acid TS to adjust the
Amount (mg) of salicylic acid (C7H6O3)
solution to a pH of about 1, add 20 mL of acetic acid-
＝ MS × AT2/AS2 × 1/4
ammonium acetate buffer solution (pH 3.0) and 0.5 mL of
MS: Amount (mg) of salicylic acid for assay taken Cu-PAN TS, and heat. While boiling, titrate <2.50> with
0.05 mol/L disodium dihydrogen ethylenediamine tetraace-
(2) Heavy metals <1.07>—Place 2.0 g of Aspirin Alumi-
tate VS until the color of the solution changes from red to
num in a porcelain crucible, cover the crucible loosely, and
yellow and persists for 1 minute. Perform a blank determi-
ignite at a low temperature until charred. After cooling, add
nation, and make any necessary correction.
2 mL of nitric acid and 1 mL of sulfuric acid to the content
of the crucible, heat gently the crucible until white fumes are Each mL of 0.05 mol/L disodium dihydrogen
evolved, and continue the heating until white fumes are no ethylenediamine tetraacetate VS
longer evolved, then ignite between 5009 C and 6009C until ＝ 1.349 mg of Al
the carbon is incinerated. When the incineration is not com-
pleted, add 2 mL of nitric acid and 1 mL of sulfuric acid,
ers.
and heat gently in the same manner, then ignite between
5009C and 6009C to incinerate completely. After cooling,
add 2 mL of hydrochloric acid, and proceed as directed in
Method 2, and perform the test. Prepare the control solution Aspoxicillin Hydrate
by using the same quantities of the same reagents as directed
アスポキシシリン水和物
for the preparation of the test solution, and add 2.0 mL of
Standard Lead Solution and water to make 50 mL (not more
than 10 ppm).
(3) Arsenic <1.11>—Dissolve 1.0 g of Aspirin Aluminum
in 15 mL of sodium hydroxide TS. To this solution add 1
drop of phenolphthalein TS, and with stirring, add dropwise
hydrochloric acid until the red color of the solution disap-
pears. Then add 2 mL of hydrochloric acid, cool with occa-
sional shaking for 10 minutes, and filter with a glass filter C21H27N5O7S.3H2O: 547.58
(G3). Wash the residue with two 5 mL portions of 1 mol/L (2S,5R,6R)-6-[(2R)-2-[(2R)-2-Amino-3-
hydrochloric acid TS, and combine the filtrate and the wash- methylcarbamoylpropanoylamino]-
ings. Use this solution as the test solution, and perform the 2-(4-hydroxyphenyl)acetylamino]-3,3-dimethyl-7-oxo-
test (not more than 2 ppm). 4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid
trihydrate
Water <2.48> Not more than 4.0z (0.15 g, direct titration).
[63358-49-6, anhydride]
Assay (1) Aspirin—Weigh accurately about 0.1 g of Aspi-
rin Aluminum, add 40 mL of sodium fluoride TS, and shake Aspoxicillin Hydrate contains not less than 950 mg
for 5 minutes. Allow the solution to stand for 10 minutes (potency) and not more than 1020 mg (potency) per
with frequent shaking. Extract the solution with six 20-mL mg, calculated on the anhydrous basis. The potency of
portions of chloroform. Combine all chloroform extracts, Aspoxicillin Hydrate is expressed as mass (potency) of
and add chloroform to make exactly 200 mL. Measure ex- aspoxicillin (C21H27N5O7S: 493.53).
actly 10 mL of this solution, add chloroform to make exactly
Description Aspoxicillin Hydrate occurs as a white, crys-
tals or crystalline powder.
rately, weigh accurately about 90 mg of salicylic acid for
It is freely soluble in N, N-dimethylformamide, sparingly
assay, previously dried in a desiccator (silica gel) for 3 hours,
soluble in water, and practically insoluble in acetonitrile, in
and dissolve in chloroform to make exactly 200 mL. Meas-
methanol and in ethanol (95).
ure exactly 5 mL of this solution, add chloroform to make
exactly 200 mL, and use this solution as the standard solu- Identification (1) Determine the absorption spectrum of a
tion (1). Then weigh accurately about 90 mg of Aspirin RS, solution of Aspoxicillin Hydrate (1 in 4000) as directed
previously dried in a desiccator (silica gel) for 5 hours, and under Ultraviolet-visible Spectrophotometry <2.24>, and
dissolve in chloroform to make exactly 200 mL. Measure ex- compare the spectrum with the Reference Spectrum or the
actly 10 mL of this solution, add chloroform to make exactly spectrum of a solution of Aspoxicillin RS prepared in the
100 mL, and use this solution as the standard solution (2). same manner as the sample solution: both spectra exhibit
Perform the test with these solutions as directed under Ultra- similar intensities of absorption at the same wavelengths.
violet-visible Spectrophotometry <2.24>. Determine the ab- (2) Determine the infrared absorption spectrum of
sorbances, AT1 and AS1, of the sample solution and standard Aspoxicillin Hydrate as directed in the potassium bromide
solution (1) at 278 nm, and absorbances, AT2 and AS2, of disk method under Infrared Spectrophotometry <2.25>, and
these solution, at 308 nm, respectively. Then determine the
452 Atenolol / Official Monographs JP XVII
compare the spectrum with the Reference Spectrum or spec- MS: Amount [mg (potency)] of Aspoxicillin RS taken
trum of Aspoxicillin RS: both spectra exhibit similar intensi-
Internal standard solution—A solution of N-(3-hydroxy-
phenyl)acetamide (1 in 1000).
D : ＋170 – ＋1859(0.2 g calcu- Operating conditions—
lated on the anhydrous bases, water, 20 mL, 100 mm). Detector: An ultraviolet absorption photometer (wave-
length: 280 nm).
pH <2.54> Dissolve 1.0 g of Aspoxicillin Hydrate in 50 mL
of water: the pH of the solution is between 4.2 and 5.2.
Purity (1) Clarity and color of solution—Dissolve 1.0 g gel for liquid chromatography (5 mm in particle diameter).
of Aspoxicillin Hydrate in 50 mL of water: the solution is Column temperature: A constant temperature of about
clear and colorless. 409C.
(2) Heavy metals <1.07>—Proceed with 2.0 g of Aspoxi- Mobile phase: To 130 mL of acetonitrile add potassium
cillin Hydrate according to Method 2, and perform the test. dihydrogenphosphate TS (pH 3.0) to make 1000 mL.
Prepare the control solution with 2.0 mL of Standard Lead Flow rate: Adjust so that the retention time of aspoxicillin
Solution (not more than 10 ppm). is about 3 minutes.
(3) Arsenic <1.11>—Prepare the test solution with 2.0 g System suitability—
of Aspoxicillin Hydrate according to Method 5, and perform System performance: When the procedure is run with 10
the test (not more than 1 ppm). mL of the standard solution under the above operating con-
(4) Related substances—Dissolve 0.05 g of Aspoxicillin ditions, aspoxicillin and the internal standard are eluted in
Hydrate in 10 mL of the mobile phase, and use this solution this order with the resolution between these peaks being not
as the sample solution. Pipet 1 mL of the sample solution, less than 8.
add the mobile phase to make exactly 100 mL, and use this System repeatability: When the test is repeated 6 times
solution as the standard solution. Perform the test with with 10 mL of the standard solution under the above operat-
exactly 10 mL each of these solutions as directed under ing conditions, the relative standard deviation of the ratios
Liquid Chromatography <2.01> according to the following of the peak area of aspoxicillin to that of the internal stand-
conditions, and determine the areas of each peak by the au- ard is not more than 0.8z.
tomatic integration method: the area of each peak other than
aspoxicillin from the sample solution is not larger than 3/10
times the peak area of aspoxicillin from the standard solu-
tion, and the total of peak areas other than aspoxicillin from
the sample solution is not larger than the peak area of Atenolol
aspoxicillin from the standard solution.
アテノロール
the Assay.
retention time of aspoxicillin.
C14H22N2O3: 266.34
2-(4-{(2RS )-2-Hydroxy-3-
[(1-methylethyl)amino]propyloxy}phenyl)acetamide
[29122-68-7]
solution, add the mobile phase to make exactly 10 mL. Con-
Atenolol, when dried, contains not less than 99.0z
firm that the peak area of aspoxicillin obtained from 10 mL
and not more than 101.0z of atenolol (C14H22N2O3).
of this solution is equivalent to 15 to 25z of that of aspoxi-
cillin obtained from 10 mL of the standard solution. Description Atenolol occurs as a white to pale yellow crys-
System repeatability: When the test is repeated 6 times talline powder.
with 10 mL of the standard solution under the above operat- It is freely soluble in methanol and in acetic acid (100),
ing conditions, the relative standard deviation of the peak soluble in ethanol (99.5), and slightly soluble in water.
areas of aspoxicillin is not more than 5z. A solution of Atenolol in methanol (1 in 25) shows no op-
tical rotation.
Water <2.48> Not less than 9.5z and not more than 13.0z
(0.2 g, volumetric titration, direct titration). Identification (1) Determine the absorption spectrum of a
solution of Atenolol in methanol (1 in 50,000) as directed
Assay Weigh accurately an amount of Aspoxicillin Hy-
under Ultraviolet-visible Spectrophotometry <2.24>, and
drate and Aspoxicillin RS, equivalent to about 0.1 g (po-
compare the spectrum with the Reference Spectrum: both
tency), dissolve each in a suitable amount of water, add
exactly 10 mL of the internal standard solution, 6.5 mL of
wavelengths.
acetonitrile and water to make 50 mL, and use these solu-
tions as the sample solution and the standard solution,
Atenolol as directed in the potassium bromide disk method
respectively. Perform the test with 10 mL each of these solu-
tions as directed under Liquid Chromatography <2.01> ac-
QT and QS, of the peak area of aspoxicillin to that of the in-
ternal standard. Melting Pint <2.60> 152 – 1569C
Amount [ mg (potency)] of aspoxicillin (C21H27N5O7S) Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
＝ MS × QT/QS × 1000 Atenolol according to Method 2, and perform the test. Pre-
JP XVII Official Monographs / Atorvastatin Calcium Hydrate 453

lution (not more than 20 ppm). Atorvastatin Calcium Hydrate
(2) Related substances—Dissolve 50 mg of Atenolol in
25 mL of the mobile phase, and use this solution as the sam- アトルバスタチンカルシウム水和物
ple solution. Pipet 1 mL of the sample solution, add the mo-
bile phase to make exactly 200 mL, and use this solution as
lowing conditions, and determine each peak area by the
automatic integration method: the area of the peak other
than atenolol obtained with the sample solution is not larger
than 1/2 times the peak area of atenolol obtained with the
standard solution, and the total area of the peaks other than
atenolol with the sample solution is not larger than the peak
C66H68CaF2N4O10.3H2O: 1209.39
area of atenolol with the standard solution.
Monocalcium bis{(3R,5R )-7-[2-(4-fluorophenyl)-
5-(1-methylethyl)-3-phenyl-4-(phenylcarbamoyl)-1H-pyrrol-1-yl]-
3,5-dihydroxyheptanoate} trihydrate
length: 226 nm).
[344423-98-9]
Atorvastatin Calcium Hydrate contains not less
than 98.0z and not more than 102.0z of atorvastatin
calcium (C66H68CaF2N4O10: 1155.34), calculated on the
259 C.
anhydrous basis.
phosphate in 1000 mL of water, and adjust to pH 3.0 with Description Atorvastatin Calcium Hydrate occurs as a
phosphoric acid. To 40 volume of this solution add 9 volume white to pale yellowish white crystalline powder.
of methanol and 1 volume of tetrahydrofuran. Dissolve 1 g It is very soluble in methanol, freely soluble in dimethyl-
of sodium 1-octanesulfonate and 0.4 g of tetrabutylammo- sulfoxide, and very slightly soluble in water and in ethanol
nium hydrogensulfate in 1000 mL of this solution. (99.5).
Flow rate: Adjust so that the retention time of atenolol is It gradually turns yellowish white on exposure to light.
about 8 minutes. It shows crystal polymorphism.
retention time of atenolol.
solution of Atorvastatin Calcium Hydrate in methanol (1 in
ence Spectrum or the spectrum of a solution of Atorvastatin
mL. Confirm that the peak area of atenolol obtained with 10
Calcium RS prepared in the same manner as the sample solu-
mL of this solution is equivalent to 14 to 26z of that ob-
tion: both spectra exhibit similar intensities of absorption at
the same wavelengths.
(2) Determine the infrared absorption spectrum of Ator-
vastatin Calcium Hydrate as directed in the potassium bro-
factor of the peak of atenolol are not less than 5000 and not
more than 1.5, respectively.
the spectrum of Atorvastatin Calcium RS: both spectra
numbers. If any difference appears between the spectra,
recrystallize the sample and the reference standard according
area of atenolol is not more than 1.0z.
to the method otherwise specified, filter and dry the crystals,
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, and perform the test with the crystals.
3 hours). (3) A gruel-like liquid of Atorvastatin Calcium Hydrate
prepared by adding a small amount of dilute hydrochloric
acid responds to the Qualitative Tests <1.09> (1) for calcium
Assay Weigh accurately about 0.3 g of Atenolol, previ- salt. A solution of Atorvastatin Calcium Hydrate in a mix-
ously dried, dissolve in 100 mL of acetic acid (100), and ture of methanol and water (7:3) (1 in 250) is also responds
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- to the Qualitative Tests <1.09> (3) for calcium salt.
D : －7 – －109(0.2 g, calculated
on the anhydrous basis, dimethylsulfoxide, 20 mL, 100 mm).
＝ 26.63 mg of C14H22N2O3
Atorvastatin Calcium Hydrate according to Method 4, and
Containers and storage Containers—Tight containers. perform the test. Prepare the control solution with 1.0 mL of
(2) Related substances—Dissolve 20 mg of Atorvastatin
Calcium Hydrate in 20 mL of a mixture of water and aceto-
nitrile (1:1), and use this solution as the sample solution.
454 Atorvastatin Calcium Tablets / Official Monographs JP XVII
Pipet 1 mL of the sample solution, add a mixture of water determine the water <2.48> in the same manner as Atorvasta-
and acetonitrile (1:1) to make exactly 100 mL, and use this tin Calcium Hydrate), dissolve each in an adequate amount
solution as the standard solution. Perform the test with ex- of a mixture of water and acetonitrile (1:1), add exactly 10
actly 20 mL each of the sample solution and standard solu- mL of the internal standard solution, then add a mixture of
tion as directed under Liquid Chromatography <2.01> ac- water and acetonitrile (1:1) to make 50 mL, and use these so-
cording to the following conditions, and determine each lutions as the sample solution and the standard solution, re-
peak area by the automatic integration method: the area of spectively. Perform the test with 10 mL each of the sample
the peak, having the relative retention time of about 0.8 to solution and standard solution as directed under Liquid
atorvastatin, obtained from the sample solution is not larger Chromatography <2.01> according to the following condi-
than 3/10 times the peak area of atorvastatin obtained from tions, and calculate the ratios, QT and QS, of the peak area
the standard solution, the area of the peak other than ator- of atorvastatin to that of the internal standard.
vastatin and the peak mentioned above from the sample so-
Amount (mg) of atorvastatin calcium (C66H68CaF2N4O10)
lution is not larger than 1/10 times the peak area of ator-
＝ MS × QT/QS
vastatin from the standard solution, and the total area of the
peaks other than atorvastatin from the sample solution is not MS: Amount (mg) of Atorvastatin Calcium RS taken, cal-
larger than the peak area of atorvastatin from the standard culated on the anhydrous basis
solution.
Internal standard solution—A solution of butyl para-
hydroxybenzoate in a mixture of water and acetonitrile (1:1)
(1 in 1500).
length: 254 nm).
length: 254 nm).
409 C.
Mobile phase A: Dissolve 10.5 g of citric acid monohy-
drate in 900 mL of water, adjust to pH 5.0 with ammonia so-
409C.
lution (28), and add water to make 1000 mL. To 400 mL of
Mobile phase: Dissolve 10.5 g of citric acid monohydrate
this solution add 100 mL of acetonitrile and 100 mL of tetra-
in 900 mL of water, adjust to pH 4.0 with ammonia solution
hydrofuran.
(28), and add water to make 1000 mL. To 530 mL of this
Mobile phase B: A mixture of acetonitrile and tetra-
solution add 270 mL of acetonitrile and 200 mL of tetra-
hydrofuran (1:1).
hydrofuran.
Flow rate: Adjust so that the retention time of atorvasta-
tin is about 10 minutes.
Time after injection Mobile phase A Mobile phase B System performance: When the procedure is run with 10
of sample (min) (volz) (volz) mL of the standard solution under the above operating con-
ditions, the internal standard and atorvastatin are eluted in
0 – 40 93 7
40 – 80 93 → 60 7 → 40
less than 5.
Flow rate: Adjust so that the retention time of atorvasta- with 10 mL of the standard solution under the above operat-
tin is about 16 minutes. ing conditions, the relative standard deviation of the ratio of
Time span of measurement: About 5 times as long as the the peak area of atorvastatin to that of the internal standard
retention time of atorvastatin, beginning after the solvent is not more than 1.0z.
peak.
System suitability— Containers and storage Containers—Well-closed contain-
Test for required detectability: Pipet 5 mL of the standard ers.
solution, and add a mixture of water and acetonitrile (1:1) to Storage—Light-resistant.
make exactly 100 mL. Confirm that the peak area of ator-
vastatin obtained with 20 mL of this solution is equivalent to
3.5 to 6.5z of that obtained with 20 mL of the standard so- Atorvastatin Calcium Tablets
lution.
System performance: When the procedure is run with 20 アトルバスタチンカルシウム錠
ditions, the number of theoretical plates and the symmetry Atorvastatin Calcium Tablets contain not less
factor of the peak of atorvastatin are not less than 8000 and than 95.0z and not more than 105.0z of the
not more than 1.5, respectively. labeled amount of atorvastatin calcium hydrate
System repeatability: When the test is repeated 6 times (C66H68CaF2N4O10.3H2O: 1209.39).
ing conditions, the relative standard deviation of the peak Method of preparation Prepare as directed under Tablets,
area of atorvastatin is not more than 2.0z. with Atorvastatin Calcium Hydrate.
Water <2.48> 3.5 – 5.5z (50 mg, coulometric titration). Identification To a quantity of powdered Atorvastatin Cal-
cium Tablets, equivalent to 10 mg of Atorvastatin Calcium
Assay Weigh accurately about 20 mg each of Atorvastatin Hydrate, add 50 mL of methanol, shake thoroughly, and
Calcium Hydrate and Atorvastatin Calcium RS (separately centrifuge. To 2.5 mL of the supernatant liquid add metha-
JP XVII Official Monographs / Atorvastatin Calcium Tablets 455
nol to make 50 mL, and determine the absorption spectrum drate), and dissolve in a mixture of water and methanol (1:1)
of this solution as directed under Ultraviolet-visible Spectro- to make exactly 100 mL. Pipet 5 mL of this solution, add
photometry <2.24>: it exhibits a maximum between 244 nm water to make exactly 50 mL. Pipet 5 mL of this solution,
and 248 nm. add water to make exactly 50 mL, and use this solution as
under Liquid Chromatography <2.01>, and determine the
peak areas, AT and AS, of atorvastatin in each solution.
To 1 tablet of Atorvastatin Calcium Tablets add 3V/5 mL
of a mixture of water and methanol (1:1), and disintegrate Dissolution rate (z) with respect to the labeled amount
the tablet by shaking. Add exactly V/10 mL of the internal of atorvastatin calcium hydrate (C66H68CaF2N4O10.3H2O)
standard solution, and add a mixture of water and methanol ＝ MS × AT/AS × V?/V × 1/C × 9 × 1.047
(1:1) to make V mL so that each mL contains about 0.1 mg
MS: Amount (mg) of Atorvastatin Calcium RS taken, cal-
of atorvastatin calcium hydrate (C66H68CaF2N4O10.3H2O).
Centrifuge this solution, and use the supernatant liquid as
C: Labeled amount (mg) of atorvastatin calcium hydrate
the sample solution. Separately, weigh accurately about 22
(C66H68CaF2N4O10.3H2O) in 1 tablet
mg of Atorvastatin Calcium RS (separately determine the
water <2.48> in the same manner as Atorvastatin Calcium Operating conditions—
Hydrate), and dissolve in a mixture of water and methanol Proceed as directed in the operating conditions in the
(1:1) to make exactly 20 mL. Pipet 5 mL of this solution, Assay.
add exactly 5 mL of the internal standard solution, then add System suitability—
a mixture of water and methanol (1:1) to make 50 mL, and System performance: When the procedure is run with 50
use this solution as the standard solution. Perform the test mL of the standard solution under the above operating con-
with 10 mL each of the sample solution and standard solution ditions, the number of theoretical plates and the symmetry
as directed under Liquid Chromatography <2.01> according factor of the peak of atorvastatin are not less than 6000 and
to the following conditions, and calculate the ratios, QT and not more than 2.0, respectively.
QS, of the peak area of atorvastatin to that of the internal System repeatability: When the test is repeated 6 times
standard. with 50 mL of the standard solution under the above operat-
Amount (mg) of atorvastatin calcium hydrate
area of atorvastatin is not more than 2.0z.
(C66H68CaF2N4O10.3H2O)
＝ MS × QT/QS × V/200 × 1.047 Assay To 20 Atorvastatin Calcium Tablets add 3V/5 mL
of a mixture of water and methanol (1:1), and disintegrate
the tablet by shaking. Add exactly V/10 mL of the internal
standard solution, add a mixture of water and methanol
Internal standard solution—A solution of 1,3-dinitroben- (1:1) to make V mL so that each mL contains about 2 mg of
zene in methanol (1 in 2500). atorvastatin calcium hydrate (C66H68CaF2N4O10.3H2O), and
Operating conditions— centrifuge. To 2.5 mL of the supernatant liquid add a mix-
Proceed as directed in the operating conditions in the ture of water and methanol (1:1) to make 50 mL, and use
Assay. this solution as the sample solution. Separately, weigh accu-
System suitability— rately about 44 mg of Atorvastatin Calcium RS (separately
System performance: When the procedure is run with 10 determine the water <2.48> in the same manner as Atorvasta-
mL of the standard solution under the above operating con- tin Calcium Hydrate), add exactly 2 mL of the internal
ditions, the internal standard and atorvastatin are eluted in standard solution, and add a mixture of water and methanol
this order with the resolution between these peaks being not (1:1) to make 20 mL. Pipet 2.5 mL of this solution, add a
less than 10. mixture of water and methanol (1:1) to make 50 mL, and use
System repeatability: When the test is repeated 6 times this solution as the standard solution. Perform the test with
with 10 mL of the standard solution under the above operat- 10 mL each of the sample solution and standard solution as
ing conditions, the relative standard deviation of the ratio of directed under Liquid Chromatography <2.01> according to
the peak area of atorvastatin to that of the internal standard the following conditions, and calculate the ratios, QT and
is not more than 1.0z. QS, of the peak area of atorvastatin to that of the internal
standard.
tions per minute according to the Paddle method, using 900 Amount (mg) of atorvastatin calcium hydrate
mL of water as the dissolution medium, the dissolution rate (C66H68CaF2N4O10.3H2O)
in 15 minutes of Atorvastatin Calcium Tablets is not less in 1 tablet of Atorvastatin Calcium Tablets
than 80z. ＝ MS × QT/QS × V/400 × 1.047
Start the test with 1 tablet of Atorvastatin Calcium
membrane filter with a pore size not exceeding 0.45 mm. Dis- Internal standard solution—A solution of 1,3-dinitroben-
card the first 10 mL of the filtrate, pipet V mL of the subse- zene in methanol (1 in 125).
quent filtrate, add water to make exactly V? mL so that each Operating conditions—
mL contains about 6 mg of atorvastatin calcium hydrate Detector: An ultraviolet absorption photometer (wave-
(C66H68CaF2N4O10.3H2O), and use this solution as the sam- length: 244 nm).
ple solution. Separately, weigh accurately about 60 mg of Column: A stainless steel column 4.6 mm in inside diame-
Atorvastatin Calcium RS (separately determine the water ter and 25 cm in length, packed with octadecylsilanized silica
<2.48> in the same manner as Atorvastatin Calcium Hy- gel for liquid chromatography (5 mm in particle diameter).
456 Atropine Sulfate Hydrate / Official Monographs JP XVII
Column temperature: A constant temperature of about sponds to the Qualitative Tests <1.09> for sulfate.
309 C.
Purity (1) Clarity and color of solution —Dissolve 0.5 g
Mobile phase: Dissolve 10.5 g of citric acid monohydrate
of Atropine Sulfate Hydrate in 10 mL of water: the solution
in 900 mL of water, adjust to pH 4.0 with ammonia solution
is clear and colorless.
(2) Acidity—Dissolve 1.0 g of Atropine Sulfate Hydrate
solution add 270 mL of acetonitrile and 200 mL of tetra-
in 20 mL of water, and add 0.30 mL of 0.02 mol/L sodium
hydrofuran.
hydroxide VS and 1 drop of methyl red-methylene blue TS: a
Flow rate: Adjust so that the retention time of atorvasta-
green color develops.
tin is about 9 minutes.
(3) Related substances—Dissolve 0.25 g of Atropine Sul-
fate Hydrate in 1 mL of diluted hydrochloric acid (1 in 10),
add water to make 15 mL, and use this solution as the sam-
ple solution.
ditions, the internal standard and atorvastatin are eluted in
(i) To 5 mL of the sample solution add 2 to 3 drops of
hydrogen hexachloroplatinate (IV) TS: no precipitate is
less than 10.
formed.
(ii) To 5 mL of the sample solution add 2 mL of ammo-
nia TS, and shake vigorously: the turbidity of the solution is
not greater than that of the following control solution.
the peak area of atorvastatin to that of the internal standard
Control solution: To 0.30 mL of 0.01 mol/L hydrochloric
acid VS add 6 mL of dilute nitric acid and water to make 50
Containers and storage Containers—Tight containers. mL. To this solution add 1 mL of silver nitrate TS, and
allow 7 mL of the mixture to stand for 5 minutes.
(4) Hyoscyamine—Weigh accurately about 1 g of Atro-
Atropine Sulfate Hydrate pine Sulfate Hydrate, previously dried, and dissolve in water
to make exactly 10 mL: the specific optical rotation [a]20 D
アトロピン硫酸塩水和物 <2.49> of this solution in a 100-mm cell is between －0.609
and ＋0.109.
(5) Readily carbonizable substances <1.15>—Take 0.20 g
of Atropine Sulfate Hydrate, and perform the test: the solu-
tion has no more color than Matching Fluid A.
um, phosphorus (V) oxide, 1109C, 4 hours).
(C17H23NO3)2.H2SO4.H2O: 694.83 Residue on ignition <2.44> Not more than 0.1z (0.5 g).
(1R,3r,5S )-8-Methyl-8-azabicyclo[3.2.1]oct-3-yl [(2RS )-
Assay Dissolve about 0.25 g of Atropine Sulfate Hydrate,
3-hydroxy-2-phenyl]propanoate hemisulfate hemihydrate
previously dried and accurately weighed, in 30 mL of acetic
[5908-99-6]
acid (100). If necessary, dissolve it by warming, and cool.
Titrate <2.50> with 0.05 mol/L perchloric acid VS until the
Atropine Sulfate Hydrate, when dried, contains not
color of the solution changes from purple through blue to
less than 98.0z of atropine sulfate [(C17H23NO3)2.
blue-green (indicator: 3 drops of crystal violet TS). Perform
H2SO4: 676.82].
a blank determination, and make any necessary correction.
Description Atropine Sulfate Hydrate occurs as colorless
crystals or a white crystalline powder. It is odorless.
＝ 33.84 mg of (C17H23NO3)2.H2SO4
It is very soluble in water and in acetic acid (100), freely
soluble in ethanol (95), and practically insoluble in diethyl Containers and storage Containers—Tight containers.
ether. Storage—Light-resistant.
Melting point: 188 – 1949 C (with decomposition). Intro-
duce a capillary tube charged with dried sample into a bath
previously heated to 1809 C, and continue to heat at a rate of Atropine Sulfate Injection
rise of about 39C per minute.
It is affected by light. アトロピン硫酸塩注射液
Identification (1) To 1 mg of Atropine Sulfate Hydrate
add 3 drops of fuming nitric acid, and evaporate the mixture Atropine Sulfate Injection is an aqueous injection.
on a water bath to dryness. Dissolve the residue in 1 mL of It contains not less than 93.0z and not more than
N, N-dimethylformamide, and add 5 to 6 drops of tetraethyl- 107.0z of the labeled amount of atropine sulfate
ammonium hydroxide TS: a red-purple color develops. hydrate [(C17H23NO3)2.H2SO4.H2O: 694.83].
(2) To 2 mL of a solution of Atropine Sulfate Hydrate
(1 in 50) add 4 to 5 drops of hydrogen tetrachloroaurate (III)
tions, with Atropine Sulfate Hydrate.
TS: a lusterless, yellowish white precipitate is formed.
(3) To 5 mL of a solution of Atropine Sulfate Hydrate Description Atropine Sulfate Injection is a clear, colorless
(1 in 25) add 2 mL of ammonia TS, and allow to stand for 2 liquid.
to 3 minutes. Collect the precipitate, wash with water, and pH: 4.0 – 6.0
dry in a desiccator (in vacuum, silica gel) for 4 hours: it melts
Identification (1) Evaporate a volume of Atropine Sul-
<2.60> between 1159C and 1189C.
fate Injection, equivalent to 1 mg of Atropine Sulfate Hy-
(4) A solution of Atropine Sulfate Hydrate (1 in 20) re-
drate, on a water bath to dryness. Proceed with the residue
JP XVII Official Monographs / Auranofin 457
as directed in the Identification (1) under Atropine Sulfate adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL
Hydrate. of this solution add 70 mL of tetrahydrofuran, and mix.
(2) Evaporate an exactly measured volume of Atropine Flow rate: Adjust so that the retention time of atropine is
Sulfate Injection, equivalent to 5 mg of Atropine Sulfate about 16 minutes.
Hydrate, on a water bath to dryness. After cooling, dissolve System suitability—
the residue in 1 mL of ethanol (95), and use this solution as System performance: When the procedure is run with 20
the sample solution. If insoluble substance remains, crush it, mL of the standard solution under the above operating con-
allow to stand, and use the supernatant liquid as the sample ditions, the internal standard and atropine are eluted in this
solution. Separately, dissolve 10 mg of Atropine Sulfate RS order with the resolution between these peaks being not less
in 2 mL of ethanol (95), and use this solution as the standard than 3.
solution. Perform the test with these solutions as directed System repeatability: When the test is repeated 6 times
under Thin-layer Chromatography <2.03>. Spot 5 mL each of with 20 mL of the standard solution under the above operat-
the sample solution and standard solution on a plate of silica ing conditions, the relative standard deviation of the ratio of
gel for thin-layer chromatography. Develop the plate with a the peak area of atropine to that of the internal standard is
mixture of acetone, water and ammonia water (28) (90:7:3) not more than 1.5z.
to a distance of about 10 cm, and dry the plate at 809C for
10 minutes. After cooling, spray evenly Dragendorff's TS
for spraying on the plate: the spots obtained from the sample
solution and the standard solution show an orange color and
the same R f value.
(3) Atropine Sulfate Injection responds to the Qualita- Auranofin
tive Tests <1.09> for sulfate.
オーラノフィン
Bacterial endotoxins <4.01> Less than 75 EU/mg.
Extractable volume <6.05> It meets the requirements.
ment.
Sterility <4.06> Perform the test: it meets the requirement.
Assay To an exactly measured volume of Atropine Sulfate
C20H34AuO9PS: 678.48
Injection, equivalent to about 5 mg of atropine sulfate hy-
(2,3,4,6-Tetra-O-acetyl-1-thio-
drate [(C17H23NO3)2.H2SO4.H2O], add exactly 3 mL of the
b-D-glucopyranosato)(triethylphosphine)gold
internal standard solution and water to make 50 mL, and use
[34031-32-8]
this solution as the sample solution. Separately, weigh accu-
rately about 25 mg of Atropine Sulfate RS (separately deter-
Auranofin, when dried, contains not less than
mine the loss on drying <2.41> under the same conditions as
98.0z and not more than 102.0z of auranofin
Atropine Sulfate Hydrate), and dissolve in water to make ex-
(C20H34AuO9PS).
actly 50 mL. Pipet 10 mL of this solution, add exactly 3 mL
of the internal standard solution and water to make 50 mL, Description Auranofin occurs as a white crystalline pow-
and use this solution as the standard solution. Perform the der.
test with 20 mL each of the sample solution and standard so- It is very soluble in chloroform, freely soluble in metha-
lution as directed under Liquid Chromatography <2.01> ac- nol, sparingly soluble in ethanol (99.5), and practically in-
cording to the following conditions, and calculate the ratios, soluble in water.
QT and QS, of the peak area of atropine to that of the inter- It shows crystal polymorphism.
nal standard.
Identification (1) To 50 mg of Auranofin add 3 mL of
Amount (mg) of atropine sulfate hydrate water, 3 mL of nitric acid and 3 mL of sulfuric acid, shake,
[(C17H23NO3)2.H2SO4.H2O] and allow to stand: golden colored suspended matters are
＝ MS × QT/QS × 1/5 × 1.027 produced.
MS: Amount (mg) of Atropine Sulfate RS taken, calcu-
Auranofin as directed in the paste method under Infrared
lated based on the dried basis
Internal standard solution—A solution of etilefrine hydro- the Reference Spectrum or the spectrum of Auranofin RS:
chloride (1 in 1000). both spectra exhibit similar intensities of absorption at the
Operating conditions— same wave numbers.
Detector: An ultraviolet absorption photometer (wave- (3) Prepare the test solution with 1 mg of Auranofin as
length: 210 nm). directed under Oxygen Flask Combustion Method <1.06>,
Column: A stainless steel column 4.6 mm in inside diame- using 10 mL of water as the absorbing liquid. Wash out the
ter and 15 cm in length, packed with octadecylsilanized silica test solution into a Nessler tube with water to make 30 mL.
gel for liquid chromatography (5 mm in particle diameter). Add 10 mL of dilute sulfuric acid, 3 mL of hexaammonium
Column temperature: A constant temperature of about heptamolybdate-sulfuric acid TS and 0.1 mL of tin (II) chlo-
409 C. ride TS, shake, and allow to stand for 10 to 15 minutes: a
Mobile phase: To 0.4 g of sodium lauryl sulfate add 500 blue color is developed.
mL of diluted phosphoric acid (1 in 1000) to dissolve, and
D : －54.0 – －62.09(after dry-
458 Auranofin Tablets / Official Monographs JP XVII
ing, 0.2 g, methanol, 20 mL, 100 mm). lowing conditions, and calculate the ratios, QT and QS, of
the peak area of auranofin to that of the internal standard.
Amount (mg) of auranofin (C20H34AuO9PS)
Purity (1) Chloride <1.03>—Put 0.5 g of Auranofin in a
＝ M S × Q T / QS
porcelain crucible, add 0.25 g of anhydrous sodium carbon-
ate, mix well, and ignite until the carbonized substance is dis- MS: Amount (mg) of Auranofin RS taken
appeared. After cooling, add 20 mL of water, heat, and
filter after cooling. Wash the residue with 20 mL of water,
hydroxybenzoate in a mixture of water and acetonitrile (1:1)
combine the filtrate and the washings, neutralize with dilute
(3 in 1250).
nitric acid, then add 6 mL of dilute nitric acid and water to
solution. Prepare the control solution as follows: Dissolve
length: 230 nm).
0.25 g of anhydrous sodium carbonate in 20 mL of water,
neutralize with dilute nitric acid, add 0.50 mL of 0.01 mol/L
hydrochloric acid, 6 mL of dilute nitric acid, and water to
259C.
Auranofin according to Method 2, and perform the test.
Mobile phase: A mixture of sodium dihydrogen phosphate
dihydrate solution (1 in 100), tetrahydrofuran and aceto-
nitrile (12:5:3).
(3) Arsenic <1.11>—Put 0.5 g of Auranofin in a Kjeldahl
Flow rate: Adjust so that the retention time of auranofin
flask, add cautiously 2 mL of sulfuric acid and 5 mL of
is about 6 minutes.
nitric acid, and heat until the solution becomes almost color-
less. After cooling, add 15 mL of a saturated solution of am-
monium oxalate monohydrate, heat until white fumes are
evolved, and concentrate to 1 to 2 mL. Then, add 3 mL of
ditions, auranofin and the internal standard are eluted in this
water and 1 drop of methyl orange TS, neutralize with am-
monia solution (28), filter, and perform the test using the fil-
than 9.
trate as the test solution: the color is not darker than that of
the following control solution.
Control solution: Heat a mixture of 2 mL of sulfuric acid
and 5 mL of nitric acid until white fumes are no longer
the peak area of auranofin to that of the internal standard is
evolved. After cooling, add 15 mL of a saturated solution of
not more than 1.0z.
ammonium oxalate monohydrate, heat until white fumes are
evolved, and concentrate to 1 to 2 mL. Add 3 mL of water Containers and storage Containers—Tight containers.
and 1 drop of methyl orange TS, neutralize with ammonia
solution (28), and filter. To the filtrate add 2.0 mL of Stand-
ard Arsenic Solution, then proceed in the same manner as Auranofin Tablets
for the test solution (not more than 4 ppm).
(4) Related substances—Dissolve 50 mg of Auranofin in オーラノフィン錠
5 mL of chloroform, and use this solution as the sample so-
lution. Pipet 1 mL of the sample solution, and add chlo-
Auranofin Tablets contain not less than 93.0z
roform to make exactly 100 mL. To exactly 3 mL of this so-
lution add chloroform to make exactly 10 mL, and use this
auranofin (C20H34AuO9PS: 678.48).
solutions as directed under Thin-layer Chromatography Method of preparation Prepare as directed under Tablets,
<2.03>. Spot 10 mL each of the sample solution and standard with Auranofin.
Identification Put an amount of powdered Auranofin
phy. Develop the plate with a mixture of chloroform and
Tablets, equivalent to 11 mg of Auranofin, in a porcelain
acetone (4:1) to a distance of about 10 cm, and air-dry the
crucible, and heat weakly to carbonize. After cooling, add
plate. Dry, furthermore, at 809C for 30 minutes. After cool-
2 mL of nitric acid and 5 drops of sulfuric acid, heat cau-
ing, allow the plate to stand in a iodine vapor for 30 minutes:
tiously at first then incinerate by ignition. After cooling, add
the spots other than the principal spot obtained from the
4 mL of aqua regia to the residue, dissolve by warming, and
sample solution are not more intense than the spot obtained
add 16 mL of water. To 5 mL of this solution add 0.5 mL of
tin (II) chloride TS: a purple to red-brown color is deve-
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, loped.
3 hours).
Assay Weigh accurately about 20 mg each of Auranofin ing to the following method: it meets the requirement of the
and Auranofin RS, both previously dried, dissolve each in Content uniformity test.
10 mL of a mixture of water and acetonitrile (1:1), and add To 1 tablet of Auranofin Tablets add 2 mL of water, dis-
exactly 5 mL each of the internal standard solution. Then integrate the tablet with the aid of ultrasonic waves,
add a mixture of water and acetonitrile (1:1) to make 100 add exactly 2 mL of the internal standard solution for every
mL, and use these solutions as the sample solution and the 3 mg of auranofin (C20H34AuO9PS), and add 2 mL of a mix-
standard solution, respectively. Perform the test with 10 mL ture of water and acetonitrile (1:1). Shake for 15 minutes,
each of the sample solution and standard solution as directed then add a mixture of water and acetonitrile (1:1) to make
under Liquid Chromatography <2.01> according to the fol- V mL so that each mL contains 0.3 mg of auranofin
JP XVII Official Monographs / Azathioprine 459
(C20H34AuO9PS), centrifuge, and use the supernatant liquid this solution as the standard solution. Perform the test with
as the sample solution. Then, proceed as directed in the 10 mL each of the sample solution and standard solution as
Assay. directed under Liquid Chromatography <2.01> according to
QS, of the peak area of auranofin to that of the internal
＝ MS × QT/QS × V/100
standard.
MS: Amount (mg) of Auranofin RS taken
Internal standard solution—A solution of butyl para- ＝ M S × QT / QS × 2
hydroxybenzoate in acetonitrile (9 in 10,000).
in 15 minutes of Auranofin Tablets is not less than 85z.
Start the test with 1 tablet of Auranofin Tablets, withdraw
Assay under Auranofin.
ditions, auranofin and the internal standard are eluted in this
3.3 mg of auranofin (C20H34AuO9PS), and use this solution
than 9.
as the sample solution. Separately, weigh accurately about
30 mg of Auranofin RS, previously dried at 1059C for 3
hours, and dissolve in acetonitrile to make exactly 50 mL.
Pipet 5 mL of this solution, and add water to make exactly
the peak area of auranofin to that of the internal standard is
100 mL. Pipet 10 mL of this solution, add water to make ex-
not more than 1.0z.
Perform the test with exactly 50 mL each of the sample solu- Containers and storage Containers—Tight containers.
tion and standard solution as directed under Liquid Chroma-
tography <2.01> under the following conditions, and deter-
mine the peak areas, AT and AS, of auranofin in each solu- Azathioprine
tion.
アザチオプリン
of auranofin (C20H34AuO9PS)
＝ MS × AT/AS × V?/V × 1/C × 9
C: Labeled amount (mg) of auranofin (C20H34AuO9PS) in
1 tablet
Proceed as directed in the operating conditions in the C9H7N7O2S: 277.26
Assay under Auranofin. 6-(1-Methyl-4-nitro-1H-imidazol-5-ylthio)purine
System suitability— [446-86-6]
mL of the standard solution under the above operating con- Azathioprine, when dried, contains not less than
ditions, the number of theoretical plates and the symmetry 98.5z of azathioprine (C9H7N7O2S).
factor of the peak of auranofin are not less than 5000 and
Description Azathioprine is light yellow, crystals or crystal-
line powder. It is odorless.
It is sparingly soluble in N, N-dimethylformamide and in
pyridine, very slightly soluble in water and in ethanol (99.5),
and practically insoluble in chloroform and in diethyl ether.
area of auranofin is not more than 1.0z.
It dissolves in sodium hydroxide TS and in ammonia TS.
Assay Accurately weigh the mass of not less than 20 It is gradually colored by light.
Auranofin Tablets, and powder them. Weigh accurately a Melting point: about 2409C (with decomposition).
portion of the powder, equivalent to about 60 mg of
Identification (1) Dissolve 0.01 g of Azathioprine in 50
auranofin (C20H34AuO9PS), add 40 mL of water, disperse
mL of water by warming. To 5 mL of this solution add 1 mL
the particles with the aid of ultrasonic waves, then add ex-
of dilute hydrochloric acid and 0.01 g of zinc powder, and
actly 40 mL of the internal standard solution, add 40 mL of
allow to stand for 5 minutes: a yellow color is produced.
a mixture of water and acetonitrile (1:1), and shake for 15
Filter this solution: the filtrate responds to the Qualitative
minutes. To this solution add a mixture of water and aceto-
Tests <1.09> for primary aromatic amines, and a red color is
nitrile (1:1) to make 200 mL, centrifuge, and use the super-
produced.
natant liquid as the sample solution. Separately, weigh accu-
(2) Dissolve 0.01 g of Azathioprine in 50 mL of water by
rately about 30 mg of Auranofin RS, previously dried at
warming. To 1 mL of this solution add 0.5 mL of phos-
1059C for 3 hours, dissolve in 60 mL of a mixture of water
photungstic acid TS and 0.5 mL of dilute hydrochloric acid:
and acetonitrile (1:1), add exactly 20 mL of the internal
a white precipitate is formed.
standard solution, then add water to make 100 mL, and use
460 Azathioprine Tablets / Official Monographs JP XVII
(3) Prepare the test solution by proceeding with 0.03 g of Time span of measurement: About three times as long as
Azathioprine according to the Oxygen Flask Combustion the retention time of azathioprine, beginning after the sol-
Method <1.06>, using 20 mL of water as the absorbing liq- vent peak.
uid: the test solution responds to the Qualitative Tests <1.09> System suitability—
(1) for sulfate. Test for required detectability: To exactly 5 mL of the
(4) Dissolve 0.01 g of Azathioprine in 2 mol/L hydro- standard solution add water to make exactly 50 mL. Con-
chloric acid TS to make 100 mL. Dilute 5 mL of the solution firm that the peak area of azathioprine obtained from 20 mL
with water to make 50 mL. Determine the absorption spec- of this solution is equivalent to 8 to 12z of that of
trum of the solution as directed under Ultraviolet-visible azathioprine obtained from 20 mL of the standard solution.
Spectrophotometry <2.24>, and compare the spectrum with System performance: Dissolve 10 mg of Azathioprine in
the Reference Spectrum or the spectrum of a solution of 80 mL of water by warming, cool, and add water to make
Azathioprine RS prepared in the same manner as the sample 100 mL. To 2 mL of this solution add 2 mL of a solution,
solution: both spectra exhibit similar intensities of absorp- separately prepared by dissolving 0.06 g of benzoic acid in 3
tion at the same wavelengths. mL of methanol and diluting with water to make 10 mL, and
add the mobile phase to make 25 mL. When the procedure is
of Azathioprine in 50 mL of N, N-dimethylformamide: the
conditions, azathioprine and benzoic acid are eluted in this
solution is clear and shows a light yellow color.
(2) Acidity or alkalinity—Add 100 mL of water to 2.0 g
than 9.
of Azathioprine, shake well for 15 minutes, centrifuge for 5
minutes at 10,000 revolutions per minute, and filter. Discard
the first 20 mL of the filtrate, add 2 drops of methyl red TS
to 40 mL of the subsequent filtrate, and use this solution as
areas of azathioprine is not more than 2.0z.
the sample solution.
(i) Add 0.10 mL of 0.02 mol/L hydrochloric acid VS to Loss on drying <2.41> Not more than 0.5z (1 g, 1059C,
20 mL of the sample solution: a red color develops. 5 hours).
(ii) Add 0.10 mL of 0.02 mol/L sodium hydroxide VS to
20 mL of the sample solution: a yellow color develops.
(3) Sulfate <1.14>—To 25 mL of the filtrate obtained in Assay Weigh accurately about 0.5 g of Azathioprine, pre-
(2) add 1 mL of dilute hydrochloric acid and water to make viously dried, add 80 mL of N, N-dimethylformamide, and
50 mL, and perform the test using this solution as the test so- warm to dissolve. After cooling, titrate <2.50> with 0.1
lution. Prepare the control solution with 0.40 mL of 0.005 mol/L tetramethylammonium hydroxide VS until the color
mol/L sulfuric acid VS (not more than 0.038z). of the solution changes from yellow through yellow-green to
(4) Heavy metals <1.07>—Proceed with 2.0 g of blue-green (indicator: 1 mL of thymol blue-dimethylfor-
Azathioprine according to Method 2, and perform the test. mamide TS). Perform a blank determination, and make any
Prepare the control solution with 2.0 mL of Standard Lead necessary correction.
Each mL of 0.1 mol/L tetramethylammonium hydroxide VS
＝ 27.73 mg of C9H7O7O2S
of Azathioprine, according to Method 3, and perform the
test (not more than 2 ppm). Containers and storage Containers—Well-closed contain-
(6) Related substances—Dissolve 10 mg of Azathioprine ers.
in 80 mL of the mobile phase by warming, cool, add the Storage—Light-resistant.
mobile phase to make 100 mL, and use this solution as the
water to make exactly 100 mL, and use this solution as the Azathioprine Tablets
standard solution. Perform the test with exactly 20 mL each
of the sample solution and standard solution as directed アザチオプリン錠
lowing conditions. Determine each peak area by the auto-
Azathioprine Tablets contain not less than 95.0z
matic integration method: the total area of the peaks other
than that of azathioprine from the sample solution is not
azathioprine (C9H7N7O2S: 277.26).
larger than 1/2 times the peak area of azathioprine from the
standard solution. Method of preparation Prepare as directed under Tablets,
Operating conditions— with Azathioprine.
Identification (1) Weigh a quantity of powdered
length: 296 nm).
Azathioprine Tablets, equivalent to 0.01 g of Azathioprine.
Add 50 mL of water, shake well while warming, and filter.
Proceed with 5 mL of the filtrate as directed in the Identifi-
cation (1) under Azathioprine.
(2) Proceed with 1 mL of the filtrate obtained in (1) as
409 C.
directed in the Identification (2) under Azathioprine.
Mobile phase: Adjust to pH 2.5 of a solution of 0.05
(3) Determine the absorption spectrum of the sample so-
mol/L potassium dihydrogenphosphate TS (1 in 2) with
lution in the Assay as directed under Ultraviolet-visible Spec-
diluted phosphoric acid (3 in 2000). To 800 mL of this solu-
trophotometry <2.24>: it exhibits a maximum between 278
tion add 200 mL of methanol.
nm and 282 nm.
Flow rate: Adjust so that the retention time of
(4) Weigh a quantity of powdered Azathioprine Tablets,
azathioprine is about 8 minutes.
equivalent to 0.1 g of Azathioprine to the labeled amount.
JP XVII Official Monographs / Azelastine Hydrochloride 461
Add 10 mL of a solution of ammonia solution (28) in metha- of the subsequent filtrate, add 0.1 mol/L hydrochloric acid
nol (1 in 10), shake well, filter, and use the filtrate as the TS to make exactly 100 mL, and use this solution as the sam-
sample solution. Separately, dissolve 0.1 g of Azathioprine ple solution. Separately, weigh accurately about 0.1 g of
RS in 10 mL of a solution of ammonia solution (28) in meth- Azathioprine RS, previously dried at 1059C for 5 hours, dis-
anol (1 in 10), and use this solution as the standard solution. solve in 20 mL of dimethylsulfoxide for ultraviolet-visible
Perform the test with these solutions as directed under Thin- spectrophotometry, and add 0.1 mol/L hydrochloric acid TS
layer Chromatography <2.03>. Spot 5 mL each of the sample to make exactly 500 mL. Measure exactly 3 mL of this solu-
solution and standard solution on a plate of silica gel with tion, add 0.1 mol/L hydrochloric acid TS to make exactly
fluorescent indicator for thin-layer chromatography. De- 100 mL, and use this solution as the standard solution. De-
velop the plate with a mixture of chloroform, a solution of termine the absorbances, AT and AS, of the sample solution
ammonia solution (28) in methanol (1 in 10), n-butyl for- and standard solution at 280 nm as directed under Ultravio-
mate and 1,2-dichloroethane (15:10:5:2) to a distance of let-visible Spectrophotometry <2.24>.
Amount (mg) of azathioprine (C9H7N7O2S)
let light (main wavelength: 254 nm): the spots from the sam-
＝ M S × AT / AS
ple solution and the standard solution show the same R f
value. MS: Amount (mg) of Azathioprine RS taken
Uniformity of dosage units <6.02> Perform the Mass varia- Containers and storage Containers—Tight containers.
tion test, or the Content uniformity test according to the fol- Storage—Light-resistant.
To 1 tablet of Azathioprine Tablets add 1 mL of dimethyl-
sulfoxide for ultraviolet-visible spectrophotometry per 5 mg Azelastine Hydrochloride
of azathioprine (C9H7N7O2S), shake well, add 0.1 mol/L hy-
drochloric acid TS to make exactly V mL so that each mL アゼラスチン塩酸塩
contains about 0.2 mg of azathioprine (C9H7N7O2S), and
filter. Discard the first 20 mL of the filtrate, pipet 3 mL of
the subsequent filtrate, add 0.1 mol/L hydrochloric acid TS
to make exactly 100 mL, and use this solution as the sample
Amount (mg) of azathioprine (C9H7N7O2S)
＝ MS × AT/AS × V/500
MS: Amount (mg) of Azathioprine RS taken
Dissolution <6.10> When the test is performed at 50 revolu- C22H24ClN3O.HCl: 418.36
tions per minute according to the Paddle method, using 900 4-[(4-Chlorophenyl)methyl]-2-[(4RS )-
mL of water as the dissolution medium, the dissolution rate (1-methylazepan-4-yl)]phthalazin-1(2H )-one
in 45 minutes of Azathioprine Tablets is not less than 80z. monohydrochloride
Start the test with 1 tablet of Azathioprine Tablets, with- [79307-93-0]
minute after starting the test, and filter through a membrane Azelastine Hydrochloride, when dried, contains
filter with a pore size not exceeding 0.8 mm. Discard the first not less than 99.0z and not more than 101.0z of
10 mL of the filtrate, pipet V mL of the subsequent filtrate, azelastine hydrochloride (C22H24ClN3O.HCl).
add water to make exactly V? mL so that each mL contains
Description Azelastine Hydrochloride occurs as a white
about 11 mg of azathioprine (C9H7N7O2S), and use this solu-
crystalline powder.
tion as the sample solution. Separately, weigh accurately
It is freely soluble in formic acid, and slightly soluble in
about 10 mg of Azathioprine RS, previously dried at 1059 C
water and in ethanol (99.5).
for 5 hours, and dissolve in water to make exactly 100 mL.
A solution of Azelastine Hydrochloride (1 in 200) shows
mine the absorbances, AT and AS, at 280 nm of the sample
solution and standard solution as directed under Ultraviolet- Identification (1) Determine the absorption spectrum of a
visible Spectrophotometry <2.24>. solution of Azelastine Hydrochloride (3 in 100,000) as di-
rected under Ultraviolet-visible Spectrophotometry <2.24>,
of azathioprine (C9H7N7O2S)
＝ MS × AT/AS × V?/V × 1/C × 108
same wavelengths.
MS: Amount (mg) of Azathioprine RS taken (2) Determine the infrared absorption spectrum of
C: Labeled amount (mg) of azathioprine (C9H7N7O2S) in 1 Azelastine Hydrochloride as directed in the potassium chlo-
tablet ride disk method under Infrared Spectrophotometry <2.25>:
same wave numbers.
Azathioprine Tablets. Weigh accurately a portion of the
(3) To 10 mL of a saturated solution of Azelastine
powder, equivalent to about 0.1 g of azathioprine
Hydrochloride add 1 mL of dilute nitric acid, and filter to
(C9H7N7O2S), add 20 mL of dimethylsulfoxide for ultravio-
separate formed crystals: the filtrate responds to the Qualita-
let-visible spectrophotometry, shake well, add 0.1 mol/L
tive Tests <1.09> (2) for chloride.
hydrochloric acid TS to make exactly 500 mL, and filter.
Discard the first 20 mL of the filtrate, measure exactly 3 mL Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
462 Azelastine Hydrochloride Granules / Official Monographs JP XVII
Azelastine Hydrochloride according to Method 2, and per-
form the test. Prepare the control solution with 2.0 mL of Azelastine Hydrochloride Granules
(2) Arsenic <1.11>—Prepare the test solution with 1.0 g アゼラスチン塩酸塩顆粒
of Azelastine Hydrochloride according to Method 3, and
Azelastine Hydrochloride Granules contain not
(3) Related substances—Dissolve 50 mg of Azelastine
the labeled amount of azelastine hydrochloride
(C22H24ClN3O.HCl: 418.36).
use this solution as the standard solution. Perform the test Method of preparation Prepare as directed under Gran-
with exactly 20 mL each of the sample solution and standard ules, with Azelastine Hydrochloride.
Identification To a quantity of Azelastine Hydrochloride
Granules, equivalent to 2 mg of Azelastine Hydrochloride,
area of these solutions by the automatic integration method:
add 30 mL of 0.1 mol/L hydrochloric acid TS, and treat
each peak area other than azelastine obtained from the sam-
with ultrasonic waves for 30 minutes. After cooling, add 0.1
ple solution is not larger than 1/10 times the peak area of
mol/L hydrochloric acid TS to make 50 mL, and centrifuge.
azelastine obtained from the standard solution, and the total
Determine the absorption spectrum of the supernatant liquid
area of the peaks other than the peak of azelastine from the
as directed under Ultraviolet-visible Spectrophotometry
sample solution is not larger than 1/2 times the peak area of
<2.24>: it exhibits a maximum between 283 nm and 287 nm.
azelastine from the standard solution.
Operating conditions— Dissolution <6.10> When the test is performed at 50 revolu-
Detector: An ultraviolet absorption photometer (wave- tions per minute according to the Paddle method, using 900
length: 240 nm). mL of 0.05 mol/L acetic acid-sodium acetate buffer solution
Column: A stainless steel column 4.6 mm in inside diame- (pH 4.0) as the dissolution medium, the dissolution rate in
ter and 15 cm in length, packed with octadecylsilanized silica 45 minutes of Azelastine Hydrochloride Granules is not less
gel for liquid chromatography (5 mm in particle diameter). than 80z.
Column temperature: A constant temperature of about Start the test with accurately weighed amount of
359 C. Azelastine Hydrochloride Granules, equivalent to about
Mobile phase: A mixture of water, acetonitrile and per- 1 mg of azelastine hydrochloride (C22H24ClN3O.HCl), with-
chloric acid (660:340:1). draw not less than 20 mL of the medium at the specified
Flow rate: Adjust so that the retention time of azelastine is minute after starting the test, and filter through a membrane
about 10 minutes. filter with a pore size not exceeding 0.5 mm. Discard the first
Time span of measurement: About 2 times as long as the 10 mL of the filtrate, and use the subsequent filtrate as the
retention time of azelastine, beginning after the solvent sample solution. Separately, weigh accurately about 50 mg
peak. of azelastine hydrochloride for assay, previously dried at
System suitability— 1059C for 2 hours, and dissolve in the dissolution medium to
Test for required detectability: Pipet 5 mL of the standard make exactly 250 mL. Pipet 1 mL of this solution, add the
solution, and add the mobile phase to make exactly 50 mL. dissolution medium to make exactly 200 mL, and use this so-
Confirm that the peak area of azelastine obtained from 20 lution as the standard solution. Perform the test with exactly
mL of this solution is equivalent to 7 to 13z of that obtained 50 mL each of the sample solution and standard solution as
from 20 mL of the standard solution. directed under Liquid Chromatography <2.01> according to
System performance: When the procedure is run with 20 the following conditions, and determine the peak areas, AT
mL of the standard solution under the above operating con- and AS, of azelastine in each solution.
factor of the peak of azelastine is not less than 5000 and not
of azelastine hydrochloride (C22H24ClN3O.HCl)
＝ MS/MT × AT/AS × 1/C × 9/5
with 20 mL of the standard solution under the above operat- MS: Amount (mg) of azelastine hydrochloride for assay
ing conditions, the relative standard deviation of the peak taken
area of azelastine is not more than 1.0z. MT: Amount (g) of Azelastine Hydrochloride Granules
taken
C: Labeled amount (mg) of azelastine hydrochloride
2 hours).
(C22H24ClN3O.HCl) in 1 g
Assay Weigh accurately about 0.6 g of previously dried Proceed as directed in the operating conditions in the
Azelastine Hydrochloride, dissolve in 5 mL of formic acid, Assay.
add 70 mL of acetic anhydride, and titrate <2.50> with 0.1 System suitability—
mol/L perchloric acid VS (potentiometric titration). Per- System performance: When the procedure is run with 50
form a blank determination in the same manner, and make mL of the standard solution under the above operating con-
any necessary correction. ditions, the number of theoretical plates and the symmetry
factor of the peak of azelastine are not less than 2000 and
＝ 41.84 mg of C22H24ClN3O.HCl
Containers and storage Containers—Tight containers. with 50 mL of the standard solution under the above operat-
Storage—Light-resistant. ing conditions, the relative standard deviation of the peak
JP XVII Official Monographs / Azelnidipine 463
area of azelastine is not more than 2.0z.

Assay Weigh accurately an amount of Azelastine Hydro- Azelnidipine
chloride Granules, equivalent to about 2 mg of azelastine hy-
アゼルニジピン
drochloride (C22H24ClN3O.HCl), add 50 mL of 0.1 mol/L
hydrochloric acid TS, treat with ultrasonic waves for 20
minutes, add 40 mL of ethanol (99.5), add exactly 5 mL of
the internal standard solution, and add ethanol (99.5) to
make 100 mL. Centrifuge this solution, and use the superna-
tant liquid as the sample solution. Separately, weigh accu-
rately about 50 mg of azelastine hydrochloride for assay,
previously dried at 1059C for 2 hours, and dissolve in water
to make exactly 50 mL. Pipet 10 mL of this solution, and
C33H34N4O6: 582.65
add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL.
3-[1-(Diphenylmethyl)azetidin-3-yl] 5-(1-methylethyl)
Pipet 10 mL of this solution, add 40 mL of 0.1 mol/L hydro-
(4RS)-2-amino-6-methyl-4-(3-nitrophenyl)-1,4-dihydropyridine-
chloric acid TS and 40 mL of ethanol (99.5), add exactly 5
3,5-dicarboxylate
mL of the internal standard solution, add ethanol (99.5) to
[123524-52-7]
make 100 mL, and use this solution as the standard solution.
Perform the test with 20 mL each of the sample solution and
Azelnidipine contains not less than 99.0z and not
standard solution as directed under Liquid Chromatography
<2.01> according to the following conditions, and calculate
more than 101.0z of azelnidipine (C33H34N4O6), cal-
culated on the dried basis.
the ratios, QT and QS, of the peak area of azelastine to that
of the internal standard. Description Azelnidipine occurs as a light yellow to yellow,
crystalline powder or powder containing masses.
Amount (mg) of azelastine hydrochloride
It is freely soluble in ethanol (99.5) and in acetic acid
(C22H24ClN3O.HCl)
(100), and practically insoluble in water.
＝ MS × QT/QS × 1/25
A solution of Azelnidipine in ethanol (99.5) (1 in 100)
MS: Amount (mg) of azelastine hydrochloride for assay shows no optical rotation.
taken Azelnidipine shows crystal polymorphism.
Internal standard solution—Dissolve 0.2 g of 2-ethylhexyl Identification (1) Determine the absorption spectrum of a
parahydroxybenzoate in ethanol (99.5) to make 100 mL. solution of Azelnidipine in ethanol (99.5) (1 in 50,000) as
Operating conditions— directed under Ultraviolet-visible Spectrophotometry <2.24>,
Detector: An ultraviolet absorption photometer (wave- and compare the spectrum with the Reference Spectrum:
length: 285 nm). both spectra exhibit similar intensities of absorption at the
Column: A stainless steel column 4.6 mm in inside diame- same wavelengths.
ter and 15 cm in length, packed with octadecylsilanized silica (2) Determine the infrared absorption spectrum of Azel-
gel for liquid chromatography (5 mm in particle diameter). nidipine as directed in the paste method under Infrared Spec-
Column temperature: A constant temperature of about trophotometry <2.25>, and compare the spectrum with the
409 C. Reference Spectrum: both spectra exhibit similar intensities
Mobile phase: A mixture of acetonitrile and a solution of of absorption at the same wave numbers.
sodium lauryl sulfate in diluted acetic acid (100) (1 in 250)
(1 in 500) (11:9).
Azelnidipine according to Method 2, and perform the test.
Flow rate: Adjust so that the retention time of azelastine is
about 6 minutes.
(2) Related substances—Dissolve 0.10 g of Azelnidipine
in a mixture of acetonitrile and water (4:1) to make 100 mL,
and use this solution as the sample solution. Pipet 2 mL of
ditions, azelastine and the internal standard are eluted in this
the sample solution, add a mixture of acetonitrile and water
(4:1) to make exactly 200 mL, and use this solution as the
than 2.0.
the peak area of azelastine to that of the internal standard is
matic integration method: the areas of the peak, having the
not more than 1.0z.
relative retention time of about 0.50 and about 1.42 to azel-
Containers and storage Containers—Tight containers. nidipine, obtained from the sample solution are not larger
than 1/5 times and 3/10 times the peak area of azelnidipine
obtained from the standard solution, respectively, the area
of the peak other than azelnidipine and the peaks mentioned
above from the sample solution is not larger than 1/10 times
the peak area of azelnidipine from the standard solution,
and the total area of the peaks other than azelnidipine from
the sample solution is not larger than 7/10 times the peak
area of azelnidipine from the standard solution.
464 Azelnidipine Tablets / Official Monographs JP XVII
length: 220 nm). tween 253 nm and 257 nm and between 339 nm and 346 nm.
Purity Related substances—Conduct this procedure using
light-resistant vessels. Powder Azelnidipine Tablets. Weigh a
portion of the powder, equivalent to 10 mg of Azelnidipine,
add 10 mL of a mixture of acetonitrile and water (4:1),
409 C.
agitate gently, then disperse to fine particles with the aid of
ultrasonic waves for 15 minutes. Centrifuge this solution,
phosphate in 350 mL of water, add 650 mL of a mixture of
and use the supernatant liquid as the sample solution. Pipet
acetonitrile and methanol (7:3), and adjust to pH 5.5 with
2 mL of the sample solution, add a mixture of acetonitrile
diluted phosphoric acid (1 in 10).
and water (4:1) to make exactly 100 mL, and use this solu-
Flow rate: Adjust so that the retention time of azelnidi-
tion as the standard solution. Perform the test with exactly
pine is about 36 minutes.
retention time of azelnidipine, beginning after the solvent
the following conditions, and determine each peak area by
peak.
the automatic integration method: the areas of the peaks,
having the relative retention times of about 0.10, about 0.13,
about 0.50, and about 1.42 to azelnidipine, obtained from
standard solution add a mixture of acetonitrile and water
the sample solution, are not larger than 9/20 times, 1/5
(4:1) to make exactly 20 mL. Confirm that the peak area of
times, 2/5 times, and 2/5 times the peak area of azelnidipine
azelnidipine obtained with 10 mL of this solution is equiva-
obtained from the standard solution, respectively, the area
lent to 3.5 to 6.5z of that with 10 mL of the standard solu-
of the peak, other than azelnidipine and the peaks men-
tion.
tioned above, is not larger than 1/10 times the peak area of
azelnidipine from the standard solution. Furthermore, the
total area of these peaks other than azelnidipine is not larger
than 1.75 times the peak area of azelnidipine from the stand-
factor of the peak of azelnidipine are not less than 15,000
ard solution.
and 0.8 to 1.5, respectively.
the Purity (2) under Azelnidipine.
area of azelnidipine is not more than 1.0z.
Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- retention time of azelnidipine.
um, 709C, 5 hours). System suitability—
Assay Weigh accurately about 0.4 g of Azelnidipine, dis- (4:1) to make exactly 20 mL. Confirm that the peak area of
solve in 50 mL of acetic acid (100), and titrate <2.50> with azelnidipine obtained with 10 mL of this solution is equiva-
0.1 mol/L perchloric acid VS (potentiometric titration). Per- lent to 3.5 to 6.5z of that obtained with 10 mL of the stand-
form a blank determination in the same manner, and make ard solution.
any necessary correction. System performance: When the procedure is run with 10
＝ 29.13 mg of C33H34N4O6
factor of the peak of azelnidipine are not less than 15,000
Containers and storage Containers—Tight containers. and not more than 1.5, respectively.
Azelnidipine Tablets ing conditions, the relative standard deviation of the peak
area of azelnidipine is not more than 1.0z.
アゼルニジピン錠
Azelnidipine Tablets contain not less than 95.0z Content uniformity test.
and not more than 105.0z of the labeled amount of To 1 tablet of Azelnidipine Tablets add exactly 1 mL of
azelnidipine (C33H34N4O6: 582.65). the internal standard solution per 2 mg of azelnidipine
(C33H34N4O6), and add a mixture of acetonitrile and water
(4:1) to make 32 mL. Disintegrate the tablet with occasional
with Azelnidipine.
shaking, and treat with ultrasonic waves for 10 minutes.
Identification Powder Azelnidipine Tablets. Weigh a por- Centrifuge this solution, pipet V mL of the supernatant liq-
tion of the powder, equivalent to 4 mg of Azelnidipine, add uid, equivalent to 2.5 mg of azelnidipine (C33H34N4O6), add
150 mL of ethanol (99.5), treat with ultrasonic waves for a mixture of acetonitrile and water (4:1) to make 50 mL, and
15 minutes, then add ethanol (99.5) to make 200 mL. Centri- use this solution as the sample solution. Then, proceed as di-
fuge this solution, filter the supernatant liquid through a rected in the Assay.
glass wool filter with a pore size not exceeding 0.7 mm. Dis-
Amount (mg) of azelnidipine (C33H34N4O6)
card the first 10 mL of the filtrate, and use the subsequent
＝ MS × QT/QS × 8/5V
filtrate as the sample solution. Determine the absorption
spectrum of the sample solution as directed under Ultravio- MS: Amount (mg) of azelnidipine for assay taken
let-visible Spectrophotometry <2.24>: it exhibits maxima be-
JP XVII Official Monographs / Azithromycin Hydrate 465
Internal standard solution—A solution of 2,2?-dinaphthyl- Mobile phase: Dissolve 0.9 g of potassium dihydrogen
ether in a mixture of acetonitrile and water (4:1) (1 in 1000). phosphate in 300 mL of water, add 700 mL of acetonitrile,
then adjust to pH 6.0 with dilute sodium hydroxide TS.
Flow rate: Adjust so that the retention time of azelnidi-
pine is about 13 minutes.
900 mL of 1st fluid for dissolution test as the dissolution
medium, the dissolution rate in 45 minutes of Azelnidipine
Start the test with 1 tablet of Azelnidipine Tablets, with-
ditions, azelnidipine and the internal standard are eluted in
less than 12.
first 10 mL of the filtrate, pipet V mL of the subsequent fil-
trate, add the dissolution medium to make exactly V? mL
so that each mL contains about 8.9 mg of azelnidipine
the peak area of azelnidipine to that of the internal standard
(C33H34N4O6), and use this solution as the sample solution.
Separately, weigh accurately about 45 mg of azelnidipine for
assay, previously dried in vacuum at 709C for 5 hours, dis- Containers and storage Containers—Tight containers.
solve in ethanol (99.5) to make exactly 25 mL. Pipet 1 mL of Storage—Light-resistant.
this solution, add the dissolution medium to make exactly
200 mL, and use this solution as the standard solution. De-
termine the absorbances, AT and AS, at 270 nm of the sam- Azithromycin Hydrate
ple solution and standard solution as directed under Ultravi-
olet-visible Spectrophotometry <2.24>, using the dissolution アジスロマイシン水和物
medium as the blank.
of azelnidipine (C33H34N4O6)
＝ MS × AT/AS × V?/V × 1/C × 18
MS: Amount (mg) of azelnidipine for assay taken
C: Labeled amount (mg) of azelnidipine (C33H34N4O6) in 1
tablet
Assay Weigh accurately the mass of not less than 20 Azel-
nidipine Tablets, and powder. Weigh accurately a portion of
the powder, equivalent to about 50 mg of azelnidipine
(C33H34N4O6), add exactly 25 mL of the internal standard so-
lution, add 50 mL of a mixture of acetonitrile and water
C38H72N2O12.2H2O: 785.02
(4:1). After treating with ultrasonic waves for 10 minutes,
(2R,3S,4S,5R,6R,8R,11R,12R,13S,14R)-5-
add a mixture of acetonitrile and water (4:1) to make 100
(3,4,6-Trideoxy-3-dimethylamino-b-D-xylo-
mL. Centrifuge this solution, to 5 mL of the supernatant liq-
hexopyranosyloxy)-3-(2,6-dideoxy-3-
uid add a mixture of acetonitrile and water (4:1) to make 50
C-methyl-3-O-methyl-a-L-ribo-hexopyranosyloxy)-
10-aza-6,12,13-trihydroxy-2,4,6,8,10,11,13-
weigh accurately about 50 mg of azelnidipine for assay, pre-
heptamethylhexadecan-14-olide dihydrate
viously dried in vacuum at 709C for 5 hours, dissolve in ex-
[117772-70-0]
actly 25 mL of the internal standard solution, and add a mix-
ture of acetonitrile and water (4:1) to make 100 mL. To 5
Azithromycin Hydrate is the derivative of
mL of this solution add a mixture of acetonitrile and water
erythromycin.
(4:1) to make 50 mL, and use this solution as the standard
solution. Perform the test with 10 mL each of the sample so-
lution and standard solution as directed under Liquid Chro-
anhydrous basis. The potency of Azithromycin Hy-
drate is expressed as mass (potency) of azithromycin
and calculate the ratios, QT and QS, of the peak area of azel-
(C38H72N2O12: 748.98).
nidipine to that of the internal standard.
Description Azithromycin Hydrate occurs as a white crys-
Amount (mg) of azelnidipine (C33H34N4O6) ＝ MS × QT/QS
talline powder.
MS: Amount (mg) of azelnidipine for assay taken It is freely soluble in methanol and in ethanol (99.5), and
Internal standard solution—2,2?-dinaphthylether in a mix-
ture of acetonitrile and water (4:1) (1 in 1000). Identification Determine the infrared absorption spectrum
Operating conditions— of Azithromycin Hydrate as directed in the potassium bro-
Detector: An ultraviolet absorption photometer (wave- mide disk method under the Infrared Spectrophotometry
length: 254 nm). <2.25>, and compare the spectrum with the Reference Spec-
Column: A stainless steel column 4.6 mm in inside diame- trum or the spectrum of Azithromycin RS: both spectra
ter and 25 cm in length, packed with octadecylsilanized silica exhibit similar intensities of absorption at the same wave
gel for liquid chromatography (5 mm in particle diameter). numbers.
D : －45 – －499(0.4 g calculated
409 C.
on the anhydrous basis, ethanol (99.5), 20 mL, 100 mm).
466 Aztreonam / Official Monographs JP XVII
Azithromycin Hydrate according to Method 2, and perform Aztreonam
ard Lead Solution (not more than 10 ppm). アズトレオナム
(2) Related substances—Being specified separately when
the drug is granted approval based on the Law.
(0.4 g, volumetric titration, direct titration).
Assay Weigh accurately an amount of Azithromycin Hy-
drate and Azithromycin RS, equivalent to about 50 mg (po- C13H17N5O8S2: 435.43
tency), dissolve each in an adequate amount of a mixture of 2-{(Z )-(2-Aminothiazol-4-yl)-[(2S,3S )-2-methyl-
acetonitrile and water (3:2), add exactly 2 mL of the internal 4-oxo-1-sulfoazetidin-3-ylcarbamoyl]methyleneaminooxy}-
standard solution and the mixture of acetonitrile and water 2-methyl-1-propanoic acid
(3:2) to make 50 mL, and use these solutions as the sample [78110-38-0]
solution and standard solution. Perform the test with 5 mL
each of the sample solution and standard solution as directed Aztreonam contains not less than 920 mg (potency)
under Liquid Chromatography <2.01> according to the fol- and not more than 1030 mg (potency) per mg, calcu-
lowing conditions, and calculate the ratios, QT and QS, of lated on the anhydrous basis. The potency of Aztreo-
the peak area of azithromycin to that of the internal stand- nam is expressed as mass (potency) of aztreonam
ard. (C13H17N5O8S2).
Amount [ mg (potency)] of azithromycin (C38H72N2O12) Description Aztreonam occurs as a white to yellowish
＝ MS × QT/QS × 1000 white crystalline powder.
It is freely soluble in dimethylsulfoxide, slightly soluble in
MS: Amount [mg (potency)] of Azithromycin RS taken
water and in methanol, and very slightly soluble in ethanol
Internal standard solution—A solution of 4,4?- (95).
bis(diethylamino)benzophenone in acetonitrile (3 in 4000).
solution of Aztreonam (3 in 100,000) as directed under Ul-
length: 215 nm).
spectrum with the Reference Spectrum or the spectrum of a
solution of Aztreonam RS prepared in the same manner as
ter and 25 cm in length, packed with octadecylsilanized poly-
the sample solution: both spectra exhibit similar intensities
vinyl alcohol gel polymer for liquid chromatography (5 mm
of absorption at the same wavelengths.
in particle diameter).
(2) Determine the spectrum of a solution of Aztreonam
in deuterated dimethylsulfoxide for nuclear magnetic reso-
409 C.
nance spectroscopy (1 in 10), using a light hydrogen sub-
Mobile phase: Dissolve 6.97 g of dipotassium hydrogen
stance existing in deuterated dimethylsulfoxide for nuclear
phosphate in about 750 mL of water, adjust the pH to 11.0
magnetic resonance spectroscopy as an internal reference
with potassium hydroxide TS, and add water to make 1000
compound and 2.50 ppm for its chemical shift, as directed
mL. To 400 mL of this solution add 600 mL of acetonitrile
under Nuclear Magnetic Resonance Spectroscopy <2.21>
for liquid chromatography.
(1H): it exhibits a multiple signal at around d 1.5 ppm, and a
Flow rate: Adjust so that the retention time of azithromy-
single signal at around d 7.0 ppm. The ratio of integrated in-
cin is about 10 minutes.
tensity of each signal is 9:1.
System performance: When the procedure is run with 5 mL Optical rotation <2.49> [a]20
D : －26 – －329 (0.25 g calcu-
of the standard solution under the above operating condi- lated on the anhydrous bases, water, 50 mL, 100 mm).
tions, azithromycin and the internal standard are eluted in
pH <2.54> Dissolve 0.05 g of Aztreonam in 10 mL of
water: the pH of this solution is between 2.2 and 2.8.
less than 2.0.
System repeatability: When the test is repeated 6 times Purity (1) Clarity and color of solution—Dissolve 1.0 g
with 5 mL of the standard solution under the above operating of Aztreonam in 20 mL of dimethylsulfoxide: the solution is
conditions, the relative standard deviation of the ratios of clear, and its absorbance at 420 nm, determined as directed
the peak area of azithromycin to that of the internal stand- under Ultraviolet-visible Spectrophotometry <2.24>, is not
ard is not more than 1.0z. more than 0.06.
(2) Heavy metals <1.07>—Proceed with 2.0 g of Aztreo-
nam according to Method 2, and perform the test. Prepare
the control solution with 2.0 mL of Standard Lead Solution
(3) Related substances—Dissolve 40 mg of Aztreonam in
100 mL of water, and use this solution as the sample solu-
tion. Pipet 2 mL of the sample solution, add water to make
exactly 100 mL, and use this solution as the standard solu-
tion. Perform the test with exactly 25 mL each of the sample
JP XVII Official Monographs / Aztreonam for Injection 467
tions, and determine each peak area of both solutions by the methanol.
automatic integration method: the area of the peak other Flow rate: Adjust so that the retention time of aztreonam
than aztreonam obtained from the sample solution is not is about 8 minutes.
larger than the peak area of aztreonam from the standard System suitability—
solution, and the total area of peaks other than aztreonam System performance: When the procedure is run with 25
from the sample solution is not larger than 2.5 times the mL of the standard solution under the above operating con-
peak area of aztreonam from the standard solution. ditions, the internal standard and aztreonam are eluted in
Operating conditions— this order with the resolution between these peaks being not
Column, column temperature, mobile phase, and flow less than 4.
rate: Proceed as directed in the operating conditions in the System repeatability: When the test is repeated 6 times
Assay. with 25 mL of the standard solution under the above operat-
Detector: An ultraviolet absorption photometer (wave- ing conditions, the relative standard deviation of the ratios
length: 254 nm). of the peak area of aztreonam to that of the internal stand-
Time span of measurement: About 4 times as long as the ard is not more than 1.5z.
retention time of aztreonam, beginning after the solvent
peak.
Test for required detectability: To 5 mL of the standard
solution add water to make 10 mL, and use this solution as
the solution for system suitability test. Pipet 1 mL of the so- Aztreonam for Injection
lution for system suitability test, and add water to make ex-
注射用アズトレオナム
actly 10 mL. Confirm that the peak area of aztreonam ob-
tained from 25 mL of this solution is equivalent to 7 to 13z
of that obtained from 25 mL of the solution for system suita- Aztreonam for Injection is a preparation for injec-
bility test. tion which is dissolved before use.
System performance: When the procedure is run under the It contains not less than 93.0z and not more
above operating conditions with 25 mL of the standard solu- than 107.0z of the labeled potency of aztreonam
tion obtained in the Assay, the internal standard and aztreo- (C13H17N5O8S2: 435.43).
nam are eluted in this order with the resolution between
tions, with Aztreonam.
with 25 mL of the standard solution under the above operat- Description Aztreonam for Injection is white to yellowish
ing conditions, the relative standard deviation of the peak white masses or powder.
areas of aztreonam is not more than 2.0z.
Identification (1) Dissolve an amount of Aztreonam for
Water <2.48> Not more than 2.0z (0.5 g, volumetric titra- Injection, equivalent to 6 mg (potency) of Aztreonam, in
tion, direct titration). 1 mL of hydroxylammonium chloride-ethanol TS, allow to
stand for 3 minutes, add 1 mL of acidic ammonium iron
(III) sulfate TS, and mix: a red-brown color develops.
Assay Weigh accurately an amount of Aztreonam and Az- (2) Dissolve an amount of Aztreonam for Injection,
treonam RS, equivalent to about 20 mg (potency), dissolve equivalent to 3 mg (potency) of Aztreonam, in 100 mL of
each in 70 mL of water, add exactly 10 mL of the internal water, and determine the absorption spectrum of the solu-
standard solution and water to make 100 mL, and use these tion as directed under Ultraviolet-visible Spectrophotometry
solutions as the sample solution and standard solution, re- <2.24>: it exhibits a maximum between 289 nm and 293 nm.
spectively. Perform the test with 25 mL each of these solu-
tions as directed under Liquid Chromatography <2.01> ac-
amount of Aztreonam for Injection, equivalent to 1.0 g
(potency) of Aztreonam, in 10 mL of water is 4.5 to 7.0.
QT and QS, of the peak area of aztreonam to that of the in-
ternal standard. Purity Clarity and color of solution—Dissolve an amount
of Aztreonam for Injection, equivalent to 1.0 g (potency) of
Amount [ mg (potency)] of aztreonam (C13H17N5O8S2)
Aztreonam, in 10 mL of water: the solution is clear, and its
＝ MS × QT/QS × 1000
absorbance <2.24> at 450 nm is not more than 0.06.
MS: Amount [mg (potency)] of Aztreonam RS taken
Internal standard solution—A solution of 4-aminobenzoic tion, direct titration).
acid (1 in 6250).
tency).
length: 280 nm). Uniformity of dosage units <6.02> It meets the requirement
Column: A stainless steel column 4.6 mm in inside diame- of the Mass variation test.
409 C. Insoluble particulate matter <6.07> It meets the require-
Mobile phase: Dissolve 1.7 g of tetrabutylammonium ment.
hydrogensulfate in 300 mL of water, adjust to pH 3.0 with
0.5 mol/L disodium hydrogenphosphate TS, and add water
to make 1000 mL. To 650 mL of this solution add 350 mL of
468 Bacampicillin Hydrochloride / Official Monographs JP XVII
Assay Take an amount of Aztreonam for Injection, Bacampicillin Hydrochloride as directed in the potassium
equivalent to about 5 g (potency) of Aztreonam, dissolve the chloride disk method under Infrared Spectrophotometry
contents with a suitable amount of water, and transfer to a <2.25>, and compare the spectrum with the Reference Spec-
100-mL volumetric flask. Wash each container with water, trum or the spectrum of Bacampicillin Hydrochloride RS:
combine the washings and the solution, and add water to both spectra exhibit similar intensities of absorption at the
make exactly 100 mL. Pipet 10 mL of this solution, and add same wave numbers.
water to make exactly 50 mL. Pipet 2 mL of this solution, (3) A solution of Bacampicillin Hydrochloride (1 in 50)
add exactly 10 mL of the internal standard solution and responds to the Qualitative Tests <1.09> for chloride.
water to make 100 mL, and use this solution as the sample
D : ＋140 – ＋1709(0.1 g calcu-
solution. Separately, weigh accurately an amount of Aztreo-
lated on the anhydrous basis, ethanol (95), 25 mL, 100 mm).
nam RS, equivalent to about 20 mg (potency), dissolve in a
suitable amount of water, add exactly 10 mL of the internal Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
standard solution and water to make 100 mL, and use this Bacampicillin Hydrochloride according to Method 2, and
solution as the standard solution. Then, proceed as directed perform the test. Prepare the control solution with 2.0 mL of
in the Assay under Aztreonam. Standard Lead Solution (not more than 20 ppm).
Amount [mg (potency)] of aztreonam (C13H17N5O8S2)
of Bacampicillin Hydrochloride according to Method 3, and
＝ MS × QT/QS × 250
MS: Amount [mg (potency)] of Aztreonam RS taken (3) Free ampicillin—Carry out the determination imme-
diately after preparing the sample solution. Weigh accurately
Internal standard solution—A solution of 4-aminobenzoic
about 0.1 g of Bacampicillin Hydrochloride, dissolve in ex-
acid (1 in 6250).
actly 10 mL of the internal standard solution, add the mobile
Containers and storage Containers—Hermetic containers. phase to make 20 mL, and use this solution as the sample so-
Storage—Light-resistant. lution. Separately, weigh accurately an amount of Ampicil-
lin RS, equivalent to about 25 mg (potency), and dissolve in
water to make exactly 100 mL. Pipet 4 mL of this solution,
Bacampicillin Hydrochloride add exactly 10 mL of the internal standard solution, add the
mobile phase to make 20 mL, and use this solution as the
Ampicillin Ethoxycarbonyloxyethyl standard solution. Perform the test with 10 mL each of the
Hydrochloride
バカンピシリン塩酸塩 ditions, and calculate the ratios, QT and QS, of the peak area
of ampicillin to that of the internal standard in each solu-
tion. The amount of ampicillin, calculated by the following
equation, is not more than 1.0z.
Amount (z) of ampicillin (C16H19N3O4S)
＝ M S / M T × QT / QS × 4
MS: Amount [mg (potency)] of Ampicillin RS taken
MT: Amount (mg) of Bacampicillin Hydrochloride taken
C21H27N3O7S.HCl: 501.98
1-Ethoxycarbonyloxyethyl (2S,5R,6R)-6-[(2R)-2-amino- Internal standard solution—A solution of anhydrous
2-phenylacetylamino]-3,3-dimethyl-7-oxo-4-thia-1- caffeine in the mobile phase (1 in 25,000).
azabicyclo[3.2.0]heptane-2-carboxylate monohydrochloride Operating conditions—
[37661-08-8] Detector: An ultraviolet absorption photometer (wave-
length: 230 nm).
Bacampicillin Hydrochloride is a hydrochloride of Column: A stainless steel column 4.6 mm in inside diame-
ampicilline ethoxycarbonyloxyethyl ester. ter and 15 cm in length, packed with octadecylsilanized silica
It contains not less than 626 mg (potency) and not gel for liquid chromatography (5 mm in particle diameter).
more than 710 mg (potency) per mg, calculated on the Column temperature: A constant temperature of about
anhydrous basis. The potency of Bacampicillin Hydro- 259C.
chloride is expressed as mass (potency) of ampicillin Mobile phase: Dissolve 1.22 g of potassium dihydrogen
(C16H19N3O4S: 349.40). phosphate in water to make 900 mL, and add 100 mL of
acetonitrile.
Description Bacampicillin Hydrochloride occurs as a white Flow rate: Adjust so that the retention time of ampicillin
to pale yellow crystalline powder. is about 7 minutes.
It is freely soluble in methanol and in ethanol (95), and System suitability—
soluble in water. System performance: When the procedure is run with 10
Identification (1) Determine the absorption spectrum of a mL of the standard solution under the above operating con-
solution of Bacampicillin Hydrochloride in methanol (1 in ditions, ampicillin and the internal standard are eluted in this
1000) as directed under Ultraviolet-visible Spectrophotome- order with the resolution between these peaks being not less
try <2.24>, and compare the spectrum with the Reference than 5.
Spectrum or the spectrum of a solution of Bacampicillin Hy- System repeatability: When the test is repeated 6 times
drochloride RS prepared in the same manner as the sample with 10 mL of the standard solution under the above operat-
solution: both spectra exhibit similar intensities of absorp- ing conditions, the relative standard deviation of the ratio of
tion at the same wavelengths. the peak area of ampicillin to that of the internal standard is
(2) Determine the infrared absorption spectrum of not more than 2.0z.
JP XVII Official Monographs / Bacitracin 469
Water <2.48> Not more than 1.0z (0.5 g, volumetric titra- Identification (1) To 3 mL of a solution of Bacitracin
tion, direct titration). (1 in 100) add 3 mL of 4-dimethylaminobenzaldehyde TS,
shake until red-rosy to red-purple color appears, then add
several drops of a solution of sodium nitrite (1 in 100), and
Assay Weigh accurately an amount of Bacampicillin shake: a green to dark green color is produced.
Hydrochloride and Bacampicillin Hydrochloride RS, equiva- (2) Dissolve 60 mg each of Bacitracin and Bacitracin RS
lent to about 40 mg (potency), dissolve each in water to in 10 mL of water, and use these solutions as the sample so-
make exactly 100 mL, and use these solutions as the sample lution and standard solution. Perform the test with these so-
solution and standard solution. Perform the test with exactly lutions as directed under Thin-layer Chromatography <2.03>.
20 mL each of the sample solution and standard solution as Spot 1 mL each of the sample solution and standard solution
directed under Liquid Chromatography <2.01> according to on a plate of silica gel for thin-layer chromatography. De-
the following conditions, and determine the peak areas, AT velop the plate with a mixture of 1-butanol, acetic acid (100),
and AS, of bacampicillin in each solution. water, pyridine and ethanol (99.5) (30:15:10:6:5) to a dis-
tance of about 10 cm, and air-dry the plate. Spray evenly
Amount [ mg (potency)] of ampicillin (C16H19N3O4S)
ninhydrin TS on the plate, and heat at 1109C for 5 minutes:
＝ MS × AT/AS × 1000
the spots obtained from the sample solution and standard
MS: Amount [mg (potency)] of Bacampicillin Hydrochlo- solution show the same R f value.
ride RS taken
Operating conditions— Bacitracin according to Method 2, and perform the test.
Detector: An ultraviolet absorption photometer (wave- Prepare the control solution with 2.0 mL of Standard Lead
length: 254 nm). Solution (not more than 20 ppm).
Column: A stainless steel column 4.6 mm in inside diame- (2) Related substances—Dissolve 0.15 g of Bacitracin in
ter and 15 cm in length, packed with octadecylsilanized silica 0.05 mol/L sulfuric acid TS to make 100 mL. To 2 mL of
gel for liquid chromatography (5 mm in particle diameter). this solution add 0.05 mol/L sulfuric acid TS to make 10
Column temperature: A constant temperature of about mL, and determine the absorbances of this solution, A1 and
259 C. A2, at 252 nm and 290 nm as directed under Ultraviolet-
Mobile phase: To 500 mL of diluted 2 mol/L sodium dihy- visible Spectrophotometry <2.24>: A2/A1 is not more than
drogen phosphate TS (1 in 100), add diluted 0.05 mol/L dis- 0.20.
odium hydrogen phosphate TS (2 in 5) to adjust the pH to
6.8. To 500 mL of this solution add 500 mL of acetonitrile.
um, 609C, 3 hours).
Flow rate: Adjust so that the retention time of bacampicil-
lin is about 6.5 minutes. Residue on ignition <2.44> Not more than 1.0z (1 g).
System performance: When the procedure is run with
20 mL of the standard solution under the above operating
conditions, the number of theoretical plates and the symme-
(i) Test organism—Micrococcus luteus ATCC 10240.
try factor of the peak of bacampicillin are not less than
(ii) Culture medium—Use the medium iii in 3) under (1)
10,000 and not more than 2, respectively.
Agar media for seed and base layer.
(iii) Standard solutions—Weigh accurately an amount of
Bacitracin RS, equivalent to about 400 units, dissolve in
ing conditions, the relative standard deviation of peak areas
phosphate buffer solution (pH 6.0) to make exactly 20 mL,
of bacampicillin is not more than 2.0z.
and use this solution as the standard stock solution. Keep the
Containers and storage Containers—Tight containers. standard stock solution at not exceeding 109C and use within
2 days. Take exactly a suitable amount of the standard stock
solution before use, add phosphate buffer solution (pH 6.0)
Bacitracin to make solutions so that each mL contains 2 units and 0.5
units, and use these solutions as the high concentration
バシトラシン standard solution and low concentration standard solution,
respectively.
[1405-87-4] (iv) Sample solutions—Weigh accurately an amount of
Bacitracin, equivalent about 400 units, dissolve in phosphate
Bacitracin is a mixture of peptide substances having buffer solution (pH 6.0) to make exactly 20 mL. Take ex-
antibacterial activity including bacitracin A as the actly a suitable amount of this solution, add phosphate
main component produced by the growth of Bacillus buffer solution (pH 6.0) to make solutions so that each mL
subtilis or Bacillus licheniformis. contains 2 units and 0.5 units, and use these solutions as the
It contains not less than 60 Units per mg, calculated high concentration sample solution and low concentration
on the dried basis. The potency of Bacitracin is sample solution, respectively.
expressed as unit calculated from the amount of
bacitracin A (C66H103N17O16S: 1422.69). One unit of
Storage—In a cold place.
Bacitracin is equivalent to 23.8 mg of bacitracin A
(C66H103N17O16S).
Description Bacitracin occurs as a white to light brown
powder.
It is freely soluble in water, and slightly soluble in ethanol
(99.5).
470 Baclofen / Official Monographs JP XVII
length: 268 nm).
Baclofen Column: A stainless steel column 4 mm in inside diameter
バクロフェン for liquid chromatography (10 mm in particle diameter).
259C.
Mobile phase: A mixture of methanol and diluted acetic
acid (100) (1 in 900) (3:2).
Flow rate: Adjust so that the retention time of baclofen is
about 4 minutes.
C10H12ClNO2: 213.66 Time span of measurement: About 3 times as long as the
(3RS )-4-Amino-3-(4-chlorophenyl)butanoic acid retention time of baclofen, beginning after the solvent peak.
[1134-47-0] System suitability—
Test for required detectability: Adjust the sensitivity so
Baclofen contains not less than 98.5z of baclofen that the peak height of baclofen obtained from 25 mL of the
(C10H12ClNO2), calculated on the anhydrous basis. standard solution (1) is between 5 and 10 mm.
System performance: Dissolve 0.40 g of Baclofen and 5
Description Baclofen occurs as a white to pale yellowish
mg of methyl parahydroxybenzoate in 200 mL of the mobile
phase. To 10 mL of this solution add the mobile phase to
It is freely soluble in acetic acid (100), slightly soluble in
make 100 mL. When the procedure is run with 25 mL of this
water, very slightly soluble in methanol and in ethanol (95),
solution under the above operating conditions, baclofen and
and practically insoluble in diethyl ether.
methyl parahydroxybenzoate are eluted in this order with the
Identification (1) To 5 mL of a solution of Baclofen (1 in System repeatability: When the test is repeated 6 times
1000) add 1 mL of ninhydrin TS, and heat on a water bath with 25 mL of the standard solution (1) under the above
for 3 minutes: a blue-purple color develops. operating conditions, the relative standard deviation of the
(2) Determine the absorption spectrum of a solution of peak heights of baclofen is not more than 3.0z.
Baclofen in 0.1 mol/L hydrochloric acid TS (1 in 2000) as
Water <2.48> Not more than 1.0z (1 g, direct titration).
and compare the spectrum with the Reference Spectrum or Residue on ignition <2.44> Not more than 0.3z (1 g).
the spectrum of a solution of Baclofen RS prepared in the
Assay Weigh accurately about 0.5 g of Baclofen, dissolve
same manner as the sample solution: both spectra exhibit
in 80 mL of acetic acid (100), and titrate <2.50> with 0.1
mol/L perchloric acid VS until the color of the solution
(3) Perform the test with Baclofen as directed under
changes from purple through blue to greenish blue (indica-
tor: 2 drops of crystal violet TS). Perform a blank determi-
Purity (1) Chloride <1.03>—Dissolve 0.5 g of Baclofen in nation, and make any necessary correction.
50 mL of acetic acid (100), and add water to make 100 mL.
To 10 mL of this solution add 6 mL of dilute nitric acid and
the test solution. Prepare the control solution as follows: to Containers and storage Containers—Well-closed contain-
0.30 mL of 0.01 mol/L hydrochloric acid VS add 5 mL of ers.
acetic acid (100), 6 mL of dilute nitric acid and water to
(2) Heavy metals <1.07>—Proceed with 2.0 g of Baclofen Baclofen Tablets
according to Method 2, and perform the test. Prepare the
control solution with 2.0 mL of Standard Lead Solution (not バクロフェン錠
more than 10 ppm).
Baclofen Tablets contain not less than 93.0z and
of Baclofen according to Method 3, and perform the test
baclofen (C10H12ClNO2: 213.66).
(4) Related substances—Dissolve 50 mg of Baclofen in
50 mL of the mobile phase, and use this solution as the sam- Method of preparation Prepare as directed under Tablets,
ple solution. Pipet 1.0 mL and 1.5 mL of the sample solu- with Baclofen.
tion, to each add the mobile phase to make exactly 100 mL,
Identification (1) To a portion of powdered Baclofen
and use these solutions as the standard solutions (1) and (2),
Tablets, equivalent to 0.01 g of Baclofen, add 10 mL of
respectively. Perform the test with exactly 25 mL each of the
water, shake well, and filter. To 5 mL of the filtrate add 1
sample solution and standard solutions (1) and (2) as di-
mL of ninhydrin TS, and proceed as directed in the Identifi-
rected under Liquid Chromatography <2.01> according to
cation (1) under Baclofen.
the following conditions. Determine each peak height of
(2) To a portion of powdered Baclofen Tablets, equiva-
these solutions: each height of the peaks other than the peak
lent to 25 mg of Baclofen, add 50 mL of 0.1 mol/L hydro-
of baclofen from the sample solution is not larger than the
chloric acid TS, shake for 15 minutes, and filter. Determine
peak height of baclofen from the standard solution (1), and
the absorption spectrum of the filtrate as directed under Ul-
the total height of these peaks is not larger than the peak
traviolet-visible Spectrophotometry <2.24>: it exhibits maxi-
height of baclofen from the standard solution (2).
ma between 257 nm and 261 nm, between 264 nm and 268
nm, and between 272 nm and 276 nm.
(3) To a portion of powdered Baclofen Tablets, equiva-
JP XVII Official Monographs / Bamethan Sulfate 471
lent to 0.01 g of Baclofen, add 2 mL of a mixture of metha- under Ultraviolet-visible Spectrophotometry <2.24>.
nol and acetic acid (100) (4:1), shake well, centrifuge, and
of baclofen (C10H12ClNO2)
dissolve 0.01 g of Baclofen RS in 2 mL of a mixture of meth-
＝ MS × AT/AS × V?/V × 1/C × 50
anol and acetic acid (100) (4:1), and use this solution as the
standard solution. Perform the test with these solutions as MS: Amount (mg) of Baclofen RS taken, calculated on the
directed under Thin-layer Chromatography <2.03>. Spot 20 anhydrous basis
mL each of the sample solution and standard solution on a C: Labeled amount (mg) of baclofen (C10H12ClNO2) in 1
plate of silica gel with fluorescent indicator for thin-layer tablet
chromatography. Develop the plate with a mixture of 1-
butanol, water and acetic acid (100) (4:1:1) to a distance of
Baclofen Tablets. Weigh accurately a portion of the powder,
equivalent to about 50 mg of baclofen (C10H12ClNO2), add
let light (main wavelength: 254 nm): the spot from the sam-
130 mL of 0.1 mol/L hydrochloric acid TS, shake for 10
ple solution and that from the standard solution show the
minutes, add 0.1 mol/L hydrochloric acid TS to make ex-
same R f value.
actly 200 mL, and centrifuge. Pipet 10 mL of the superna-
Uniformity of dosage units <6.02> Perform the test accord- tant liquid, add 2 drops of phenolphthalein TS, neutralize
ing to the following method: it meets the requirement of the with dilute sodium hydroxide TS, add water to make exactly
Content uniformity test. 50 mL, and use this solution as the sample solution. Sepa-
To 1 tablet of Baclofen Tablets add 5 mL of 0.1 mol/L hy- rately, weigh accurately about 0.25 g of Baclofen RS
drochloric acid TS, disperse the tablet into small particles (separately determine the water content <2.48> in the same
with the aid of ultrasonic waves, then shake for 10 minutes, manner as Baclofen), and dissolve in 0.1 mol/L hydrochloric
and add 0.1 mol/L hydrochloric acid TS to make exactly acid TS to make exactly 100 mL. Pipet 10 mL of this solu-
V mL so that each mL contains about 0.5 mg of baclofen tion, and add 0.1 mol/L hydrochloric acid TS to make
(C10H12ClNO2). Centrifuge, pipet 5 mL of the supernatant exactly 100 mL. Pipet 10 mL of this solution, add 2 drops of
liquid, add 2 drops of phenolphthalein TS, neutralize with phenolphthalein TS, neutralize with dilute sodium hydroxide
dilute sodium hydroxide TS, then add water to make exactly TS, add water to make exactly 50 mL, and use this solution
50 mL, and use this solution as the sample solution. Sepa- as the standard solution. Pipet 2 mL each of the sample solu-
rately, weigh accurately about 25 mg of Baclofen RS tion and the standard solution, to each add 4 mL of nin-
(separately determine the water <2.48> in the same manner as hydrin-stannous chloride TS, shake, heat on a water bath for
Baclofen), and dissolve in 0.1 mol/L hydrochloric acid TS to 20 minutes, and shake at once vigorously for 2 minutes.
make exactly 50 mL. Pipet 5 mL of this solution, add 2 After cooling, to each solution add a mixture of water and 1-
drops of phenolphthalein TS, neutralize with dilute sodium propanol (1:1) to make exactly 25 mL. Determine the absor-
hydroxide TS, then add water to make exactly 50 mL, and bances, AT and AS, of these solutions at 570 nm as directed
use this solution as the standard solution. To exactly 2 mL under Ultraviolet-visible Spectrophotometry <2.24>, using a
each of the sample solution and standard solution add 4 mL blank prepared with 2 mL of water in the same manner.
of ninhydrin-tin (II) chloride TS, mix, heat on a water bath
Amount (mg) of baclofen (C10H12ClNO2)
for 20 minutes, then immediately shake vigorously for
＝ MS × AT/AS × 1/5
2 minutes. After cooling, add a mixture of water and 1-
propanol (1:1) to make them exactly 25 mL, and determine MS: Amount (mg) of Baclofen RS taken, calculated on the
the absorbances, AT and AS, of them at 570 nm as directed anhydrous basis
under Ultraviolet-visible Spectrophotometry <2.24>, using a
solution obtained with 2 mL of water by the same procedure
ers.
as above as the blank.
Amount (mg) of baclofen (C10H12ClNO2)
＝ MS × AT/AS × V/50 Bamethan Sulfate
MS: Amount (mg) of Baclofen RS taken, calculated on the
バメタン硫酸塩
anhydrous basis
in 45 minutes of Baclofen Tablets is not less than 70z.
Start the test with 1 tablet of Baclofen Tablets, withdraw
not less than 20 mL of the medium at the specified minute (C12H19NO2)2.H2SO4: 516.65
after starting the test, and filter through a membrane filter (1RS )-2-Butylamino-1-
with a pore size not exceeding 0.8 mm. Discard the first 10 (4-hydroxyphenyl)ethanol hemisulfate
mL of the filtrate, pipet V mL of the subsequent, add water [5716-20-1]
to make exactly V? mL so that each mL contains about 10 mg
of baclofen (C10H12ClNO2), and use this solution as the sam- Bamethan Sulfate, when dried, contains not less
ple solution. Separately, weigh accurately about 10 mg of than 99.0z of bamethan sulfate [(C12H19NO2)2.
Baclofen RS (separately determine the water <2.48> in the H2SO4].
same manner as Baclofen), dissolve in water to make exactly
Description Bamethan Sulfate occurs as white, crystals or
100 mL, then pipet 10 mL of this solution, add water to
crystalline powder. It is odorless, and has a bitter taste.
It is freely soluble in water and in acetic acid (100), soluble
solution. Determine the absorbances, AT and AS, of the sam-
in methanol, slightly soluble in ethanol (95), and practically
ple solution and the standard solution at 220 nm as directed
472 Barbital / Official Monographs JP XVII
insoluble in diethyl ether. Each mL of 0.1 mol/L perchloric acid VS
Melting point: about 1699C (with decomposition). ＝ 51.67 mg of (C12H19NO2)2.H2SO4
Identification (1) To 1 mL of a solution of Bamethan Containers and storage Containers—Tight containers.
Sulfate (1 in 1000) add 5 mL of a solution of 4-nitroben-
zenediazonium fluoroborate (1 in 2000) and 10 mL of boric
acid-potassium chloride-sodium hydroxide buffer solution Barbital
(pH 9.2): an orange-red color develops.
(2) Determine the absorption spectrum of a solution of バルビタール
Bamethan Sulfate in 0.01 mol/L hydrochloric acid TS (1 in
C8H12N2O3: 184.19
Bamethan Sulfate, previously dried, as directed in the potas-
5,5-Diethylpyrimidine-2,4,6(1H,3H,5H )-trione
[57-44-3]
try <2.25>: it exhabits absorption at the wave numbers of
about 1618, 1597, 1518, 1118 and 833 cm－1.
Barbital, when dried, contains not less than 99.0z
(4) A solution of Bamethan Sulfate (1 in 100) responds
of barbital (C8H12N2O3).
to the Qualitative Tests <1.09> for sulfate.
Description Barbital occurs as colorless or white crystals or
pH <2.54> Dissolve 1.0 g of Bamethan Sulfate in 10 mL of
a white crystalline powder. It is odorless, and has a slightly
bitter taste.
Purity (1) Clarity and color of solution—Dissolve 1.0 g It is freely soluble in acetone and in pyridine, soluble in
of Bamethan Sulfate in 20 mL of water: the solution is clear, ethanol (95), sparingly soluble in diethyl ether, and slightly
and has no more color than the following control solution. soluble in water and in chloroform.
Control solution: To 1.5 mL of Matching Fluid O add It dissolves in sodium hydroxide TS and in ammonia TS.
diluted hydrochloric acid (1 in 40) to make 200 mL. The pH of its saturated solution is between 5.0 and 6.0.
(2) Chloride <1.03>—Perform the test with 3.5 g of
Identification (1) Boil 0.2 g of Barbital with 10 mL of
Bamethan Sulfate. Prepare the control solution with 0.25
sodium hydroxide TS: the gas evolved changes moistened red
mL of 0.01 mol/L hydrochloric acid VS (not more than
litmus paper to blue.
0.002z).
(2) Dissolve 0.05 g of Barbital in 5 mL of diluted pyri-
(3) Heavy metals <1.07>—Proceed with 2.0 g of Bame-
dine (1 in 10), add 0.3 mL of copper (II) sulfate TS, shake,
than Sulfate according to Method 1, and perform the test.
and allow to stand for 5 minutes: a red-purple precipitate is
formed. Shake the mixture with 5 mL of chloroform: a red-
purple color develops in the chloroform layer. Separately,
dissolve 0.05 g of Barbital in 2 to 3 drops of ammonia-
of Bamethan Sulfate according to Method 3, and perform
ammonium chloride buffer solution (pH 10.7) and 5 mL of
diluted pyridine (1 in 10). Add 5 mL of chloroform and 0.3
(5) Related substances—Dissolve 0.10 g of Bamethan
mL of copper (II) sulfate TS to the solution: a red-purple
Sulfate in 2 mL of methanol, and use this solution as the
precipitate is produced in the aqueous layer. The red-purple
precipitate is not dissolved in the chloroform by shaking.
methanol to make exactly 100 mL, and use this solution as
(3) To 0.4 g of Barbital add 0.1 g of anhydrous sodium
the standard solution. Perform the test with these solutions
carbonate and 4 mL of water, shake, and add a solution of
as directed under Thin-layer Chromatography <2.03>. Spot 2
0.3 g of 4-nitrobenzyl chloride in 7 mL of ethanol (95). Heat
the mixture on a water bath under a reflux condenser for 30
plate of silica gel for thin-layer chromatography. Develop
minutes, and allow to stand for 1 hour. Collect the separated
the plate with a mixture of chloroform and methanol (7:2) in
crystals, wash with 7 mL of sodium hydroxide TS and a
a developing vessel saturated with ammonia vapor to a dis-
small amount of water, recrystallize from a mixture of
tance of about 12 cm, and air-dry the plate. Spray evenly
ethanol (95) and chloroform (1:1), and dry at 1059C for 30
Dragendorff's TS for spraying on the plate, air-dry for 15
minutes: the crystals melt <2.60> between 1929C and 1969C.
minutes, spray Dragendorff's TS for spraying again, then,
after 1 minute, spray evenly a solution of sodium nitrite (1 in Melting point <2.60> 189 – 1929
C
20), and immediately put a glass plate on the plate. Examine
the plate after 30 minutes: the spots other than the principal
of Barbital in 5 mL of sodium hydroxide TS: the solution is
spot from the sample solution are not more intense than the
spot from the standard solution.
(2) Chloride <1.03>—Dissolve 0.30 g of Barbital in 20
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, mL of acetone, and add 6 mL of dilute nitric acid and water
4 hours). to make 50 mL. Perform the test using this solution as the
test solution. Prepare the control solution as follows: take
0.30 mL of 0.01 mol/L hydrochloric acid VS, 20 mL of ace-
Assay Weigh accurately about 0.75 g of Bamethan Sulfate, tone and 6 mL of dilute nitric acid, and add water to make
previously dried, dissolve in 80 mL of acetic acid (100), and 50 mL (not more than 0.035z).
titrate with 0.1 mol/L perchloric acid VS (potentiometric (3) Sulfate <1.14>—Dissolve 0.40 g of Barbital in 20 mL
titration). Perform a blank determination, and make any of acetone, and add 1 mL of dilute hydrochloric acid and
necessary correction. water to make 50 mL. Perform the test using this solution as
JP XVII Official Monographs / Beclometasone Dipropionate 473
the test solution. Prepare the control solution as follows: (4) Heavy metals <1.07>—Boil 5.0 g of Barium Sulfate
take 0.40 mL of 0.005 mol/L sulfuric acid VS, 20 mL of ace- with 2.5 mL of acetic acid (100) and 50 mL of water for 10
tone, and 1 mL of dilute hydrochloric acid, and add water to minutes, cool, add 0.5 mL of ammonia TS and water to
make 50 mL (not more than 0.048z). make 100 mL, and filter. Perform the test with a 50-mL por-
(4) Heavy metals <1.07>—Proceed with 1.0 g of Barbital tion of this filtrate. Prepare the control solution with 2.5 mL
according to Method 2, and perform the test. Prepare the of Standard Lead Solution, 1.25 mL of acetic acid (100),
control solution with 2.0 mL of Standard Lead solution (not 0.25 mL of ammonia TS and water to make 50 mL (not
more than 20 ppm). more than 10 ppm).
(5) Readily carbonizable substances <1.15>—Perform the (5) Arsenic <1.11>—Prepare the test solution with 2.0 g
test with 0.5 g of Barbital. The solution is not more colored of Barium Sulfate according to Method 1, and perform the
than Matching Fluid A. test (not more than 1 ppm).
(6) Hydrochloric acid-soluble substances and soluble
barium salts—Cool the solution obtained in (3), add water to
2 hours).
make 100 mL, and filter. Evaporate 50 mL of the filtrate on
Residue on ignition <2.44> Not more than 0.1z (1 g). a water bath to dryness, add 2 drops of hydrochloric acid
and 10 mL of warm water, filter through filter paper for
Assay Weigh accurately about 0.4 g of Barbital, previously
assay, and wash with 10 mL of warm water. Evaporate the
dried, and dissolve in 5 mL of ethanol (95) and 50 mL of
combined filtrate and washings on a water bath to dryness,
chloroform. Titrate <2.50> with 0.1 mol/L potassium hy-
and dry the residue at 1059 C for 1 hour: the residue weighs
droxide-ethanol VS until the color of the solution changes
not more than 15 mg. Shake the residue, if any, with 10 mL
from yellow through light blue to purple (indicator: 1 mL of
of water, and filter. To the filtrate add 0.5 mL of dilute sul-
alizarin yellow GG-thymolphthalein TS). Perform a blank
furic acid, and allow to stand for 30 minutes: no turbidity is
produced.
Each mL of 0.1 mol/L potassium hydroxide-ethanol VS
＝ 18.42 mg of C8H12N2O3
ers.
ers.
Freeze-dried BCG Vaccine
Barium Sulfate (for Percutaneous Use)
乾燥 BCG ワクチン
硫酸バリウム
Freeze-dried BCG Vaccine (for Percutaneous Use) is

BaSO4: 233.39
a preparation for injection which is dissolved before
use.
Description Barium Sulfate occurs as a white powder. It is
It contains live bacteria derived from a culture of
odorless and tasteless.
the bacillus of Calmette and Gu áerin.
It conforms to the requirements of Freeze-dried
diethyl ether.
BCG Vaccine (for Percutaneous Use) in the Minimum
It does not dissolve in hydrochloric acid, in nitric acid and
Requirements for Biological Products.
in sodium hydroxide TS.
Description Freeze-dried BCG Vaccine (for Percutaneous
Identification (1) Mix 0.5 g of Barium Sulfate with 2 g
Use) becomes a white to light yellow, turbid liquid on addi-
each of anhydrous sodium carbonate and potassium carbon-
tion of solvent.
ate in a crucible, heat the mixture until fusion is complete,
treat the cooled mass with hot water, and filter. The filtrate,
acidified with hydrochloric acid, responds to the Qualitative
Tests <1.09> for sulfate. Beclometasone Dipropionate
(2) Wash the hot water-insoluble residue obtained in (1)
ベクロメタゾンプロピオン酸エステル
with water, dissolve in 2 mL of acetic acid (31), and filter, if
necessary: the solution responds to the Qualitative Tests
<1.09> for barium salt.
Purity (1) Acidity or alkalinity—Agitate 1.0 g of Barium
Sulfate with 20 mL of water for 5 minutes: the solution is
neutral.
(2) Phosphate—Boil 1.0 g of Barium Sulfate with 3 mL
of nitric acid and 5 mL of water for 5 minutes, cool, and add
water to restore the original volume. Filter through a filter
paper, previously washed with dilute nitric acid, to the fil-
C28H37ClO7: 521.04
trate add an equal volume of hexaammonium heptamolyb-
9-Chloro-11b,17,21-trihydroxy-16b-methylpregna-1,4-
date TS, and allow to stand between 509 C and 609 C for 1
diene-3,20-dione 17,21-dipropanoate
hour: no yellow precipitate is produced.
[5534-09-8]
(3) Sulfide—Place 10 g of Barium Sulfate in a 250-mL
conical flask, add 10 mL of dilute hydrochloric acid and
Beclometasone Dipropionate, when dried, contains
water to make 100 mL, and boil for 10 minutes: the gas
evolved does not darken moistened lead (II) acetate paper.
474 Bekanamycin Sulfate / Official Monographs JP XVII
beclometasone dipropionate (C28H37ClO7). lution and the standard solution, respectively. Perform the
Description Beclometasone Dipropionate occurs as a white
to pale yellow powder.
It is soluble in methanol, sparingly soluble in ethanol (95)
QT and QS, of the peak area of beclometasone dipropionate
and in 1,4-dioxane, and practically insoluble in water.
to that of the internal standard, respectively.
It shows crystal polymorphism. Amount (mg) of beclometasone dipropionate (C28H37ClO7)
＝ M S × QT / QS
Identification (1) Dissolve 2 mg of Beclometasone
Dipropionate in 2 mL of sulfuric acid: initially a yellowish MS: Amount (mg) of Beclometasone Dipropionate RS
color develops, and gradually changes through orange to taken
dark red-brown. To this solution add carefully 10 mL of
Internal standard solution—A solution of testosterone
water: the color changes to bluish green, and a flocculent
propionate in methanol (1 in 4000).
precipitate is formed.
(2) Dissolve 0.01 g of Beclometasone Dipropionate in 1
mL of methanol, add 1 mL of Fehling's TS, and heat: a red
length: 254 nm).
to red-brown precipitate is formed.
(3) Perform the test with 0.02 g of Beclometasone
Dipropionate as directed under Oxygen Flask Combustion
Method <1.06>, using a mixture of 1 mL of sodium hydrox-
ide TS and 20 mL of water as an absorbing liquid: the test
259C.
solution responds to the Qualitative Tests <1.09> for chlo-
Mobile phase: A mixture of acetonitrile and water (3:2).
ride.
Flow rate: Adjust so that the retention time of beclometa-
sone dipropionate is about 6 minutes.
Beclometasone Dipropionate, previously dried, as directed in
erence Spectrum or the spectrum of previously dried Beclo-
ditions, beclometasone dipropionate and the internal stand-
metasone Dipropionate RS: both spectra exhibit similar in-
ard are eluted in this order with the resolution between these
tensities of absorption at the same wave numbers. If any
peaks being not less than 8.
difference appears between the spectra, dissolve Beclometa-
sone Dipropionate and Beclometasone Dipropionate RS in
ethanol (95), respectively, then evaporate the ethanol to dry-
ness, and repeat the test on the residues.
of the peak area of beclometasone dipropionate to that of
D : ＋88 – ＋949 (after drying, the internal standard is not more than 1.0z.
0.1 g, 1,4-dioxane, 10 mL, 100 mm).
Beclometasone Dipropionate according to Method 2, and
perform the test. Prepare the control solution with 1.5 mL of Bekanamycin Sulfate
(2) Related substances—Dissolve 20 mg of Beclometa- ベカナマイシン硫酸塩
sone Dipropionate in 5 mL of a mixture of chloroform and
methanol (9:1), and use this solution as the sample solution.
Pipet 1 mL of the sample solution, add a mixture of chlo-
roform and methanol (9:1) to make exactly 50 mL, and use
phy <2.03>. Spot 5 mL each of the sample solution and
standard solution on a plate of silica gel for thin-layer
chromatography. Develop the plate with a mixture of 1,2-
dichloroethane, methanol and water (475:25:1) to a distance
of about 15 cm, and air-dry the plate. Spray evenly alkaline
blue tetrazolium TS on the plate: the spots other than the
principal spot from the sample solution are not more intense
Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, C18H37N5O10.xH2SO4
3 hours). 3-Amino-3-deoxy-a-D-glucopyranosyl-(1→6)-[2,6-
diamino-2,6-dideoxy-a-D-glucopyranosyl-(1→4)]-2-deoxy-
D-streptamine sulfate
Assay Weigh accurately about 20 mg each of Beclometa- [70550-99-1]
sone Dipropionate and Beclometasone Dipropionate RS,
previously dried, and dissolve each in methanol to make Bekanamycin Sulfate is the sulfate of an
exactly 50 mL. Pipet 10 mL each of these solutions, add aminoglycoside substance having antibacterial activity
exactly 10 mL of the internal standard solution and metha- produced by the growth of the mutant of Strep-
nol to make 50 mL, and use these solutions as the sample so- tomyces kanamyceticus.
JP XVII Official Monographs / Benidipine Hydrochloride 475
It contains not less than 680 mg (potency) and not (i) Test organism—Bacillus subtilis ATCC 6633
more than 770 mg (potency) per mg, calculated on (ii) Culture medium—Use the medium i in 1) under (1)
the dried basis. The potency of Bekanamycin Sulfate Agar media for seed and base layer having pH <2.54> 7.8 to
is expressed as mass (potency) of bekanamycin 8.0 after sterilization.
(C18H37N5O10: 483.51). (iii) Standard solutions—Weigh accurately an amount of
Bekanamycin Sulfate RS, previously dried, equivalent to
Description Bekanamycin Sulfate occurs as a white pow-
about 20 mg (potency), dissolve in diluted phosphate buffer
der.
solution (pH 6.0) (1 in 2) to make exactly 50 mL, and use
this solution as the standard stock solution. Keep the stand-
ethanol (99.5).
ard stock solution at 5 to 159C and use within 30 days. Take
Identification (1) Dissolve 20 mg of Bekanamycin Sulfate exactly a suitable amount of the standard stock solution
in 2 mL of 1/15 mol/L phosphate buffer solution (pH 5.6), before use, add 0.1 mol/L phosphate buffer solution (pH
add 1 mL of ninhydrin TS, and boil: a blue-purple color 8.0) to make solutions so that each mL contains 10 mg (po-
develops. tency) and 2.5 mg (potency), and use these solutions as the
(2) Dissolve 30 mg each of Bekanamycin Sulfate and high concentration standard solution and low concentration
Bekanamycin Sulfate RS in 5 mL of water, and use these standard solution, respectively.
solutions as the sample solution and standard solution. (iv) Sample solutions—Weigh accurately an amount of
Perform the test with these solutions as directed under Thin- Bekanamycin Sulfate, equivalent to about 20 mg (potency),
layer Chromatography <2.03>. Spot 5 mL each of the sample and dissolve in water to make exactly 50 mL. Take exactly a
solution and standard solution on a plate of silica gel for suitable amount of this solution, add 0.1 mol/L phosphate
thin-layer chromatography. Develop the plate with a solu- buffer solution (pH 8.0) to make solutions so that each mL
tion of potassium dihydrogen phosphate (3 in 40) to a dis- contains 10 mg (potency) and 2.5 mg (potency), and use these
tance of about 10 cm, and air-dry the plate. Spray evenly solutions as the high concentration sample solution and low
0.2z ninhydrin-water saturated 1-butanol TS, and heat at concentration sample solution, respectively.
1009C for 10 minutes: the principal spots obtained from the
sample solution and the standard solution show a purple-
brown color and the same R f value.
(3) To a solution of Bekanamycin Sulfate (1 in 5) add 1
drop of barium chloride TS: a white turbidity is produced. Benidipine Hydrochloride
D : ＋102 – ＋1169(after drying, ベニジピン塩酸塩
0.25 g, water, 25 mL, 100 mm).
0.50 g of Bekanamycin Sulfate in 10 mL of water is between
6.0 and 8.5.
of Bekanamycin Sulfate in 5 mL of water: the solution is
(2) Heavy metals <1.07>—Proceed with 1.0 g of Bekana-
mycin Sulfate according to Method 4, and perform the test. C28H31N3O6.HCl: 542.02
Prepare the control solution with 3.0 mL of Standard Lead 3-[(3RS )-1-Benzylpiperidin-3-yl] 5-methyl (4RS )-
Solution (not more than 30 ppm). 2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydropyridine-3,5-
(3) Arsenic <1.11>—Prepare the test solution with 2.0 g dicarboxylate monohydrochloride
of Bekanamycin Sulfate according to Method 1, and per- [91599-74-5]
(4) Related substances—Dissolve 60 mg of Bekanamycin Benidipine Hydrochloride, when dried, contains
Sulfate in 10 mL of water, and use this solution as the sam- not less than 99.0z and not more than 101.0z of
ple solution. Pipet 3 mL of the sample solution, add water to benidipine hydrochloride (C28H31N3O6.HCl).
Description Benidipine Hydrochloride occurs as a yellow
crystalline powder.
under Thin-layer Chromatography <2.03>. Spot 5 mL each of
It is very soluble in formic acid, soluble in methanol,
sparingly soluble in ethanol (99.5), and practically insoluble
gel for thin-layer chromatography. Develop the plate with a
in water.
solution of potassium dihydrogen phosphate (3 in 40) to a
A solution of Benidipine Hydrochloride in methanol (1 in
distance of about 10 cm, and air-dry the plate. Spray evenly
100) shows no optical rotation.
0.2z ninhydrin-water saturated 1-butanol TS on the plate,
and heat at 1009 C for 10 minutes: the spot other than the
principal spot obtained from the sample solution is not more Identification (1) Determine the absorption spectrum of a
intense than the spot obtained from the standard solution. solution of Benidipine Hydrochloride in methanol (1 in
Residue on ignition <2.44> Not more than 0.5z (1 g). sorption at the same wavelengths.
Benidipine Hydrochloride, previously dried, as directed in
476 Benidipine Hydrochloride Tablets / Official Monographs JP XVII
photometry <2.25>, and compare the spectrum with the Ref- Residue on ignition <2.44> Not more than 0.1z (1 g).
Assay Weigh accurately about 0.7 g of Benidipine Hydro-
chloride, previously dried, dissolve in 10 mL of formic acid,
(3) To 5 mL of a solution of Benidipine Hydrochloride
add 70 mL of acetic anhydride, and titrate <2.50> with 0.1
(1 in 10) add 5 mL of ammonia TS, heat on a water bath for
5 minutes, cool, and filter. The filtrate, which is acidified
form a blank determination in the same manner, and make
with dilute nitric acid, responds to the Qualitative Tests
<1.09> (2) for chloride.
＝ 54.20 mg of C28H31N3O6.HCl
Benidipine Hydrochloride according to Method 2, and per-
form the test. Prepare the control solution with 2.0 mL of Containers and storage Containers—Tight containers.
(2) Related substances—Dissolve 20 mg of Benidipine
Hydrochloride in 100 mL of a mixture of water and metha- Benidipine Hydrochloride Tablets
nol (1:1), and use this solution as the sample solution. Pipet
1 mL of the sample solution, add the mixture of water and ベニジピン塩酸塩錠
methanol (1:1) to make exactly 500 mL, and use this solution
Benidipine Hydrochloride Tablets contain not
the labeled amount of benidipine hydrochloride
(C28H31N3O6.HCl: 542.02).
automatic integration method: the peak areas of bisbenzyl-
piperidyl ester having the relative retention time of about Method of preparation Prepare as directed under Tablets,
0.35 to benidipine, dehydro derivative having the relative with Benidipine Hydrochloride.
retention time of about 0.75 and other related substances are
Identification Shake well a quantity of powdered Benidi-
not larger than 1/2 times the peak area of benidipine with
pine Hydrochloride Tablets, equivalent to 10 mg of Benidi-
the standard solution, and the total area of the peaks other
pine Hydrochloride, with 100 mL of methanol, and centri-
than benidipine is not larger than the peak area of benidipine
fuge. To 10 mL of the supernatant liquid add methanol to
with the standard solution. For the areas of the peaks of bis-
benzylpiperidyl ester and dehydro derivative, multiply their
Determine the absorption spectrum of the sample solution as
relative response factor 1.6, respectively.
directed under Ultraviolet-visible Spectrophotometry <2.24>:
it exhibits maxima between 235 nm and 239 nm, and between
350 nm and 360 nm.
length: 237 nm).
Column: A stainless steel column 4.6 mm in inside diame- Purity Dehydro derivative—Powder Benidipine Hydro-
ter and 10 cm in length, packed with octadecylsilanized silica chloride Tablets in an agate mortar. To an amount of the
gel for liquid chromatography (3 mm in particle diameter). powder, equivalent to 20 mg of Benidipine Hydrochloride,
Column temperature: A constant temperature of about add about 80 mL of a mixture of diluted phosphoric acid (1
259 C. in 500) and methanol (1:1), shake well, and add the mixture
Mobile phase: A mixture of 0.05 mol/L potassium dihy- of diluted phosphoric acid (1 in 500) and methanol (1:1) to
drogen phosphate TS (pH 3.0), methanol and tetrahydrofu- make exactly 100 mL. Filter through a membrane filter with
ran (65:27:8). pore size of 0.45 mm, and use the filtrate as the sample solu-
Flow rate: Adjust so that the retention time of benidipine tion. Separately, dissolve 20 mg of benidipine hydrochloride
is about 20 minutes. for assay in the mixture of diluted phosphoric acid (1 in 500)
Time span of measurement: About 2 times as long as the and methanol (1:1) to make exactly 100 mL. Pipet 1 mL of
retention time of benidipine, beginning after the solvent this solution, add the mixture of diluted phosphoric acid (1
peak. in 500) and methanol (1:1) to make exactly 100 mL, and use
System suitability— this solution as the standard solution. Perform the test with
Test for required detectability: Measure exactly 5 mL of exactly 10 mL each of the sample solution and standard
the standard solution, and add the mixture of water and solution as directed under Liquid Chromatography <2.01>
methanol (1:1) to make exactly 20 mL. Confirm that the according to the following conditions, and determine each
peak area of benidipine obtained with 10 mL of this solution peak area by the automatic integration method: the peak
is equivalent to 18 to 32z of that obtained with 10 mL of the area of dehydro derivative having the relative retention time
standard solution. of about 0.75 to benidipine is not larger than 1/2 times the
System performance: Dissolve 6 mg of Benidipine Hydro- peak area of benidipine with the standard solution. For the
chloride and 5 mg of benzoin in 200 mL of the mixture of area of the peak of dehydro derivative, multiply the relative
water and methanol (1:1). When the procedure is run with 10 response factor 1.6.
mL of this solution under the above operating conditions, Operating conditions—
benzoin and benidipine are eluted in this order with the reso- Perform as directed in the operating conditions in the
lution between these peaks being not less than 8. Assay.
System repeatability: When the test is repeated 6 times System suitability—
with 10 mL of the standard solution under the above operat- Test for required detectability: Measure exactly 2 mL of
ing conditions, the relative standard deviation of the peak the standard solution, and add the mixture of diluted phos-
area of benidipine is not more than 3.5z. phoric acid (1 in 500) and methanol (1:1) to make exactly 20
mL. Confirm that the peak area of benidipine obtained with
2 hours).
JP XVII Official Monographs / Benidipine Hydrochloride Tablets 477
tained with 10 mL of the standard solution. MS: Amount (mg) of benidipine hydrochloride for assay
System performance: Dissolve 6 mg of benidipine hydro- taken
chloride and 5 mg of benzoin in 200 mL of a mixture of C: Labeled amount (mg) of benidipine hydrochloride
water and methanol (1:1). When the procedure is run with 10 (C28H31N3O6.HCl) in 1 tablet.
mL of this solution under the above operating conditions,
benzoin and benidipine are eluted in this order with the reso-
lution between these peaks being not less than 8.
length: 237 nm).
area of benidipine is not more than 2.0z.
Uniformity of dosage units <6.02> Perform the test accord- 259C.
ing to the following method: it meets the requirement of the Mobile phase: A mixture of 0.05 mol/L potassium dihy-
Content uniformity test. drogen phosphate TS (pH 3.0) and acetonitrile (11:9).
To 1 tablet of Benidipine Hydrochloride Tablets add 40 Flow rate: Adjust so that the retention time of benidipine
mL of a mixture of diluted phosphoric acid (1 in 500) and is about 5 minutes.
methanol (1:1), shake to disintegrate, and add a suitable System suitability—
amount of the mixture of diluted phosphoric acid (1 in 500) System performance: When the procedure is run with 50
and methanol (1:1) to make exactly V mL of a solution, con- mL of the standard solution under the above operating con-
taining 40 mg of benidipine hydrochloride (C28H31N3O6.HCl) ditions, the number of theoretical plates and the symmetry
per mL. Centrifuge the solution, pipet 20 mL of the superna- factor of the peak of benidipine are not less than 3000 and
tant liquid, add exactly 10 mL of the internal standard solu- not more than 2.0, respectively.
tion and the mixture of diluted phosphoric acid (1 in 500) System repeatability: When the test is repeated 6 times
and methanol (1:1) to make 50 mL, and use this solution as with 50 mL of the standard solution under the above operat-
the sample solution. Then, proceed as directed in the Assay. ing conditions, the relative standard deviation of the peak
area of benidipine is not more than 1.5z.
Amount (mg) of benidipine hydrochloride
(C28H31N3O6.HCl) Assay Weigh accurately the mass of not less than 20
＝ MS × QT/QS × V/1000 Benidipine Hydrochloride Tablets, and powder using an
agate mortar. Weigh accurately a part of the powder,
MS: Amount (mg) of benidipine hydrochloride for assay
equivalent to about 8 mg of benidipine hydrochloride
taken
(C28H31N3O6.HCl), add about 150 mL of a mixture of
Internal standard solution—A solution of benzoin in a mix- diluted phosphoric acid (1 in 500) and methanol (1:1), shake,
ture of water and methanol (1:1) (13 in 200,000). then add the mixture of diluted phosphoric acid (1 in 500)
and methanol (1:1) to make exactly 200 mL, and centrifuge.
Pipet 20 mL of the supernatant liquid, add exactly 10 mL of
the internal standard solution and the mixture of diluted
sinker, using 900 mL of 1st fluid for dissolution test as the
phosphoric acid (1 in 500) and methanol (1:1) to make 50
dissolution medium, the dissolution rate of a 2-mg tablet and
a 4-mg tablet in 30 minutes is not less than 80z, and that of
weigh accurately about 40 mg of benidipine hydrochloride
a 8-mg tablet in 45 minutes is not less than 85z.
for assay, previously dried at 1059C for 2 hours, and dis-
Start the test with 1 tablet of Benidipine Hydrochloride
solve in the mixture of diluted phosphoric acid (1 in 500) and
methanol (1:1) to make exactly 100 mL. Pipet 2 mL of this
solution, add exactly 10 mL of the internal standard solution
and the mixture of diluted phosphoric acid (1 in 500) and
Discard the first 10 mL of the filtrate pipet the subsequent
methanol (1:1) to make 50 mL, and use this solution as the
V mL, and add the dissolution medium to make exactly
V? mL so that each mL contains about 2.2 mg of benidipine
hydrochloride (C28H31N3O6.HCl). Pipet 5 mL of this solu-
tion, add exactly 5 mL of the mobile phase, and use this so-
ditions, and calculate the ratios, QT and QS, of the peak area
of benidipine to that of the internal standard.
about 22 mg of benidipine hydrochloride for assay, previ-
ously dried at 1059 C for 2 hours, and dissolve in the mobile Amount (mg) of benidipine hydrochloride
phase to make exactly 100 mL. Pipet 2 mL of this solution, (C28H31N3O6.HCl)
and add the mobile phase to make exactly 50 mL. Pipet 5 ＝ MS × QT/QS × 1/5
mL of this solution, and add the mobile phase to make ex-
MS: Amount (mg) of benidipine hydrochloride for assay
taken
of the dissolution medium, and use this solution as the
standard solution. Perform the test with exactly 50 mL each Internal standard solution—A solution of benzoin in a mix-
of the sample solution and standard solution as directed ture of water and methanol (1:1) (13 in 200,000).
under Liquid Chromatography <2.01> according to the fol- Operating conditions—
lowing conditions, and determine the peak areas, AT and AS, Detector: An ultraviolet absorption photometer (wave-
of benidipine in each solution. length: 237 nm).
Dissolution rate (z) of benidipine hydrochloride
(C28H31N3O6.HCl) with respect to the labeled amount
＝ MS × AT/AS × V?/V × 1/C × 9
478 Benserazide Hydrochloride / Official Monographs JP XVII
259 C. precipitation does not dissolve.
drogen phosphate TS (pH 3.0), methanol and tetrahydrofu-
of Benserazide Hydrochloride in 10 mL of water, and per-
ran (65:27:8).
form the test with this solution as directed under Ultraviolet-
Flow rate: Adjust so that the retention time of benidipine
visible Spectrophotometry <2.24>: the absorbance of this so-
lution at 430 nm is not more than 0.10.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Benser-
azide Hydrochloride according to Method 2, and perform
ditions, the internal standard and benidipine are eluted in
less than 8.
exposure to light, using light-resistant vessels. Dissolve 0.25
g of Benserazide Hydrochloride in 10 mL of methanol, and
use this solution as the sample solution. Pipet 1 mL and 3
mL of the sample solution, add methanol to make exactly
the peak area of benidipine to that of the internal standard is
200 mL, and use these solutions as the standard solution (1)
not more than 1.0z.
and (2), respectively. Perform the test with these solutions as
Containers and storage Containers—Well-closed contain- directed under Thin-layer Chromatography <2.03>. Spot 2
ers. mL each of the sample solution and standard solutions (1)
and (2) on a plate of cellulose for thin-layer chromatogra-
phy. Develop the plate with a solution of formic acid in so-
Benserazide Hydrochloride dium chloride TS (1 in 1000) to a distance of about 10 cm,
and air-dry the plate. Spray evenly sodium carbonate TS,
ベンセラジド塩酸塩 air-dry, and then spray evenly Folin's TS on the plate: the
spots other than the principal spot from the sample solution
are not more intense than the spot from the standard solu-
tion (2), and the number of the spots which intense more
than the spot from the standard solution (1) are not more
than 2.
C10H15N3O5.HCl: 293.70 Water <2.48> Not more than 2.5z (0.5 g, volumetric titra-
(2RS )-2-Amino-3-hydroxy- tion, direct titration). Use a solution of salicylic acid in
N?-(2,3,4-trihydroxybenzyl)propanoylhydrazide methanol for water determination (3 in 20) instead of metha-
monohydrochloride nol for water determination.
[14919-77-8]
Benserazide Hydrochloride contains not less than Assay Weigh accurately about 0.3 g of Benserazide Hydro-
98.0z and not more than 101.0z of benserazide hy- chloride, dissolve in 5 mL of formic acid, add 50 mL of ace-
drochloride (C10H15N3O5.HCl), calculated on the an- tic acid (100), and titrate <2.50> immediately with 0.1 mol/L
hydrous basis. perchloric acid VS (potentiometric titration). Perform a
Description Benserazide Hydrochloride occurs as a white
to grayish white crystalline powder. Each mL of 0.1 mol/L perchloric acid VS
It is freely soluble in water and in formic acid, soluble in ＝ 29.37 mg of C10H15N3O5.HCl
methanol, very slightly soluble in ethanol (95).
It dissolves in 0.1 mol/L hydrochloric acid TS.
The pH of a solution of 1.0 g of Benserazide Hydrochlo-
It is hygroscopic.
It is gradually colored by light. Bentonite
A solution of Benserazide Hydrochloride (1 in 100) shows
ベントナイト
Bentonite is a natural, colloidal, hydrated aluminum
solution of Benserazide Hydrochloride in 0.1 mol/L hydro-
silicate.
chloric acid TS (1 in 10,000) as directed under Ultraviolet-
visible Spectrophotometry <2.24>, and compare the spectrum Description Bentonite occurs as a very fine, white to light
with the Reference Spectrum: both spectra exhibit similar in- yellow-brown powder. It is odorless. It has a slightly earthy
tensities of absorption at the same wavelengths. taste.
(2) Determine the infrared absorption spectrum of Ben- It is practically insoluble in water, in ethanol (95) and in
serazide Hydrochloride as directed in the potassium chloride diethyl ether.
disk method under Infrared Spectrophotometry <2.25>, and It swells in water.
Identification (1) Add 0.5 g of Bentonite to 3 mL of
diluted sulfuric acid (1 in 3), and heat until white fumes are
wave numbers.
evolved. Cool, add 20 mL of water, and filter. To 5 mL of
(3) To 10 mL of a solution of Benserazide Hydrochloride
the filtrate add 3 mL of ammonia TS: a white, gelatinous
(1 in 30) add silver nitrate TS: a white precipitate is formed.
precipitate is produced, which turns red on the addition of 5
To a portion of this precipitate add dilute nitric acid: the
JP XVII Official Monographs / Benzalkonium Chloride 479
drops of alizarin red S TS. 354.01), calculated on the anhydrous basis.

(2) Wash the residue obtained in (1) with water, add 2
Description Benzalkonium Chloride occurs as a white to
mL of a solution of methylene blue trihydrate (1 in 10,000),
yellowish white powder, colorless to light yellow, gelatinous
and wash again with water: the residue is blue in color.
pieces, or jelly-like fluid or mass. It has a characteristic
pH <2.54> To 1.0 g of Bentonite add 50 mL of water, and odor.
shake: the pH of the suspension is between 9.0 and 10.5. It is very soluble in water and in ethanol (95), and practi-
cally insoluble in diethyl ether.
Purity (1) Heavy metals <1.07>—To 1.5 g of Bentonite
A solution of Benzalkonium Chloride foams strongly
add 80 mL of water and 5 mL of hydrochloric acid, and boil
when shaken.
gently for 20 minutes with thorough stirring. Cool, centri-
fuge, collect the supernatant liquid, wash the residue with Identification (1) Dissolve 0.2 g of Benzalkonium Chlo-
two 10-mL portions of water, and centrifuge each. Combine ride in 1 mL of sulfuric acid, add 0.1 g of sodium nitrate,
the supernatant liquid and the washings, and add dropwise and heat for 5 minutes on a water bath. After cooling, add
ammonia solution (28). When a precipitate is produced, add 10 mL of water and 0.5 g of zinc powder, heat for 5 minutes,
dropwise dilute hydrochloric acid with vigorous stirring, and cool, and filter: the filtrate responds to the Qualitative Tests
dissolve. To the solution add 0.45 g of hydroxylammonium <1.09> for primary aromatic amines. The color of the solu-
chloride, and heat. Cool, and add 0.45 g of sodium acetate tion is red.
trihydrate, 6 mL of dilute acetic acid and water to make 150 (2) To 2 mL of a solution of Benzalkonium Chloride (1
mL. Pipet 50 mL of the solution, and perform the test using in 1000) add a mixture of 0.2 mL of a solution of bromophe-
this solution as the test solution. Prepare the control solution nol blue (1 in 2000) and 0.5 mL of sodium hydroxide TS: a
as follows: mix 2.5 mL of Standard Lead Solution, 0.15 g of blue color develops. Add 4 mL of chloroform to this solu-
hydroxylammonium chloride, 0.15 g of sodium acetate trihy- tion, and shake vigorously: the blue color shifts to the chlo-
drate, and 2 mL of dilute acetic acid, and add water to make roform layer. Collect the chloroform layer, and add drop-
50 mL (not more than 50 ppm). wise, with stirring, a solution of sodium lauryl sulfate (1 in
(2) Arsenic <1.11>—To 1.0 g of Bentonite add 5 mL of 1000): the chloroform layer turns colorless.
dilute hydrochloric acid, and gently heat to boil while stir- (3) Determine the absorption spectrum of a solution of
ring well. Cool immediately, and centrifuge. To the residue Benzalkonium Chloride in 0.1 mol/L hydrochloric acid TS
add 5 mL of dilute hydrochloric acid, shake well, and centri- (1 in 2000) as directed under Ultraviolet-visible Spectropho-
fuge. To the residue add 10 mL of water, and perform the tometry <2.24>, and compare the spectrum with the Refer-
same operations. Combine all the extracts, and heat on a ence Spectrum: both spectra exhibit similar intensities of ab-
water bath to concentrate to 5 mL. Perform the test with this sorption at the same wavelengths.
solution as the test solution (not more than 2 ppm). (4) To 1 mL of a solution of Benzalkonium Chloride (1
(3) Foreign matter—Place 2.0 g of Bentonite in a mor- in 100) add 2 mL of ethanol (95), 0.5 mL of dilute nitric acid
tar, add 20 mL of water to swell, disperse evenly with a pes- and 1 mL of silver nitrate TS: a white precipitate is pro-
tle, and dilute with water to 100 mL. Pour the suspension duced. This precipitate does not dissolve on the addition of
through a No. 200 (74 mm) sieve, and wash the sieve thor- dilute nitric acid, but dissolves on the addition of ammonia
oughly with water. No grit is felt when the fingers are rubbed TS.
over the wire mesh of the sieve.
Loss on drying <2.41> 5.0 – 10.0z (2 g, 1059C, 2 hours). of Benzalkonium Chloride in 10 mL of water: the solution is
clear and colorless to light yellow.
Gel formation Mix 6.0 g of Bentonite with 0.30 g of mag-
(2) Petroleum ether-soluble substances—To 3.0 g of
nesium oxide. Add the mixture, in several portions, to 200
Benzalkonium Chloride add water to make 50 mL, then add
mL of water contained in a glass-stoppered 500-mL cylinder.
50 mL of ethanol (99.5) and 5 mL of 0.5 mol/L sodium hy-
Agitate for 1 hour, transfer 100 mL of the suspension to a
droxide TS, and extract with three 50-mL portions of petro-
100-mL graduated cylinder, and allow to stand for 24 hours:
leum ether. Combine the petroleum ether extracts, and wash
not more than 2 mL of supernatant appears on the surface.
with three 50-mL portions of dilute ethanol. After shaking
Swelling power To 100 mL of water in a glass-stoppered well with 10 g of anhydrous sodium sulfate, filter through a
100-mL cylinder add 2.0 g of Bentonite in ten portions, dry filter paper, and wash the filter paper with two 10-mL
allowing each portion to settle before adding the next, and portions of petroleum ether. Evaporate the petroleum ether
allow to stand for 24 hours: the apparent volume of the sedi- on a water bath by heating, and dry the residue at 1059C for
ment at the bottom is not less than 20 mL. 1 hour: the residue is not more than 1.0z.
Containers and storage Containers—Well-closed contain- Water <2.48> Not more than 15.0z (volumetric titration,
ers. direct titration).
Benzalkonium Chloride Assay Weigh accurately about 0.15 g of Benzalkonium

Chloride, and dissolve in 75 mL of water. Adjust the pH be-
ベンザルコニウム塩化物 tween 2.6 and 3.4 by adding dropwise diluted dilute hydro-
chloric acid (1 in 2), add 1 drop of methyl orange TS, and
titrate <2.50> with 0.02 mol/L sodium tetraphenylboron VS
Benzalkonium Chloride is represented by the for
until the color of the solution becomes red.
mula [C6H5CH2N(CH3)2R]Cl, in which R extends
from C8H17 to C18H37, with C12H25 and C14H29 com- Each mL of 0.02 mol/L sodium tetraphenylboron VS
prising the major portion. ＝ 7.080 mg of C22H40ClN
It contains not less than 95.0z and not more than
105.0z of benzalkonium chloride (as C22H40ClN:
480 Benzalkonium Chloride Solution / Official Monographs JP XVII
and has a characteristic odor.
Benzalkonium Chloride Solution It is very soluble in water and in ethanol (95), and practi-
ベンザルコニウム塩化物液 A solution prepared by adding water to it vigorously
foams when shaken.
Benzalkonium Chloride Solution is an aqueous Identification (1) Dissolve 0.4 g of Benzalkonium Chlo-
solution containing not more than 50.0 w/vz of ben- ride Concentrated Solution 50 in 1 mL of sulfuric acid, add
zalkonium chloride. 0.1 g of sodium nitrate, and heat for 5 minutes on a water
It contains not less than 93.0z and not more than bath. After cooling, add 10 mL of water and 0.5 g of zinc
107.0z of the labeled amount of benzalkonium chlo- powder, heat for 5 minutes, cool, and filter: the filtrate re-
ride (C22H40ClN: 354.01). sponds to the Qualitative Tests <1.09> for primary aromatic
amines. The color of the solution is red.
Method of preparation Dissolve Benzalkonium Chloride in
(2) To 2 mL of a solution of Benzalkonium Chloride
Water, Purified Water or Purified Water in Containers. It is
Concentrated Solution 50 (1 in 500) add a mixture of 0.2 mL
also prepared by diluting Concentrated Benzalkonium Chlo-
of a solution of bromophenol blue (1 in 2000) and 0.5 mL of
ride Solution 50 with Water, Purified Water or Purified
sodium hydroxide TS: a blue color develops. Add 4 mL of
Water in Containers.
chloroform to this solution, and shake vigorously: the blue
Description Benzalkonium Chloride Solution is a clear, color shifts to the chloroform layer. Collect the chloroform
colorless to light yellow liquid, having a characteristic odor. layer, and add dropwise, with stirring, a solution of sodium
It foams strongly on shaking. lauryl sulfate (1 in 1000): the chloroform layer turns color-
less.
Identification (1) Evaporate a volume of Benzalkonium
Chloride Solution, equivalent to 0.2 g of Benzalkonium
Benzalkonium Chloride Concentrated Solution 50 in 0.1
Chloride, on a water bath to dryness, and proceed with the
mol/L hydrochloric acid TS (1 in 1000) as directed under
residue as directed in the Identification (1) under Benzalko-
nium Chloride.
the spectrum with the Reference Spectrum of Benzalkonium
(2) To a volume of Benzalkonium Chloride Solution,
Chloride: both spectra exhibit similar intensities of absorp-
equivalent to 0.01 g of Benzalkonium Chloride, add water to
tion at the same wavelengths.
make 10 mL. Proceed with 2 mL of this solution as directed
(4) To 1 mL of a solution of Benzalkonium Chloride
in the Identification (2) under Benzalkonium Chloride.
Concentrated Solution 50 (1 in 50) add 2 mL of ethanol (95),
(3) To a volume of Benzalkonium Chloride Solution,
0.5 mL of dilute nitric acid and 1 mL of silver nitrate TS: a
equivalent to 1 g of Benzalkonium Chloride, add water or
white precipitate is produced. This precipitate does not dis-
concentrate on a water bath, if necessary, to make 10 mL.
solve on the addition of dilute nitric acid, but dissolves on
To 1 mL of this solution add 0.1 mol/L hydrochloric acid
the addition of ammonia TS.
VS to make 200 mL, and proceed as directed in the Identifi-
cation (3) under Benzalkonium Chloride. Purity (1) Clarity and color of solution—Dissolve 2.0 g
(4) To a volume of Benzalkonium Chloride Solution, of Benzalkonium Chloride Concentrated Solution 50 in 10
equivalent to 0.1 g of Benzalkonium Chloride, add water or mL of water: the solution is clear and colorless to light yel-
concentrate on a water bath, if necessary, to make 10 mL. low.
Proceed with 1 mL of this solution as directed in the Identifi- (2) Petroleum ether-soluble substances—To 6.0 g of
cation (4) under Benzalkonium Chloride. Benzalkonium Chloride Concentrated Solution 50 add water
to make 50 mL, then add 50 mL of ethanol (99.5) and 5 mL
Assay Pipet a volume of Benzalkonium Chloride Solution,
of 0.5 mol/L sodium hydroxide TS, and extract with three
equivalent to about 0.15 g of benzalkonium chloride
50-mL portions of petroleum ether. Combine the petroleum
(C22H40ClN), dilute with water to make 75 mL, if necessary,
ether extracts, and wash with three 50-mL portions of dilute
and proceed as directed in the Assay under Benzalkonium
ethanol. After shaking well with 10 g of anhydrous sodium
Chloride.
sulfate, filter through a dry filter paper, and wash the filter
Each mL of 0.02 mol/L sodium tetraphenylboron VS paper with two 10-mL portions of petroleum ether.
＝ 7.080 mg of C22H40ClN Evaporate the petroleum ether on a water bath by heating,
and dry the residue at 1059C for 1 hour: the residue is not
more than 1.0z.
Benzalkonium Chloride Assay Weigh accurately about 0.3 g of Benzalkonium
Concentrated Solution 50 Chloride Concentrated Solution 50, and dissolve in 75 mL of
water. Adjust the pH to between 2.6 and 3.4 by adding
濃ベンザルコニウム塩化物液 50 dropwise diluted dilute hydrochloric acid (1 in 2), add 1 drop
of methyl orange TS, and titrate <2.50> with 0.02 mol/L
sodium tetraphenylboron VS until the color of the solution
Benzalkonium Chloride Concentrated Solution
becomes red.
50 is an aqueous solution, presented as
[C6H5CH2N(CH3)2R]Cl, where R ranges from C8H17 Each mL of 0.02 mol/L sodium tetraphenylborate VS
to C18H37, and mainly consisting of C12H25 and C14H29. ＝ 7.080 mg of C22H40ClN
It contains more than 50.0z and not more than
55.0z of benzalkonium chloride (C22H40ClN: 354.01).
Description Benzalkonium Chloride Concentrated Solu-
tion 50 is a colorless to light yellow liquid or jelly-like fluid,
JP XVII Official Monographs / Benzethonium Chloride 481

Benzbromarone directed under Thin-layer Chromatography <2.03>. Spot 10
ベンズブロマロン plate of silica gel with fluorescent indicator for thin-layer
chromatography. Develop the plate with a mixture of cyclo-
hexane, 4-methyl-2-pentanone, ethanol (99.5) and acetic acid
(100) (100:20:2:1) to a distance of about 15 cm, and air-dry
the plate. Examine under ultraviolet light (main wavelength:
254 nm): the spots other than the principal spot from the
sample solution are not more intense than the spot from the
standard solution.
Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum
C17H12Br2O3: 424.08
at a pressure not exceeding 0.67 kPa, phosphorus (V) oxide,
3,5-Dibromo-4-hydroxyphenyl 2-ethylbenzo[b]furan-3-yl
509C, 4 hours).
ketone
[3562-84-3] Residue on ignition <2.44> Not more than 0.1z (1 g).
Assay Weigh accurately about 0.6 g of Benzbromarone,
Benzbromarone, when dried, contains not less than
previously dried, dissolve in 30 mL of N, N-dimethylfor-
98.5z and not more than 101.0z of benzbromarone
mamide, and titrate <2.50> with 0.1 mol/L tetramethylam-
(C17H12Br2O3).
monium hydroxide VS (indicator: 5 drops of thymol blue-
Description Benzbromarone occurs as a white to light yel- dimethylformamide TS). Perform a blank determination,
low crystalline powder. and make any necessary correction.
It is very soluble in N, N-dimethylformamide, freely solu-
Each mL of 0.1 mol/L tetramethylammonium
ble in acetone, sparingly soluble in ethanol (99.5), and prac-
hydroxide VS
tically insoluble in water.
＝ 42.41 mg of C17H12Br2O3
It dissolves in dilute sodium hydroxide TS.
solution of Benzbromarone in 0.01 mol/L sodium hydroxide
TS (1 in 100,000) as directed under Ultraviolet-visible Spec-
trophotometry <2.24>, and compare the spectrum with the
Reference Spectrum: both spectra exhibit similar intensities Benzethonium Chloride
of absorption at the same wavelengths.
ベンゼトニウム塩化物
(2) Determine the infrared absorption spectrum of Benz-
bromarone, previously dried, as directed in the potassium
C27H42ClNO2: 448.08
Purity (1) Sulfate <1.14>—Dissolve 1.0 g of Benzbro- N-Benzyl-N, N-dimethyl-2-{2-[4-(1,1,3,3-
marone in 40 mL of acetone, and add 1 mL of dilute hydro- tetramethylbutyl)phenoxy]ethoxy}ethylaminium
chloric acid and water to make 50 mL. Perform the test chloride
using this solution as the test solution. Prepare the control [121-54-0]
solution as follows: to 0.40 mL of 0.005 mol/L sulfuric acid
VS add 40 mL of acetone, 1 mL of dilute hydrochloric acid Benzethonium Chloride, when dried, contains
and water to make 50 mL (not more than 0.019z). not less than 97.0z of benzethonium chloride
(2) Soluble halides—Dissolve 0.5 g of Benzbromarone in (C27H42ClNO2).
40 mL of acetone, and add 6 mL of dilute nitric acid and
Description Benzethonium Chloride occurs as colorless or
water to make 50 mL. Proceed with this solution as directed
white crystals. It is odorless.
under Chloride Limit Test <1.03>. Prepare the control solu-
It is very soluble in ethanol (95), freely soluble in water,
tion as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid
VS add 40 mL of acetone, 6 mL of dilute nitric acid and
A solution of Benzethonium Chloride foams strongly
water to make 50 mL.
when shaken.
(3) Heavy metals <1.07>—Proceed with 2.0 g of Benzbro-
marone according to Method 2, and perform the test. Pre- Identification (1) Dissolve 0.2 g of Benzethonium Chlo-
pare the control solution with 2.0 mL of Standard Lead So- ride in 1 mL of sulfuric acid, add 0.1 g of sodium nitrate,
lution (not more than 10 ppm). and heat for 5 minutes on a water bath. After cooling, add
(4) Iron <1.10>—Prepare the test solution with 1.0 g of 10 mL of water and 0.5 g of zinc powder, heat for 5 minutes,
Benzbromarone according to Method 3, and perform the test cool, and filter: the filtrate responds to the Qualitative Tests
according to Method A. Prepare the control solution with <1.09> for primary aromatic amines, developing a red color.
2.0 mL of Standard Iron Solution (not more than 20 ppm). (2) To 2 mL of a solution of Benzethonium Chloride
(5) Related substances—Dissolve 0.10 g of Benzbro- (1 in 1000) add a mixture of 0.2 mL of a solution of bromo-
marone in 10 mL of acetone, and use this solution as the phenol blue (1 in 2000) and 0.5 mL of sodium hydroxide TS:
sample solution. Pipet 1 mL of the sample solution, add ace- a blue color develops. Add 4 mL of chloroform to this solu-
tone to make exactly 100 mL, and use this solution as the tion, and shake vigorously: the blue color shifts to the chlo-
482 Benzethonium Chloride Solution / Official Monographs JP XVII
roform layer. Collect the chloroform layer, and add drop- under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-
wise a solution of sodium lauryl sulfate (1 in 1000) with its maxima between 262 nm and 264 nm, between 268 nm
stirring: the chloroform layer turns colorless. and 270 nm, and between 274 nm and 276 nm.
(3) Determine the absorption spectrum of a solution of (4) To a volume of Benzethonium Chloride Solution,
Benzethonium Chloride in 0.1 mol/L hydrochloric acid TS equivalent to 0.1 g of Benzethonium Chloride, add water, or
(1 in 5000) as directed under Ultraviolet-visible Spectropho- concentrate on a water bath, if necessary, to make 10 mL,
tometry <2.24>, and compare the spectrum with the Refer- and proceed with 1 mL of this solution as directed in the
ence Spectrum: both spectra exhibit similar intensities of ab- Identification (4) under Benzethonium Chloride.
Purity (1) Nitrite—Add 1.0 mL of Benzethonium Chlo-
(4) To 1 mL of a solution of Benzethonium Chloride (1
ride Solution to a mixture of 1 mL of a solution of glycine
in 100) add 2 mL of ethanol (95), 0.5 mL of dilute nitric acid
(1 in 10) and 0.5 mL of acetic acid (31): no gas is evolved.
and 1 mL of silver nitrate TS: a white precipitate is pro-
(2) Oxidizing substances—To 5 mL of Benzethonium
duced. This precipitate does not dissolve on addition of
Chloride Solution add 0.5 mL of potassium iodide TS and 2
dilute nitric acid, but dissolves on addition of ammonia TS.
to 3 drops of dilute hydrochloric acid: no yellow color is
Melting point <2.60> 158 – 1649C (after drying). produced.
Purity Ammonium—Dissolve 0.10 g of Benzethonium Assay Pipet a volume of Benzethonium Chloride Solution,
Chloride in 5 mL of water, and boil with 3 mL of sodium hy- equivalent to about 0.2 g of benzethonium chloride
droxide TS: the evolving gas dose not change moistened red (C27H42ClNO2), dilute with water to make 75 mL, if neces-
litmus paper to blue. sary, and proceed as directed in the Assay under Benzetho-
nium Chloride.
4 hours). Each mL of 0.02 mol/L sodium tetraphenylboron VS
Assay Weigh accurately about 0.2 g of Benzethonium
Chloride, previously dried, dissolve in 75 mL of water, add
diluted dilute hydrochloric acid (1 in 2) dropwise to adjust
the pH to 2.6–3.4, then add 1 drop of methyl orange TS, and
titrate <2.50> with 0.02 mol/L tetraphenylboron VS until the Benzoic Acid
solution develops a red.
安息香酸
Each mL of 0.02 mol/L sodium tetraphenylboron VS
C7H6O2: 122.12
Benzoic acid
[65-85-0]
Benzethonium Chloride Solution
ベンゼトニウム塩化物液
Benzoic Acid, when dried, contains not less than
99.5z of benzoic acid (C7H6O2).
Description Benzoic Acid occurs as white crystals or crys-
Benzethonium Chloride Solution contains not less
talline powder. It is odorless, or has a faint, benzaldehyde-
like odor.
amount of benzethonium chloride (C27H42ClNO2:
It is freely soluble in ethanol (95), in acetone and in diethyl
448.08).
ether, soluble in hot water, and slightly soluble in water.
Method of preparation Dissolve Benzethonium Chloride in
Identification Dissolve 1 g of Benzoic Acid in 8 mL of so-
Water, Purified Water or Purified Water in Containers.
dium hydroxide TS, and add water to make 100 mL. This
Description Benzethonium Chloride Solution is a clear, solution responds to the Qualitative Tests <1.09> (2) for ben-
colorless liquid. It is odorless. zoate.
It foams strongly when shaken.
Melting point <2.60> 121 – 1249
C
Identification (1) Evaporate a volume of Benzethonium
Purity (1) Heavy metals <1.07>—Dissolve 1.0 g of Ben-
Chloride Solution, equivalent to 0.2 g of Benzethonium
zoic Acid in 25 mL of acetone, add 2 mL of dilute acetic acid
Chloride, on a water bath to dryness, and proceed with the
and water to make 50 mL, and perform the test using this
residue as directed in the Identification (1) under Benzetho-
solution as the test solution. Prepare the control solution as
nium Chloride.
follows: to 2.0 mL of Standard Lead Solution add 2 mL of
(2) To a volume of Benzethonium Chloride Solution,
dilute acetic acid, 25 mL of acetone and water to make 50
equivalent to 0.01 g of Benzethonium Chloride, add water to
mL (not more than 20 ppm).
make 10 mL, proceed with 2 mL of this solution as directed
(2) Chlorinated compounds—Take 0.5 g of Benzoic Acid
in the Identification (2) under Benzethonium Chloride.
and 0.7 g of calcium carbonate in a crucible, mix with a
(3) To a volume of Benzethonium Chloride Solution,
small amount of water, and dry. Ignite it at about 6009C,
equivalent to 1 g of Benzethonium Chloride, and add water
dissolve in 20 mL of dilute nitric acid, and filter. Wash the
or concentrate on a water bath to make 10 mL. To 1 mL of
residue with 15 mL of water, combine the filtrate and the
this solution add 0.1 mol/L hydrochloric acid TS to make
washing, add water to make 50 mL, and add 0.5 mL of silver
500 mL, and determine the abosprition spectrum as directed
nitrate TS: this solution has not more turbid than the follow-
JP XVII Official Monographs / Benzyl Alcohol 483
ing control solution. It is miscible with ethanol (95), with fatty oils and with
Control solution: Dissolve 0.7 g of calcium carbonate in essential oils.
20 mL of dilute nitric acid, and filter. Wash the residue with It is soluble in water.
15 mL of water, combine the filtrate and the washings, add Specific gravity d 20
20: 1.043 – 1.049
1.2 mL of 0.01 mol/L hydrochloric acid VS and water to 
Identification Determine the infrared absorption spec-
make 50 mL, and add 0.5 mL of silver nitrate TS.
trum of Benzyl Alcohol as directed in the liquid film method
(3) Potassium permanganate-reducing substances—Add
0.02 mol/L potassium permanganate VS dropwise to a boil-
ing mixture of 100 mL of water and 1.5 mL of sulfuric acid,
similar intensities of absorption at the same wave numbers.
until a red color persists for 30 seconds. Dissolve 1.0 g of
Benzoic Acid in this boiling solution, and add 0.50 mL of Refractive index <2.45> n 20
D : 1.538 – 1.541
0.02 mol/L potassium permanganate VS: a red color persists
Purity (1) Clarity and color of solution—Dissolve 2.0
for at least 15 seconds.
mL of Benzyl Alcohol in 60 mL of water: the solution is
(4) Phthalic acid—To 0.10 g of Benzoic Acid add 1 mL
clear and colorless.
of water and 1 mL of resorcinol-sulfuric acid TS, and heat
(2) Acidity—To 10 mL of Benzyl Alcohol add 10 mL of
the mixture in an oil bath heated at a temperature between
ethanol (95) and 2 drops of phenolphthalein TS, and add
1209C and 1259 C. After evaporating the water, heat the
dropwise 0.1 mol/L sodium hydroxide VS until the solution
residue for 90 minutes, cool, and dissolve in 5 mL of water.
acquires a light red color: the amount of 0.1 mol/L sodium
To 1 mL of the solution add 10 mL of a solution of sodium
hydroxide VS used is not more than 1.0 mL.
hydroxide (43 in 500), shake, then examine under light at a
(3) Benzaldehyde and other related substances—Use
wavelength between 470 nm and 490 nm: the green fluores-
Benzyl Alcohol as the sample solution. Separately, dissolve
cence of the solution is not more intense than that of the fol-
exactly 0.100 g of ethylbenzene in Benzyl Alcohol to make
lowing control solution.
exactly 10 mL. Pipet 2 mL of this solution, add Benzyl Alco-
Control solution: Dissolve 61 mg of potassium hydrogen
hol to make exactly 20 mL, and use this solution as the ethyl-
phthalate in water to make exactly 1000 mL. Measure ex-
benzene stock solution. Separately, dissolve exactly 2.000 g
actly 1 mL of the solution, add 1 mL of resorcinol-sulfuric
of dicyclohexyl in Benzyl Alcohol to make exactly 10 mL.
acid TS, and proceed as directed above.
Pipet 2 mL of this solution, add Benzyl Alcohol to make ex-
actly 20 mL, and use this solution as the dicyclohexyl stock
test with 0.5 g of Benzoic Acid. The solution is not more
solution. Separately, weigh exactly 0.750 g of benzaldehyde
colored than Matching Fluid Q.
and 0.500 g of cyclohexylmethanol, and add Benzyl Alcohol
Loss on drying <2.41> Not more than 0.5z (1 g, silica gel, to make exactly 25 mL. Pipet 1 mL of this solution, add ex-
3 hours). actly 2 mL of ethylbenzene stock solution and exactly 3 mL
of dicyclohexyl stock solution, then add Benzyl Alcohol to
Assay Weigh accurately about 0.5 g of Benzoic Acid, pre- solution (1). Perform the test with exactly 0.1 mL each of the
viously dried, dissolve in 25 mL of neutralized ethanol and sample solution and standard solution (1) as directed under
25 mL of water, and titrate <2.50> with 0.1 mol/L sodium Gas Chromatography <2.02> according to the following con-
hydroxide VS (indicator: 3 drops of phenolphthalein TS). ditions, and when the peak having the retention time cor-
responding to ethylbenzene and dicyclohexyl appears on the
chromatogram obtained with the sample solution, correct
＝ 12.21 mg of C7H6O2
the peak areas of ethylbenzene and dicyclohexyl obtained
Containers and storage Containers—Well-closed contain- with the standard solution (1) by deducting the relevant peak
ers. area obtained with the sample solution, the peak area of ben-
zaldehyde obtained with the sample solution is not more
than the difference between the peak areas of benzaldehyde
Benzyl Alcohol of the sample solution and the standard solution (1)
(0.15z), and the peak area of cyclohexylmethanol with the
ベンジルアルコール sample solution is not more than the difference between the
peak areas of cyclohexylmethanol of the sample solution and
the standard solution (1) (0.10z). The total area of the
peaks having smaller retention time than benzyl alcohol and
other than benzaldehyde and cyclohexylmethanol obtained
C7H8O: 108.14 with the sample solution is not more than 4 times the peak
Benzyl alcohol area or the corrected peak area of ethylbenzene with the
[100-51-6] standard solution (1) (0.04z). The total area of the peaks
having larger retention time than benzyl alcohol obtained
This monograph is harmonized with the European Phar-
with the sample solution is not more than the peak area or
macopoeia and the U.S. Pharmacopeia. The parts of the text
the corrected peak area of dicyclohexyl with the standard so-
that are not harmonized are marked with symbols ( ).
lution (1) (0.3z). For these calculations the peak areas less
than 1/100 times the peak area or the corrected peak area of
Benzyl Alcohol contains not less than 98.0z and
ethylbenzene with the standard solution (1) are excluded.
not more than 100.5z of benzyl alcohol (C7H8O).
 Benzyl Alcohol labeled that it is suitable for use in the
The label states, where applicable, that it is suita-
manufacture of injection forms meets the following require-
ble for use in the manufacture of injection forms.
ments.
Description Benzyl Alcohol is a clear, colorless oily liq- Use Benzyl Alcohol as the sample solution. Separately,
uid. weigh exactly 0.250 g of benzaldehyde and 0.500 g of cyclo-
484 Benzyl Benzoate / Official Monographs JP XVII
hexylmethanol, and add Benzyl Alcohol to make exactly 25 conical flask. Add 0.5 mL of saturated potassium iodide so-
mL. Pipet 1 mL of this solution, add exactly 2 mL of the lution, shake for exactly 1 minute, add 30 mL of water, and
ethylbenzene stock solution and exactly 2 mL of the dicyclo- titrate <2.50> with 0.01 mol/L sodium thiosulfate VS, adding
hexyl stock solution, then add Benzyl Alcohol to make ex- the titrant slowly with continuous vigorous shaking, until the
actly 20 mL, and use this solution as the standard solution blue color of the solution disappears after addition of 5 mL
(2). Perform the test with exactly 0.1 mL each of the sample of starch TS near the end point where the solution is a pale
solution and standard solution (2) as directed under Gas yellow color. Perform a blank determination in the same
Chromatography <2.02> according to the following condi- manner. Calculate the amount of peroxide by the following
tions, and when the peak having the retention time cor- formula: not more than 5. In the determination, the required
responding to ethylbenzene and dicyclohexyl appears on the amount of 0.01 mol/L sodium thiosulfate VS must not
chromatogram obtained with the sample solution, correct exceed 0.1 mL.
the peak areas of ethylbenzene and dicyclohexyl obtained
Amount (mEq/kg) of peroxide ＝ 10 × (V1 － V0)/M
with the standard solution (2) by deducting the relevant peak
area obtained with the sample solution, the peak area of V1: Volume (mL) of 0.01 mol/L sodium thiosulfate VS
benzaldehyde obtained with the sample solution is not more consumed in the test
than the difference between the peak areas of benzaldehyde V0: Volume (mL) of 0.01 mol/L sodium thiosulfate VS
of the sample solution and the standard solution (2) consumed in the blank determination
(0.05z), and the peak area of cyclohexylmethanol with the M: Amount (g) of Benzyl Alcohol taken
sample solution is not more than the difference between the
(5) Residue on evaporation—Perform the test after
peak areas of cyclohexylmethanol of the sample solution and
conformation that the sample meets the requirement of the
the standard solution (2) (0.10z). The total area of the
peroxide value. Transfer 10.0 g of Benzyl Alcohol to a
peaks having smaller retention time than benzyl alcohol and
porcelain or quartz crucible or platinum dish, previously
other than benzaldehyde and cyclohexylmethanol obtained
weighed accurately, and heat on a hot-plate at not exceeding
with the sample solution is not more than 2 times the peak
2009C, taking care to avoid boiling, to evaporate to dryness.
area or the corrected peak area of ethylbenzene with the
Dry the residue on the hot-plate for 1 hour, and allow to
standard solution (2) (0.02z). The total area of the peaks
cool in a desiccator: not more than 5 mg.
having larger retention time than benzyl alcohol obtained
with the sample solution is not more than the peak area of or Assay Weigh accurately about 0.9 g of Benzyl Alcohol,
the corrected peak area dicyclohexyl with the standard solu- add exactly 15 mL of a freshly prepared mixture of de-
tion (2) (0.2z). For these calculation the peak areas less than hydrated pyridine and acetic anhydride (7:1), and heat on a
1/100 times the peak area or the corrected peak area of water bath under a reflux condenser for 30 minutes. Cool,
ethylbenzene with the standard solution (2) are excluded. add 25 mL of water, and titrate <2.50> the excess acetic acid
Operating conditions— with 1 mol/L sodium hydroxide VS (indicator: 2 drops of
Detector: A hydrogen flame-ionization detector. phenolphthalein TS). Perform a blank determination.
Column: A fused silica column 0.32 mm in inside diameter
and 30 m in length, coated inside with polyethylene glycol
＝ 108.1 mg of C7H8O
20 M for gas chromatography in 0.5 mm thickness.
Column temperature: Inject at a constant temperature of Containers and storage Containers—Tight containers.
about 509C, raise the temperature at a rate of 59C per Storage—Light-resistant.
minute to 2209C, and maintain at 2209C for 35 minutes.
Temperature of injection port: A constant temperature of
about 2009C. Benzyl Benzoate
Temperature of detector: A constant temperature of about
3109C. 安息香酸ベンジル
Flow rate: 25 cm/second.
Split ratio: Splitless.
Detection sensitivity: When 0.1 mL of the standard solu-
tion (1) is injected, adjust the sensitivity of the detector so
that the height of the peak of ethylbenzene is not less than C14H12O2: 212.24
30z of the full scale of the recorder. For Benzyl Alcohol Benzyl benzoate
labeled to use for injection, use the standard solution (2) [120-51-4]
instead of the standard solution (1).
System suitability— Benzyl Benzoate contains not less than 99.0z of
System performance: When the procedure is run with the benzyl benzoate (C14H12O2).
standard solution (1) under the above operating conditions,
Description Benzyl Benzoate is a colorless, clear, viscous
the retention time of benzyl alcohol is about 26 minutes, the
liquid. It has a faint, aromatic odor and a pungent, burning
relative retention times of ethylbenzene, dicyclohexyl, ben-
taste.
zaldehyde and cyclohexylmethanol to benzyl alcohol are
It is miscible with ethanol (95) and with diethyl ether.
about 0.28, about 0.59, about 0.68 and about 0.71, respec-
It is practically insoluble in water.
tively, and the resolution between the peaks of benzaldehyde
Congealing point: about 179C
and cyclohexylmethanol is not less than 3.0. In the case of
20: about 1.123
Benzyl Alcohol labeled to use for injection, proceed with the
Boiling point: about 3239 C
standard solution (2) instead of the standard solution (1).
(4) Peroxide value—Weigh accurately about 5 g of Ben- Identification (1) Heat gently 1 mL of Benzyl Benzoate
zyl Alcohol, and dissolve in 30 mL of a mixture of acetic with 5 mL of sodium carbonate TS and 2 mL of potassium
acid (100) and chloroform (3:2) in a 250-mL glass-stoppered permanganate TS: the odor of benzaldehyde is perceptible.
JP XVII Official Monographs / Benzylpenicillin Benzathine Hydrate 485
(2) Warm the titrated mixture obtained in the Assay on a (2) Determine the infrared absorption spectrum of Ben-
water bath to remove ethanol, and add 0.5 mL of iron (III) zylpenicillin Benzathine Hydrate as directed in the potassium
chloride TS: a light yellow-red precipitate is produced, which bromide disk method under Infrared Spectrophotometry
turns white on the addition of dilute hydrochloric acid. <2.25>, and compare the spectrum with the Reference Spec-
Refractive index <2.45> n 20
D : 1.568 – 1.570
Purity Acidity—Dissolve 5.0 mL of Benzyl Benzoate in 25
D : ＋217 – ＋2339(0.1 g calcu-
mL of neutralized ethanol, and add 0.50 mL of 0.1 mol/L
lated on the anhydrous basis, methanol, 20 mL, 100 mm).
sodium hydroxide VS: a red color develops.
Benzylpenicillin Benzathine Hydrate according to Method 2,
Assay Weigh accurately about 2 g of Benzyl Benzoate, add and perform the test. Prepare the control solution with 2.0
exactly 50 mL of 0.5 mol/L potassium hydroxide-ethanol mL of Standard Lead Solution (not more than 20 ppm).
VS, and boil gently for 1 hour under a reflux condenser with (2) Arsenic <1.11>—Prepare the test solution with 1.0 g
a carbon dioxide-absorbing tube (soda lime). Cool, and of Benzylpenicillin Benzathine Hydrate according to Method
titrate <2.50> the excess potassium hydroxide with 0.5 mol/L 3, and perform the test (not more than 2 ppm).
hydrochloric acid VS (indicator: 2 drops of phenolphthalein (3) Related substances—Dissolve 70 mg of Benzylpenicil-
TS). Perform a blank determination. lin Benzathine Hydrate in 25 mL of methanol, add a solution
prepared by dissolving 1.02 g of disodium hydrogen phos-
Each mL of 0.5 mol/L potassium hydroxide-ethanol VS
phate and 6.80 g of potassium dihydrogen phosphate in
＝ 106.1 mg of C14H12O2
water to make 1000 mL to make 50 mL, and use this solution
Containers and storage Containers—Tight containers. as the sample solution. Pipet 1 mL of the sample solution,
Storage—Light-resistant. add the mobile phase A to make exactly 100 mL, and use this
exactly 20 mL each of the sample solution and standard
Benzylpenicillin Benzathine Hydrate solution as directed under Liquid Chromatography <2.01>
ベンジルペニシリンベンザチン水和物 peak area by the automatic integration method: the area of
the peak having the relative retention time of about 2.4 to
benzylpenicillin obtained from the sample solution is not
larger than 2 times the total area of the peaks of benzyl-
penicillin and N, N?-dibenzylethylenediamine obtained from
the standard solution, and the area of the peak other than
benzylpenicillin, N, N?-dibenzylethylenediamine and the
peak having the relative retention time of about 2.4 to
(C16H18N2O4S)2.C16H20N2.4H2O: 981.18
benzylpenicillin is not larger than the total area of the peaks
(2S,5R,6R)-3,3-Dimethyl-7-oxo-6-[(phenylacetyl)amino]-4-thia-
of benzylpenicillin and N, N?-dibenzylethylenediamine from
1-azabicyclo[3.2.0]heptane-2-carboxylic acid
the standard solution.
hemi(N, N?-dibenzylethane-1,2-diamine)dihydrate
[41372-02-5]
length: 220 nm).
Benzylpenicillin Benzathine Hydrate is the N, N?-
dibenzylethylenediamine salt of a penicillin compound
having antibacterial activity produced by the growth
of Penicillium species.
It contains not less than 1213 Units and not more
409C.
than 1333 Units per mg, calculated on the anhydrous
Mobile phase A: A mixture of water, methanol and 0.25
basis. The potency of Benzylpenicillin Benzathine Hy-
mol/L potassium dihydrogen phosphate TS (pH 3.5) (6:3:1).
drate is expressed as unit calculated from the amount
Mobile phase B: A mixture of methanol, water and 0.25
of benzylpenicillin sodium (C16H17N2NaO4S: 356.37).
mol/L potassium dihydrogen phosphate TS (pH 3.5) (6:3:1).
1 Unit of Benzylpenicillin Benzathine Hydrate is
equivalent to 0.6 mg of benzylpenicillin sodium
(C16H17N2NaO4S). It contains not less than 24.0z and
not more than 27.0z of N, N?-dibenzylethylenedia-
mine (C16H20N2: 240.34), calculated on the anhydrous Time after injection Mobile phase A Mobile phase B
basis. of sample (min) (volz) (volz)
Description Benzylpenicillin Benzathine Hydrate occurs as 0 – 10 75 25

a white crystalline powder. 10 – 20 75 → 0 25 → 100
It is slightly soluble in methanol and in ethanol (99.5), and 20 – 55 0 100
Identification (1) Determine the absorption spectrum of a Flow rate: 1.0 mL per minute.
solution of Benzylpenicillin Benzathine Hydrate in methanol Time span of measurement: About 3 times as long as the
(1 in 2000) as directed under Ultraviolet-visible Spectropho- retention time of benzylpenicillin, beginning after the solvent
tometry <2.24>, and compare the spectrum with the Refer- peak.
ence Spectrum: both spectra exhibit similar intensities of ab- System suitability—
sorption at the same wavelengths. Test for required detectability: To exactly 1 mL of the
486 Benzylpenicillin Potassium / Official Monographs JP XVII
standard solution add the mobile phase A to make exactly 20 with 20 mL of the standard solution under the above operat-
mL. Confirm that the peak area of benzylpenicillin obtained ing conditions, the relative standard deviations of the peak
from 20 mL of this solution is equivalent to 3.5 to 6.5z of areas of N, N?-dibenzylethylenediamine and benzylpenicillin
that obtained from the standard solution. are not more than 2.0z, respectively.
System performance: When the procedure is run with 20 (2) N, N?-Dibenzylethylenediamine—Determine the
mL of the standard solution under the above operating con- areas, AT and AS, of the peak corresponding to N, N?-
ditions, N, N?-dibenzylethylenediamine and benzylpenicillin dibenzylethylenediamine on the chromatograms obtained in
are eluted in this order with the resolution between these (1) with the sample solution and standard solution.
Amount (z) of N, N?-dibenzylethylenediamine (C16H20N2)
＝ MS/MT × AT/AS × 100 × 0.667
ing conditions, the relative standard deviation of the peak MS: Amount (mg) of N, N?-dibenzylethylenediamine dia-
areas of benzylpenicillin is not more than 2.0z. cetate taken
MT: Amount (mg) of Benzylpenicillin Benzathine Hydrate
Water <2.48> 5.0 – 8.0z (1 g, volumetric titration, direct
taken
titration).
0.667: Conversion factor for the molecular mass of N, N?-
Assay (1) Benzylpenicillin—Weigh accurately an amount dibenzylethylenediamine diacetate (C16H20N2･
of Benzylpenicillin Benzathine Hydrate, equivalent to about 2CH3COOH) to that of N, N?-dibenzylethylenedia-
85,000 Units, dissolve in 25 mL of methanol, and add a solu- mine (benzathine, C16H20N2)
tion containing 1.02 g of disodium hydrogen phosphate and
6.80 g of potassium dihydrogen phosphate in 1000 mL of
add a mixture of the solution containing 1.02 g of disodium
hydrogen phosphate and 6.80 g of potassium dihydrogen
phosphate in 1000 mL of water and methanol (1:1) to make Benzylpenicillin Potassium
Separately, weigh accurately an amount of Benzylpenicillin
Penicillin G Potassium
Potassium RS, equivalent to about 85,000 Units, and about
ベンジルペニシリンカリウム
25 mg of N, N?-dibenzylethylenediamine diacetate, dissolve
in 25 mL of methanol, and add the solution containing
1.02 g of disodium hydrogen phosphate and 6.80 g of potas-
sium dihydrogen phosphate in 1000 mL of water to make
exactly 50 mL. Pipet 5 mL of this solution, add a mixture of
the solution containing 1.02 g of disodium hydrogen phos-
phate and 6.80 g of potassium dihydrogen phosphate in 1000
mL of water and methanol (1:1) to make exactly 20 mL, and C16H17KN2O4S: 372.48
use this solution as the standard solution. Perform the test Monopotassium (2S,5R,6R)-3,3-dimethyl-
with exactly 20 mL each of the sample solution and standard 7-oxo-6-[(phenylacetyl)amino]-4-thia-
solution as directed under Liquid Chromatography <2.01> 1-azabicyclo[3.2.0]heptane-2-carboxylate
according to the following conditions, and determine the [113-98-4]
peak areas, AT ands AS, of benzylpenicillin in each solution.
Benzylpenicillin Potassium is the potassium salt of a
Amount (unit) of benzylpenicillin sodium (C16H17N2NaO4S)
＝ MS × AT/AS
penicillin substance having antibacterial activity pro-
duced by the growth of Penicillium species.
MS: Amount (unit) of Benzylpenicillin Potassium RS It contains not less than 1430 units and not more
taken than 1630 units per mg, calculated on the dried
basis. The potency of Benzylpenicillin Potassium is
expressed as mass unit of benzylpenicillin potassium
(C16H17KN2O4S). One unit of Benzylpenicillin Potas-
length: 220 nm).
sium is equivalent to 0.63 mg of benzylpenicillin potas-
sium.
gel for liquid chromatography (5 mm in particle diameter). Description Benzylpenicillin Potassium occurs as white,
Column temperature: A constant temperature of about crystals or crystalline powder.
409 C. It is very soluble in water, and slightly soluble in ethanol
Mobile phase: A mixture of water, methanol and 0.25 (99.5).
mol/L potassium dihydrogen phosphate TS (pH 3.5)
(11:7:2).
solution of Benzylpenicillin Potassium (1 in 1000) as directed
Flow rate: Adjust so that the retention time of benzyl-
penicillin is about 18 minutes.
spectrum of a solution of Benzylpenicillin Potassium RS
prepared in the same manner as the sample solution: both
ditions, N, N?-dibenzylethylenediamine and benzylpenicillin
wavelengths.
are eluted in this order with the resolution between these
(2) Determine the infrared absorption spectrum of Ben-
zylpenicillin Potassium as directed in the potassium bromide
JP XVII Official Monographs / Benzylpenicillin Potassium for Injection 487
compare the spectrum with the Reference Spectrum or the about 6 × 104 Units, dissolve each in water to make exactly
spectrum of Benzylpenicillin Potassium RS: both spectra 20 mL, and use these solutions as the sample solution and
exhibit similar intensities of absorption at the same wave the standard solution, respectively. Perform the test with
numbers. exactly 20 mL each of the sample solution and standard
(3) Benzylpenicillin Potassium responds to the Qualita- solution as directed under Liquid Chromatography <2.01>
tive Tests <1.09> (1) for potassium salt. according to the following conditions, and determine the
peak areas, AT and AS, of benzylpenicillin in each solution.
Optical rotation <2.49> [a]20D : ＋270 – ＋3009(1.0 g calcu-
lated on the dried basis, water, 50 mL, 100 mm). Amount (unit) of benzylpenicillin potassium
(C16H17KN2O4S)
＝ M S × A T / AS
1.0 g of Benzylpenicillin Potassium in 100 mL of water is be-
tween 5.0 and 7.5. MS: Amount (unit) of benzylpenicillin potassium RS taken
Purity (1) Clarity and color of solution—A solution ob- Operating conditions—
tained by dissolving 1.0 g of Benzylpenicillin Potassium in Detector: An ultraviolet absorption photometer (wave-
10 mL of water is clear, and its absorbance at 400 nm deter- length: 254 nm).
mined as directed under Ultraviolet-visible Spectrophotome- Column: A stainless steel column 4.6 mm in inside diame-
try <2.24> is not more than 0.10. ter and 25 cm in length, packed with octadecylsilanized silica
(2) Heavy metals <1.07>—Proceed with 2.0 g of Benzyl- gel for liquid chromatography (7 mm in particle diameter).
penicillin Potassium according to Method 4, and perform Column temperature: A constant temperature of about
the test. Prepare the control solution with 2.0 mL of Stand- 259C.
ard Lead Solution (not more than 10 ppm). Mobile phase: A mixture of diammonium hydrogen phos-
(3) Arsenic <1.11>—Prepare the test solution by inciner- phate solution (33 in 5000) and acetonitrile (19:6), adjusted
ating 1.0 g of Benzylpenicillin Potassium according to to pH 8.0 with phosphoric acid.
Method 4, and perform the test. In the incineration, use a Flow rate: Adjust so that the retention time of benzyl-
crucible of porcelain, and after addition of 10 mL of a solu- penicillin is about 7.5 minutes.
tion of magnesium nitrate hexahydrate in ethanol (95) (1 in System suitability—
10) add 1 mL of hydrogen peroxide (30), then burn the System performance: Dissolve 40 mg of Benzylpenicillin
ethanol (not more than 2 ppm). Potassium in 20 mL of water. Separately, dissolve 10 mg of
(4) Related substances—Dissolve 40 mg of Benzylpenicil- methyl parahydroxybenzoate in 20 mL of acetonitrile. To
lin Potassium in 20 mL of water, and use this solution as the 1 mL of this solution add water to make 20 mL. Mix 1 mL
sample solution. Pipet 1 mL of the sample solution, add each of these solutions, and add water to make 100 mL.
water to make exactly 100 mL, and use this solution as the When the procedure is run with 20 mL of this solution under
standard solution. Perform the test with exactly 20 mL each the above operating conditions, benzylpenicillin and methyl
of the sample solution and standard solution as directed parahydroxybenzoate are eluted in this order with the resolu-
under Liquid Chromatography <2.01> according to the fol- tion between these peaks being not less than 8.
lowing conditions, and determine each peak area by the System repeatability: When the test is repeated 6 times
automatic integration method: the area of the peaks other with 20 mL of the standard solution under the above operat-
than benzylpenicillin obtained from the sample solution is ing conditions, the relative standard deviation of the peak
not larger than the peak area of benzylpenicillin obtained area of benzylpenicillin is not more than 1.0z.
from the standard solution, and the total area of the peaks
other than benzylpenicillin from the sample solution is not
larger than 3 times the peak area of benzylpenicillin from the
standard solution.
Operating conditions— Benzylpenicillin Potassium for
Detector, column, column temperature, mobile phase, and Injection
the Assay. 注射用ベンジルペニシリンカリウム
retention time of benzylpenicillin.
Benzylpenicillin Potassium for Injection is a prepa-
ration for injection which is dissolved before use.
System Performance: Proceed as directed in the system
107.0z of the labeled potency of benzylpenicillin po-
tassium (C16H17KN2O4S: 372.48).
ard solution, and add water to make exactly 100 mL. Con-
firm that the peak area of benzylpenicillin obtained from 20 Method of preparation Prepare as directed under Injec-
mL of this solution is equivalent to 7 to 13z of that obtained tions, with Benzylpenicillin Potassium.
Description Benzylpenicillin Potassium for Injection oc-
curs as white, crystals or crystalline powder.
ing conditions, the relative standard deviation of the peak Identification Proceed as directed in the Identification (2)
area of benzylpenicillin is not more than 2.0z. under Benzylpenicillin Potassium.
Loss on drying <2.41> Not more than 1.0z (3 g, reduced Osmotic pressure ratio Being specified separately when the
pressure not exceeding 0.67 kPa, 609C, 3 hours). drug is granted approval based on the Law.
Assay Weigh accurately amounts of Benzylpenicillin pH <2.54> The pH of a solution prepared by dissolving an
Potassium and Benzylpenicillin Potassium RS, equivalent to amount of Benzylpenicillin Potassium for Injection, equiva-
488 Bepotastine Besilate / Official Monographs JP XVII
lent to 1.0 × 105 Units of Benzylpenicillin Potassium, in 10 conditions, the relative standard deviation of the peak area
mL of water is 5.0 to 7.5. of benzylpenicillin is not more than 1.0z.
Purity Clarity and color of solution—A solution prepared Containers and storage Containers—Hermetic containers.
by dissolving an amount of Benzylpenicillin Potassium for
Injection, equivalent to 1.0 × 106 Units of Benzylpenicillin
Potassium, in 10 mL of water is clear. Perform the test with Bepotastine Besilate
this solution as directed under Ultraviolet-visible Spectro-
photometry <2.24>: the absorbance at 400 nm is not more ベポタスチンベシル酸塩
than 0.10.
C, 3 hours).
um, below 0.67 kPa, 609
Bacterial endotoxins <4.01> Less than 1.25 × 10－4 EU/
Unit.
C21H25ClN2O3.C6H6O3S: 547.06
(S)-4-s
4-[(4-Chlorophenyl)(pyridin-2-yl)methoxy]piperidin-
Foreign insoluble matter <6.06> Perform the test according 1-yltbutanoic acid monobenzenesulfonate
to Method 2: it meets the requirement. [190786-44-8]
Bepotastine Besilate contains not less than 99.0z
ment.
and not more than 101.0z of bepotastine besilate
Sterility <4.06> Perform the test according to the Mem- (C21H25ClN2O3.C6H6O3S), calculated on the anhydrous
brane filtration method: it meets the requirement. and residual solvent-free basis.
Assay Weigh accurately the mass of the contents of not less Description Bepotastine Besilate occurs as white to pale
than 10 containers of Benzylpenicillin Potassium for Injec- yellowish white, crystals or crystalline powder.
tion. Weigh accurately an amount of a portion of the con- It is very soluble in acetic acid (100), and sparingly soluble
tents, equivalent to about 6 × 104 Units of Benzylpenicillin in water and in ethanol (99.5).
Potassium, dissolve in water to make exactly 20 mL, and use The pH of a solution of 1 g of Bepotastine Besilate in 100
this solution as the sample solution. Separately, weigh accu- mL of water is about 3.8.
rately an amount of Benzylpenicillin Potassium RS, equiva-
lent to about 6 × 104 Units, dissolve in water to make ex-
solution of Bepotastine Besilate (1 in 20,000) as directed
tography <2.01> according to the following conditions. De-
wavelengths.
termine the peak areas, AT and AS, of benzylpenicillin in
each solution.
Bepotastine Besilate as directed in the potassium bromide
Amount (unit) of Benzylpenicillin Potassium disk method under Infrared Spectrophotometry <2.25>, and
(C16H17KN2O4S) compare the spectrum with the Reference Spectrum: both
＝ M S × AT / AS spectra exhibit similar intensities of absorption at the same
wave numbers.
MS: Amount (unit) of Benzylpenicillin Potassium RS
(3) Perform the test with Bepotastine Besilate as directed
taken
under Flame Coloration Test <1.04> (2): a green color ap-
Operating conditions— pears.
Detector: An ultraviolet absorption photometer (wave- (4) Mix well 30 mg of Bepotastine Besilate with 0.1 g of
length: 254 nm). sodium nitrate and 0.1 g of anhydrous sodium carbonate,
Column: A stainless steel column 4.6 mm in inside diame- and gradually ignite. After cooling, dissolve the residue in 2
ter and 25 cm in length, packed with octadecylsilanized silica mL of dilute hydrochloric acid and 10 mL of water, filter if
gel for liquid chromatography (7 mm in particle diameter). necessary, and add barium chloride TS: a white precipitate is
Column temperature: A constant temperature of about produced.
259 C.
Melting point <2.60> 159–1639C
Mobile phase: To a mixture of diammonium hydrogen
phosphate solution (33 in 5000) and acetonitril (19:6), add Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
phosphoric acid to adjust the pH of this solution to 8.0. Bepotastine Besilate according to Method 4, and perform the
Flow rate: Adjust so that the retention time of benzyl- test. Prepare the control solution with 2.0 mL of Standard
penicillin is about 7.5 minutes. Lead Solution (not more than 10 ppm).
System suitability— (2) Related substances—Dissolve 10 mg of Bepotastine
System performance: When the procedure is run with 5 mL Besilate in 25 mL of the mobile phase, and use this solution
of the standard solution under the above operating condi- as the sample solution. Pipet 1 mL of the sample solution,
tions, the number of theoretical plates and the symmetry fac- add the mobile phase to make exactly 100 mL, and use this
tor of the peak of benzylpenicillin are not less than 6000 and solution as the standard solution. Perform the test with
not more than 2.0, respectively. exactly 20 mL each of the sample solution and standard
System repeatability: When the test is repeated 6 times solution as directed under Liquid Chromatography <2.01>
with 5 mL of the standard solution under the above operating according to the following conditions, and determine each
JP XVII Official Monographs / Bepotastine Besilate Tablets 489
peak area by the automatic integration method: the area of drogen phosphate TS and acetonitrile (3:1).
the peak, having a relative retention time of about 2.5 to Flow rate: Adjust so that the retention time of bepotastine
bepotastine, obtained from the sample solution is not larger is about 17 minutes.
than the peak area of bepotastine obtained from the stand- System suitability—
ard solution, and the area of the peak other than bepotastine System performance: When the procedure is run with 10
and the peak mentioned above from the sample solution is mL of the standard solution under the above operating con-
not larger than 1/10 times the peak area of bepotastine from ditions, the number of theoretical plates and the symmetry
the standard solution. Furthermore, the total area of the factor of the peak of bepotastine are not less than 3000 and
peaks other than bepotastine from the sample solution is not 0.8 to 1.5, respectively.
larger than the peak area of bepotastine from the standard System repeatability: When the test is repeated 6 times
solution. with 10 mL of the standard solution under the above operat-
Operating conditions— ing conditions, the relative standard deviation of the peak
Detector: An ultraviolet absorption photometer (wave- area of bepotastine is not more than 5.0z.
length: 220 nm).
Water <2.48> Not more than 0.1z (0.3 g, coulometric
titration).
ter and 15 cm in length, packed with octylsilanized
silica gel for liquid chromatography (5 mm in particle diame- Residue on ignition <2.44> Not more than 0.1z (1 g).
ter).
Assay Weigh accurately about 0.8 g of Bepotastine Besi-
late, dissolve in 60 mL of acetic acid (100), and titrate <2.50>
409 C.
with 0.1 mol/L perchloric acid VS (potentiometric titration).
Mobile phase: Dissolve 1.0 g of sodium 1-pentane sul-
Perform a blank determination in the same manner, and
fonate in a mixture of 0.05 mol/L potassium dihydrogen
make any necessary correction.
phosphate TS (pH 3.0) and acetonitrile (7:3) to make 1000
mL. Each mL of 0.1 mol/L perchloric acid VS
Flow rate: Adjust so that the retention time of bepotastine ＝ 54.71 mg of C21H25ClN2O3.C6H6O3S
is about 6 minutes.
retention time of bepotastine, beginning after the peak of
benzenesulfonic acid.
System suitability— Bepotastine Besilate Tablets
Test for required detectability: Pipet 2.5 mL of the stand-
ベポタスチンベシル酸塩錠
ard solution, and add the mobile phase to make exactly 50
mL. Confirm that the peak area of bepotastine obtained
with 20 mL of this solution is equivalent to 3.5 to 6.5z of Bepotastine Besilate Tablets contain not less than
that obtained with 20 mL of the standard solution. 95.0z and not more than 105.0z of the labeled amount
System performance: When the procedure is run with 20 of bepotastine besilate (C21H25ClN2O3.C6H6O3S:
mL of the standard solution under the above operating con- 547.06).
factor of the peak of bepotastine are not less than 3000 and
with Bepotastine Besilate.
0.8 to 1.5, respectively.
System repeatability: When the test is repeated 6 times Identification To an amount of powdered Bepotastine
with 20 mL of the standard solution under the above operat- Besilate Tablets, equivalent to 2 mg of Bepotastine Besilate,
ing conditions, the relative standard deviation of the peak add 40 mL of water, shake thoroughly, and filter. Determine
area of bepotastine is not more than 2.0z. the absorption spectrum of the filtrate as directed under
(3) Optical isomer—Dissolve 5.0 mg of Bepotastine Besi- Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a
late in 25 mL of the mobile phase, and use this solution as maximum between 260 nm and 264 nm.
the mobile phase to make exactly 200 mL, and use this solu-
To 1 tablet of Bepotastine Besilate Tablets add exactly
V/5 mL of the internal standard solution, then add the mo-
the following conditions, and determine the area of each
bile phase to make V mL so that each mL contains about 0.4
peak by the automatic integration method: the area of the
mg of bepotastine besilate (C21H25ClN2O3.C6H6O3S), shake
peak, having a relative retention time of about 0.9 to
vigorously for 10 minutes, and filter through a membrane
bepotastine obtained from the sample solution, is not larger
than the peak area of bepotastine obtained from the stand-
first 5 mL of the filtrate, to 2 mL of the subsequent filtrate
ard solution.
add the mobile phase to make 10 mL, and use this solution
as the sample solution. Then, proceed as directed in the
Assay.
length: 225 nm).
Column: A stainless steel column 6.0 mm in inside diame- Amount (mg) of bepotastine besilate
ter and 15 cm in length, packed with b-cyclodextrin binding (C21H25ClN2O3.C6H6O3S)
silica gel for liquid chromatography (5 mm in particle diame- ＝ MS × QT/QS × V/50
ter).
MS: Amount (mg) of bepotastine besilate for assay taken,
calculated on the anhydrous and residual solvent-free
409 C.
basis
490 Beraprost Sodium / Official Monographs JP XVII
Internal standard solution—A solution of ethyl parahy- as the standard solution. Perform the test with 20 mL each of
droxybenzoate in acetonitrile (1 in 4500). the sample solution and standard solution as directed under
area of bepotastine to that of the internal standard.
in 30 minutes of Bepotastine Besilate Tablets is not less than Amount (mg) of bepotastine besilate
85z. (C21H25ClN2O3.C6H6O3S)
Start the test with 1 tablet of Bepotastine Besilate Tablets, ＝ MS × QT/QS × 1/2
withdraw not less than 20 mL of the medium at the specified
MS: Amount (mg) of bepotastine besilate for assay taken,
calculated on the anhydrous and residual solvent-free
basis
filtrate, add the mobile phase to make exactly V? mL so that Internal standard solution—A solution of ethyl parahy-
each mL contains about 2.2 mg of bepotastine besilate droxybenzoate in acetonitrile (1 in 4500).
(C21H25ClN2O3.C6H6O3S), and use this solution as the sam- Operating conditions—
ple solution. Separately, weigh accurately about 0.11 g of Detector: An ultraviolet absorption photometer (wave-
bepotastine besilate for assay (separately determine the water length: 260 nm).
<2.48> and the residual solvent in the same manner as Column: A stainless steel column 4.6 mm in inside diame-
Bepotastine Besilate), and dissolve in water to make exactly ter and 15 cm in length, packed with octylsilanized silica gel
100 mL. Pipet 1 mL of this solution, add water to make for liquid chromatography (5 mm in particle diameter).
exactly 100 mL. Pipet 2 mL of this solution, add the mobile Column temperature: A constant temperature of about
phase to make exactly 10 mL, and use this solution as the 409C.
standard solution. Perform the test with exactly 50 mL each Mobile phase: A solution of sodium 1-pentanesulfonate in
of the sample solution and standard solution as directed a mixture of 0.05 mol/L potassium dihydrogen phosphate
under Liquid Chromatography <2.01> according to the fol- TS (pH 3.0) and acetonitrile (7:3) (1 in 1000).
lowing conditions, and determine the peak areas, AT and AS, Flow rate: Adjust so that the retention time of bepotastine
of bepotastine in each solution. is about 6 minutes.
of bepotastine besilate (C21H25ClN2O3.C6H6O3S)
＝ MS × AT/AS × V?/V × 1/C × 9/5
ditions, bepotastine and the internal standard are eluted in
MS: Amount (mg) of bepotastine besilate for assay taken, this order with the resolution between these peaks being not
calculated on the anhydrous and residual solvent-free less than 5.
basis System repeatability: When the test is repeated 6 times
C: Labeled amount (mg) of bepotastine besilate with 20 mL of the standard solution under the above operat-
(C21H25ClN2O3.C6H6O3S) in 1 tablet ing conditions, the relative standard deviation of the ratio of
the peak area of bepotastine to that of the internal standard
Assay. Containers and storage Containers—Tight containers.
mL of the standard solution under the above operating con- Beraprost Sodium
factor of the peak of bepotastine are not less than 5000 and ベラプロストナトリウム
area of bepotastine is not more than 1.5z.
Assay Weigh accurately the mass of not less than 20 tablets
of Bepotastine Besilate Tablets, and powder. Weigh accu-
rately a portion of the powder, equivalent to about 10 mg of
bepotastine besilate (C21H25ClN2O3.C6H6O3S), add exactly 5
mL of the internal standard solution, then add 20 mL of the
mobile phase, shake thoroughly for 10 minutes, and filter C24H29NaO5: 420.47
through a membrane filter with a pore size not exceeding Monosodium (1RS,2RS,3aSR,8bSR )-2,3,3a,8b-tetrahydro-2-hydroxy-
0.45 mm. Discard the first 5 mL of the filtrate, to 2 mL of the 1-[(1E,3SR,4RS )-3-hydroxy-4-methyloct-1-en-6-yn-1-yl]-1H-
subsequent filtrate add the mobile phase to make 10 mL, and cyclopenta[b]benzofuran-5-butanoate
use this solution as the sample solution. Separately, weigh Monosodium (1RS,2RS,3aSR,8bSR )-2,3,3a,8b-tetrahydro-2-hydroxy-
accurately about 20 mg of bepotastine besilate for assay 1-[(1E,3SR,4SR )-3-hydroxy-4-methyloct-1-en-6-yn-1-yl]-1H-
(separately determine the water <2.48> and the residual sol- cyclopenta[b]benzofuran-5-butanoate
vent in the same manner as Bepotastine Besilate), add ex- [88475-69-8]
actly 10 mL of the internal standard solution, and dissolve in
the mobile phase to make 50 mL. To 2 mL of this solution Beraprost Sodium, when dried, contains not less
add the mobile phase to make 10 mL, and use this solution than 98.5z and not more than 101.0z of beraprost
JP XVII Official Monographs / Beraprost Sodium 491
sodium (C24H29NaO5). peak of beraprost is about 23 minutes.

Time span of measurement: For 80 minutes after injec-
Description Beraprost Sodium occurs as a white powder.
tion, beginning after the solvent peak.
It is very soluble in methanol, and freely soluble in water
and in ethanol (99.5).
It is hygroscopic.
lution add methanol to make 20 mL. To 1 mL of this solu-
A solution of Beraprost Sodium (1 in 200) shows no opti-
tion add methanol to make 20 mL, and use this solution as
cal rotation.
the solution for system suitability test. Pipet 2 mL of the
Identification (1) Determine the absorption spectrum of a solution for system suitability test, and add methanol to
solution of Beraprost Sodium in methanol (3 in 50,000) as make exactly 10 mL. Confirm that the total area of the two
directed under Ultraviolet-visible Spectrophotometry <2.24>, peaks of beraprost obtained with 15 mL of this solution is
and compare the spectrum with the Reference Spectrum: equivalent to 14 to 26z of that with 15 mL of the solution
both spectra exhibit similar intensities of absorption at the for system suitability test.
same wavelengths. System performance: When the procedure is run with 15
(2) Determine the infrared absorption spectrum of previ- mL of the solution for system suitability test under the above
ously dried Beraprost Sodium as directed in the potassium operating conditions, the resolution between the two peaks
bromide disk method under Infrared Spectrophotometry of beraprost is not less than 1.5.
<2.25>, and compare the spectrum with the Reference Spec- System repeatability: When the test is repeated 6 times
trum: both spectra exhibit similar intensities of absorption at with 15 mL of the solution for system suitability test under
the same wave numbers. the above operating conditions, the relative standard devia-
(3) A solution of Beraprost Sodium in methanol (1 in tion of the total area of the two peaks of beraprost is not
1000) responds to the Qualitative Tests <1.09> (1) for sodium more than 2.0z.
salt.
Purity Related substances—Dissolve 20 mg of Beraprost pressure not exceeding 0.67 kPa, silica gel, 609C, 5 hours).
Sodium in 2 mL of methanol, and use this solution as the
Isomer ratio Dissolve 10 mg of Beraprost Sodium in 5 mL
sample solution. Perform the test with 15 mL of the sample
of methanol, and use this solution as the sample solution.
Perform the test with 15 mL of the sample solution as di-
area by the automatic integration method, and calculate
the following conditions, and determine the areas, Aa of the
their amounts by the area percentage method: the amount of
peak which appears at the retention time about 25 minutes,
the peak having the relative retention time of about 0.5 to
and Ab of the peak which appears at about 27 minutes: Ab/
the second eluting principal peak of beraprost and the adja-
Aa is between 0.90 and 1.10.
cent two peaks having the relative retention time of about
1.7 and another adjacent two peaks having the relative reten-
tion time of about 2.0 are not more than 0.2z, respectively,
length: 285 nm).
the amount of the peak having the relative retention time of
about 1.2 is not more than 0.3z, the amount of the peak,
other than the two peaks of beraprost and the peaks men-
tioned above, is less than 0.1z, and the total amount of the
peaks, other than the two peaks of beraprost, is not more
409C.
than 1.5z.
Mobile phase: A mixture of methanol, water and acetic
acid (100) (600:400:1).
Flow rate: Adjust so that the retention time of the second
length: 285 nm).
eluting peak of beraprost is about 27 minutes.
mL of the sample solution under the above operating condi-
tions, the resolution between the two peaks of beraprost is
359 C.
not less than 1.2.
Mobile phase A: A mixture of water, acetonitrile, metha-
nol and acetic acid (100) (640:330:30:1).
with 15 mL of the sample solution under the above operating
Mobile phase B: A mixture of acetonitrile, water and
conditions, the relative standard deviation of the total area
acetic acid (100) (900:100:1).
of the two peak of beraprost is not more than 2.0z.
the mobile phases A and B as directed in the following table. Assay Weigh accurately about 0.1 g of Beraprost Sodium,
previously dried, dissolve in 30 mL of diluted ethanol with
Time after injection Mobile phase A Mobile phase B freshly boiled and cooled water (7 in 10), add exactly 2 mL
of sample (min) (volz) (volz) of 0.2 mol/L hydrochloric acid TS, and titrate <2.50> with
0.025 mol/L sodium hydroxide-ethanol VS from the first
0 – 30 100 0 equivalence point to the second equivalence point (potentio-
30 – 45 100 → 56 0 → 44 metric titration).
45 – 60 56 44
Each mL of 0.025 mol/L sodium hydroxide-ethanol VS
60 – 70 56 → 0 44 → 100
＝ 10.51 mg of C24H29NaO5
70 – 80 0 100
Flow rate: Adjust so that the retention time of the second Storage—Light-resistant.
492 Beraprost Sodium Tablets / Official Monographs JP XVII
solve in methanol to make exactly 100 mL. Pipet 2 mL of
Beraprost Sodium Tablets this solution, and add water to make exactly 200 mL. Pipet 2
ベラプロストナトリウム錠 use this solution as the standard solution. Perform the test
Beraprost Sodium Tablets contain not less than
total areas, AT and AS, of the two peaks of beraprost in each
amount of beraprost sodium (C24H29NaO5: 420.47).
solution.
with Beraprost Sodium.
of beraprost sodium (C24H29NaO5)
Identification Powder Beraprost Sodium Tablets. To a ＝ MS × AT/AS × V?/V × 1/C × 9/100
portion of the powder, equivalent to 0.2 mg of Beraprost
MS: Amount (mg) of beraprost sodium for assay taken
Sodium, add 10 mL of water, shake, and filter through a
C: Labeled amount (mg) of beraprost sodium
membrane filter with a pore size not exceeding 0.45 mm. To
(C24H29NaO5) in 1 tablet
the filtrate add 1 mL of 0.1 mol/L hydrochloric acid TS, ex-
tract with two 50-mL portions of ethyl acetate, combine the Operating conditions—
extracts, and evaporate in reduced pressure at 409C. Dis- Detector, column temperature, and mobile phase: Proceed
solve the residue in 1 mL of methanol, use this solution as as directed in the operating conditions in the Assay.
the sample solution. Separately, dissolve 1 mg of beraprost Column: A stainless steel column 4.6 mm in inside diame-
sodium in 5 mL of methanol, and use this solution as the ter and 15 cm in length, packed with octadecylsilanized silica
standard solution. Perform the test with these solutions as gel for liquid chromatography (5 mm in particle diameter).
directed under Thin-layer Chromatography <2.03>. Spot 10 Flow rate: Adjust so that the retention time of the first
mL each of the sample solution and standard solution on a eluting peak of beraprost is about 10 minutes.
plate of silica gel for thin-layer chromatography, develop the System suitability—
plate with the upper layer of a mixture of 11 volumes of System performance: When the procedure is run with 200
ethyl acetate, 10 volumes of water, 4 volumes of isooctane mL of the standard solution under the above operating con-
and 2 volumes of acetic acid (100) to a distance of about 10 ditions, the resolution between the two peaks of beraprost is
cm, air-dry the plate, and heat at 1209 C for 30 minutes. not less than 1.2.
After cooling, spray evenly a mixture of ethanol (99.5), System repeatability: When the test is repeated 6 times
water, sulfuric acid and 4-methoxybenzaldehide (17:2:1:1) with 200 mL of the standard solution under the above operat-
on the plate, and heat at 1209C for 3 minutes: the principal ing conditions, the relative standard deviation of the total
spot obtained from the sample solution and the spot ob- area of the two peaks of beraprost is not more than 2.0z.
tained from the standard solution show the same R f value.
Assay Weigh accurately the mass of not less than 20 Ber-
Uniformity of dosage units <6.02> Perform the test accord- aprost Sodium Tablets, and powder. Weigh accurately a por-
ing to the following method: it meets the requirement of the tion of the powder, equivalent to about 40 mg of beraprost
Content uniformity test. sodium (C24H29NaO5), add exactly 20 mL of the internal
To 1 tablet of Beraprost Sodium Tablets add exactly V mL standard solution, shake at 309C for 30 minutes, filter
of the internal standard solution so that each mL contains through a membrane filter with a pore size not exceeding
about 2 mg of beraprost sodium (C24H29NaO5), shake at 0.45 mm, and use the filtrate as the sample solution. Sepa-
309 C for 30 minutes, filter through a membrane filter with a rately, weigh accurately about 20 mg of beraprost sodium
pore size not exceeding 0.45 mm, and use the filtrate as the for assay, previously dried in reduced pressure not exceeding
sample solution. Then, proceed as directed in the Assay. 0.67 kPa at 609C for 5 hours using silica gel as a desiccant,
and dissolve in methanol to make exactly 100 mL. Pipet
Amount (mg) of beraprost sodium (C24H29NaO5)
10 mL of this solution, and add methanol to make exactly
＝ MS × QT/QS × V/10,000
200 mL. Pipet 4 mL of this solution, and evaporate under
MS: Amount (mg) of beraprost sodium for assay taken reduced pressure at 409 C. To the residue add exactly 20 mL
of the internal standard solution, and use this solution as the
Internal standard solution—A mixture of water and a solu-
tion of 4-isopropylphenol in methanol (1 in 250,000) (1:1).
Dissolution <6.10> When the test is performed at 50 revolu- uid Chromatography <2.01> according to the following con-
tions per minute according to the Paddle method, using 900 ditions, and calculate the ratios, QT and QS, of the total area
mL of water as the dissolution medium, the dissolution rate of the two peaks of beraprost to the peak area of the internal
in 30 minutes of Beraprost Sodium Tablets is not less than standard.
85z.
Amount (mg) of beraprost sodium (C24H29NaO5)
Start the test with 1 tablet of Beraprost Sodium Tablets,
＝ MS × QT/QS × 1/500
minute after starting the test, and filter through a membrane MS: Amount (mg) of beraprost sodium for assay taken
Internal standard solution—A mixture of water and a solu-
tion of 4-isopropylphenol in methanol (1 in 250,000) (1:1).
trate, add water to make exactly V? mL so that each mL
contains about 22 ng of beraprost sodium (C24H29NaO5),
Detector: A fluorophotometer (excitation wavelength:
285 nm, fluorescence wavelength: 614 nm).
weigh accurately about 20 mg of beraprost sodium for assay,
previously dried in reduced pressure not exceeding 0.67 kPa
at 609C for 5 hours using silica gel as a desiccant, and dis-
JP XVII Official Monographs / Berberine Chloride Hydrate 493
for liquid chromatography (5 mm in particle diameter). mL of water by warming, add 0.5 mL of nitric acid, cool,
Column temperature: A constant temperature of about and filter after allowing to stand for 10 minutes. To 3 mL of
409 C. the filtrate add 1 mL of silver nitrate TS, and collect the pro-
Mobile phase: A mixture of methanol, water and acetic duced precipitate: the precipitate does not dissolve in dilute
acid (100) (650:350:1). nitric acid, but it dissolves in an excess amount of ammonia
Flow rate: Adjust so that the retention time of the first TS.
eluting peak of beraprost is about 15 minutes.
Purity (1) Acidity—Shake thoroughly 0.10 g of Berberine
Chloride Hydrate with 30 mL of water, and filter. To the fil-
trate add 2 drops of phenolphthalein TS and 0.10 mL of 0.1
mol/L sodium hydroxide VS: the yellow color changes to an
ditions, the internal standard and beraprost are eluted in this
orange to red color.
order and the resolution between the internal standard peak
(2) Sulfate <1.14>—Shake 1.0 g of Berberine Chloride
and the first eluting peak of beraprost is not less than 11,
Hydrate with 48 mL of water and 2 mL of dilute hydrochlo-
and the resolution between the two peaks of beraprost is not
ric acid for 1 minute, and filter. Discard the first 5 mL of the
less than 1.5.
filtrate, take the subsequent 25 mL of the filtrate, add water
the test solution. Prepare the control solution with 0.50 mL
of 0.005 mol/L sulfuric acid VS, 1 mL of dilute hydrochloric
the total area of the two peaks of beraprost to the peak area
acid, 5 to 10 drops of bromophenol blue TS and water to
of the internal standard is not more than 2.0z.
Containers and storage Containers—Well-closed contain- (3) Heavy metals <1.07>—Proceed with 1.0 g of Berber-
ers. ine Chloride Hydrate according to Method 2, and perform
Berberine Chloride Hydrate (4) Related substances—Dissolve 10 mg of Berberine
Chloride Hydrate in 100 mL of the mobile phase, and use
ベルベリン塩化物水和物 this solution as the sample solution. Pipet 4 mL of the sam-
ple solution, add the mobile phase to make exactly 100 mL,
test with exactly 10 mL each of the sample solution and
<2.01> according to the following conditions. Determine each
method: the total area of the peaks other than berberine ob-
tained with the sample solution is not larger than the peak
area of berberine obtained with the standard solution.
C20H18ClNO4.xH2O
9,10-Dimethoxy-5,6-
Detector, column, column temperature, mobile phase,
dihydro[1,3]dioxolo[4,5-g]isoquino[3,2-a]isoquinolin-
flow rate, and selection of column: Proceed as directed in
7-ium chloride hydrate
the operating conditions in the Assay.
[633-65-8, anhydride]
retention time of berberine, beginning after the solvent peak.
Berberine Chloride Hydrate contains not less than
Detection sensitivity: Adjust so that the peak height of
95.0z and not more than 102.0z of berberine chlo-
berberine obtained from 10 mL of the standard solution is
ride (C20H18ClNO4: 371.81), calculated on the anhy-
about 10z of the full scale.
drous basis.
Water <2.48> 8 – 12z (0.1 g, volumetric titration, direct
Description Berberine Chloride Hydrate occurs as yellow,
titration).
crystals or crystalline powder. It is odorless or has a faint,
characteristic odor. It has a very bitter taste. Residue on ignition <2.44> Not more than 0.1z (1 g).
It is sparingly soluble in methanol, slightly soluble in
Assay Weigh accurately about 10 mg of Berberine Chloride
ethanol (95), and very slightly soluble in water.
Hydrate, dissolve in the mobile phase to make exactly 100
Identification (1) Determine the absorption spectrum of a mL, and use this solution as the sample solution. Separately,
solution of Berberine Chloride Hydrate (1 in 100,000) as weigh accurately about 10 mg of Berberine Chloride RS
directed under Ultraviolet-visible Spectrophotometry <2.24>, (separately, determine the water content <2.48> in the same
and compare the spectrum with the Reference Spectrum manner as Berberine Chloride Hydrate), and dissolve in the
or the spectrum of a solution of Berberine Chloride RS mobile phase to make exactly 100 mL, and use this solution
prepared in the same manner as the sample solution: both as the standard solution. Perform the test with exactly 10 mL
spectra exhibit similar intensities of absorption at the same each of the sample solution and standard solution as directed
wavelengths. under Liquid Chromatography <2.01> according to the fol-
(2) Determine the infrared absorption spectrum of Ber- lowing conditions. Determine the peak areas, AT and AS of
berine Chloride Hydrate as directed in the potassium bro- berberine in each solution.
Amount (mg) of berberine chloride (C20H18ClNO4)
＝ M S × AT / AS
the spectrum of Berberine Chloride RS: both spectra exhibit
similar intensities of absorption at the same wave numbers. MS: Amount (mg) of Berberine Chloride RS taken, calcu-
(3) Dissolve 0.1 g of Berberine Chloride Hydrate in 20 lated on the anhydrous basis
494 Berberine Tannate / Official Monographs JP XVII
Operating conditions— 30 mL of water, and filter after shaking well. To the filtrate
Detector: An ultraviolet absorption photometer (wave- add 2 drops of phenolphthalein TS and 0.10 mL of 0.1
length: 345 nm). mol/L sodium hydroxide VS: the color of the solution
Column: A stainless steel column about 4 mm in inside changes from yellow to orange to red.
diameter and about 25 cm in length, packed with octadecyl- (2) Chloride <1.03>—Shake 1.0 g of Berberine Tannate
silanized silica gel for liquid chromatography (5 mm in parti- with 38 mL of water and 12 mL of dilute nitric acid for 5
cle diameter). minutes, and filter. Discard the first 5 mL of the filtrate, to
Column temperature: A constant temperature of about 25 mL of the subsequent filtrate add water to make 50 mL,
409 C. and perform the test using this solution as the test solution.
Mobile phase: Dissolve 3.4 g of monobasic potassium Prepare the control solution with 0.50 mL of 0.01 mol/L hy-
phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a drochloric acid VS by adding 6 mL of dilute nitric acid, 10 to
mixture of water and acetonitrile (1:1). 15 drops of bromophenol blue TS and water to make 50 mL
Flow rate: Adjust so that the retention time of berberine is (not more than 0.035z).
about 10 minutes. (3) Sulfate <1.14>—Shake 1.0 g of Berberine Tannate
Selection of column: Dissolve each 1 mg of berberine with 48 mL of water and 2 mL of dilute hydrochloric acid
chloride and palmatin chloride in the mobile phase to make for 1 minute, and filter. Discard the first 5 mL of the filtrate,
10 mL. Proceed with 10 mL of this solution under the above take the subsequent 25 mL of the filtrate, add water to make
operating conditions, and calculate the resolution. Use a 50 mL, and perform the test using this solution as the test
column giving elution of palmatin and berberine in this solution. Prepare the control solution with 0.50 mL of 0.005
order with the resolution between these peaks being not less mol/L sulfuric acid VS, 1 mL of dilute hydrochloric acid, 5
than 1.5. to 10 drops of bromophenol blue TS and water to make 50
System repeatability: When the test is repeated 5 times mL (not more than 0.048z).
with the standard solution under the above operating condi- (4) Heavy metals <1.07>—Proceed with 1.0 g of Berber-
tions, the relative standard deviation of the peak areas of ine Tannate according to Method 2, and perform the test.
berberine is not more than 1.5z. Prepare the control solution with 3.0 mL of Standard Lead
(5) Related substances—Dissolve 10 mg of Berberine
Tannate in 100 mL of the mobile phase, and use this solution
Berberine Tannate solution as the standard solution. Perform the test with
タンニン酸ベルベリン
Berberine Tannate is a compound of berberine and area of both solutions by the automatic integration method:
tannic acid. the total area of the peaks other than berberine obtained
It contains not less than 27.0z and not more than with the sample solution is not larger than the peak area of
33.0z of berberine (C20H19NO5: 353.37), calculated berberine obtained with the standard solution.
on the anhydrous basis. Operating conditions—
Description Berberine Tannate occurs as a yellow to light
yellow-brown powder. It is odorless or has a faint, charac-
the Assay.
teristic odor, and is tasteless.
It is practically insoluble in water, in acetonitrile, in meth-
retention time of berberine, beginning after the solvent peak.
anol and in ethanol (95).
Identification (1) To 0.1 g of Berberine Tannate add 10 System performance: Proceed as directed in the system
mL of ethanol (95), and heat in a water bath for 3 minutes suitability in the Assay.
with shaking. Cool, filter, and to 5 mL of the filtrate add 1 Test for required detectability: To exactly 2 mL of the
drop of iron (III) chloride TS: a blue-green color is pro- standard solution add the mobile phase to make exactly 20
duced, and on allowing to stand, a bluish black precipitate is mL. Confirm that the peak area of berberine obtained with
formed. 10 mL of this solution is equivalent to 7 to 13z of that ob-
(2) Dissolve 0.01 g of Berberine Tannate in 10 mL of tained with 10 mL of the standard solution.
methanol and 0.4 mL of 1 mol/L hydrochloric acid TS, and System repeatability: When the test is repeated 6 times
add water to make 200 mL. To 8 mL of the solution add with 10 mL of the standard solution under the above operat-
water to make 25 mL. Determine the absorption spectrum of ing conditions, the relative standard deviation of the peak
the solution as directed under Ultraviolet-visible Spectropho- area of berberine is not more than 3.0z.
(3) Determine the infrared absorption spectrum of Ber- Residue on ignition <2.44> Not more than 1.0z (1 g).
berine Tannate as directed in the potassium bromide disk
Assay Weigh accurately about 30 mg of Berberine Tan-
nate, dissolve in the mobile phase to make exactly 100 mL,
weigh accurately about 10 mg of Berberine Chloride RS
numbers.
(separately, determine the water <2.48> in the same manner
Purity (1) Acidity—To 0.10 g of Berberine Tannate add as Berberine Chloride Hydrate), dissolve in the mobile phase
JP XVII Official Monographs / Betahistine Mesilate 495
to make exactly 100 mL, and use this solution as the stand- Reference Spectrum: both spectra exhibit similar intensities
ard solution. Perform the test with exactly 10 mL each of the of absorption at the same wavelengths.
sample solution and standard solution as directed under Liq- (2) Determine the infrared absorption spectrum of Beta-
uid Chromatography <2.01> according to the following con- histine Mesilate, previously dried, as directed in the potas-
ditions. Determine the peak areas, AT and AS, of berberine sium bromide disk method under Infrared Spectrophotome-
in each solution. try <2.25>, and compare the spectrum with the Reference
Amount (mg) of berberine (C20H19NO5)
＝ MS × AT/AS × 0.950
(3) A 30 mg portion of Betahistine Mesilate responds to
MS: Amount (mg) of Berberine Chloride RS taken, calcu- the Qualitative Tests <1.09> (2) for mesilate.
lated on the anhydrous basis
Melting point <2.60> 110 – 1149C (after drying).
Betahistine Mesilate according to Method 4, and perform the
length: 345 nm).
(2) Related substances—Dissolve 50 mg of Betahistine
Mesilate in 10 mL of a mixture of water and acetonitrile
409 C.
mL of the sample solution, add the mixture of water and
acetonitrile (63:37) to make exactly 100 mL, and use this so-
phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a
lution as the standard solution. Perform the test with exactly
mixture of water and acetonitrile (1:1).
Flow rate: Adjust so that the retention time of berberine is
about 10 minutes.
the automatic integration method: the area of the peak other
System performance: Dissolve 1 mg each of berberine
than betahistine with the sample solution is not larger than
chloride and palmatin chloride in the mobile phase to make
1/10 times the peak area of betahistine with the standard so-
10 mL. When the procedure is run with 10 mL of this solu-
lution, and the total area of the peaks other than the peak of
tion under the above operating conditions, palmatin and ber-
betahistine with the sample solution is not larger than 1/2
berine are eluted in this order with the resolution between
times the peak area of betahistine with the standard solution.
these peaks being not less than 1.5.
length: 261 nm).
area of berberine is not more than 1.5z.
Containers and storage Containers—Tight containers. gel for liquid chromatography (5 mm in particle diameter).
Storage—Light-resistant. Column temperature: A constant temperature of about
359C.
Mobile phase: To 5 mL of diethylamine and 20 mL of
Betahistine Mesilate acetic acid (100) add water to make 1000 mL. Dissolve 2.3 g
of sodium lauryl sulfate in 630 mL of this solution, and add
ベタヒスチンメシル酸塩 370 mL of acetonitrile.
Flow rate: Adjust so that the retention time of betahistine
is about 5 minutes.
retention time of betahistine, beginning after the solvent
peak.
C8H12N2.2CH4O3S: 328.41
N-Methyl-2-pyridin-2-ylethylamine
dimethanesulfonate
standard solution add the mixture of water and acetonitrile
[5638-76-6, Betahistine]
(63:37) to make exactly 50 mL. Confirm that the peak area
of betahistine obtained with 20 mL of this solution is equiva-
Betahistine Mesilate, when dried, contains not less
lent to 7 to 13z of that obtained with 20 mL of the standard
than 98.0z and not more than 101.0z of betahistine
solution.
mesilate (C8H12N2.2CH4O3S).
System performance: Dissolve 10 mg of betahistine mesi-
Description Betahistine Mesilate occurs as white, crystals late and 10 mg of 2-vinylpyridine in 50 mL of the mixture of
or crystalline powder. water and acetonitrile (63:37). To 2 mL of this solution add
It is very soluble in water, freely soluble in acetic acid the mixture of water and acetonitrile (63:37) to make 50 mL.
(100), and sparingly soluble in ethanol (99.5). When the procedure is run with 20 mL of this solution under
It dissolves in dilute hydrochloric acid. the above operating conditions, 2-vinylpyridine and beta-
It is hygroscopic. histine are eluted in this order with the resolution between
solution of Betahistine Mesilate in 0.1 mol/L hydrochloric
acid (1 in 50,000) as directed under Ultraviolet-visible Spec-
area of betahistine is not more than 1.0z
496 Betahistine Mesilate Tablets / Official Monographs JP XVII
Loss on drying <2.41> Not more than 1.0z (1 g, in vacu- late and 10 mg of 2-vinylpyridine in 50 mL of the mixture of
um, phosphorus (V) oxide, 709C, 24 hours). water and acetonitrile (63:37). To 2 mL of this solution add
the mixture of water and acetonitrile (63:37) to make 50 mL.
When the procedure is run with 20 mL of this solution under
Assay Weigh accurately about 0.2 g of Betahistine Mesi- the above operating conditions, 2-vinylpyridine and beta-
late, previously dried, dissolve in 1 mL of acetic acid (100), histine are eluted in this order with the resolution between
add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 these peaks being not less than 5.
mol/L perchloric acid VS (potentiometric titration). Per- System repeatability: When the test is repeated 6 times
form a blank determination, and make any necessary correc- with 20 mL of the standard solution under the above operat-
tion. ing conditions, the relative standard deviation of the peak
area of betahistine is not more than 1.0z.
＝ 16.42 mg of C8H12N2.2CH4O3S Uniformity of dosage units <6.02> Perform the test accord-
To 1 tablet of Betahistine Mesilate Tablets add exactly
V mL of 0.1 mol/L hydrochloric acid TS so that each
Betahistine Mesilate Tablets mL contains about 0.4 mg of betahistine mesilate
(C8H12N2.2CH4O3S), agitate for about 10 minutes with the
ベタヒスチンメシル酸塩錠
aid of ultrasonic waves to disintegrate the tablet, then centri-
fuge, and use the supernatant liquid as the sample solution.
Betahistine Mesilate Tablets contain not less than Proceed as directed in the Assay.
Amount (mg) of betahistine mesilate (C8H12N2.2CH4O3S)
amount of betahistine mesilate (C8H12N2.2CH4O3S: ＝ MS × AT/AS × V/250
328.41).
MS: Amount (mg) of betahistine mesilate for assay taken
with Betahistine Mesilate. Dissolution <6.10> When the test is performed at 50 revolu-
Identification To 5 mL of the sample solution obtained in
the Assay add 0.1 mol/L hydrochloric acid TS to make 100
in 15 minutes of Betahistine Mesilate Tablets is not less than
mL, and determine the absorption spectrum of this solution
85z.
Start the test with 1 tablet of Betahistine Mesilate Tablets,
Purity Related substances—Powder not less than 20 Beta- minute after starting the test, and filter through a membrane
histine Mesilate Tablets. To a portion of the powder, equiva- filter with a pore size not exceeding 0.45 mm. Discard the
lent to about 50 mg of Betahistine Mesilate, add 10 mL of a first 10 mL of the filtrate, pipet V mL of the subsequent
mixture of water and acetonitrile (63:37), agitate for 10 filtrate, add water to make exactly V? mL so that each mL
minutes with the aid of ultrasonic waves, centrifuge, and use contains about 6.7 mg of betahistine mesilate (C8H12N2.
the supernatant liquid as the sample solution. Pipet 1 mL of 2CH4O3S), and use this solution as the sample solution.
the sample solution, add the mixture of water and aceto- Separately, weigh accurately about 17 mg of betahistine
nitrile (63:37) to make exactly 100 mL, and use this solution mesilate for assay, previously dried under reduced pressure
as the standard solution. Perform the test with exactly 20 mL with phosphorous (V) oxide at 709 C for 24 hours, and dis-
each of the sample solution and standard solution as directed solve in water to make exactly 100 mL. Pipet 4 mL of this
under Liquid Chromatography <2.01> according to the fol- solution, add water to make exactly 100 mL, and use this so-
lowing conditions, and determine each peak area by the lution as the standard solution. Perform the test with exactly
automatic integration method: the area of the peak, having 20 mL each of the sample solution and standard solution as
the relative retention time of about 1.9 to betahistine ob- directed under Liquid Chromatography <2.01> according to
tained from the sample solution, is not larger than 3/5 times the following conditions, and determine the peak areas, AT
the peak area of betahistine obtained from the standard so- and AS, of betahistine in each solution.
lution, and the total area of the peaks other than betahistine
of betahistine mesilate (C8H12N2.2CH4O3S)
betahistine from the standard solution.
＝ MS × AT/AS × V?/V × 1/C × 36
Detector, column, column temperature, mobile phase, and MS: Amount (mg) of betahistine mesilate for assay taken
flow rate: Proceed as directed in the operating conditions in C: Labeled amount (mg) of betahistine mesilate
the Assay. (C8H12N2.2CH4O3S) in 1 tablet
retention time of betahistine, beginning after the solvent
peak.
Assay.
standard solution add the mixture of water and acetonitrile
(63:37) to make exactly 50 mL. Confirm that the peak area
of betahistine obtained with 20 mL of this solution is equiva-
factor of the peak of betahistine are not less than 2000 and
lent to 7 to 13z of that obtained with 20 mL of the standard
solution.
System performance: Dissolve 10 mg of betahistine mesi-
JP XVII Official Monographs / Betamethasone 497

ing conditions, the relative standard deviation of the peak Betamethasone
area of betahistine is not more than 2.0z.
ベタメタゾン
Assay Weigh accurately the mass of not less than 20 Beta-
histine Mesilate Tablets, and powder. Weigh accurately a
portion of the powder, equivalent to about 20 mg of beta-
histine mesilate (C8H12N2.2CH4O3S), add 40 mL of 0.1
mol/L hydrochloric acid TS, agitate for 10 minutes with the
aid of ultrasonic waves, and add 0.1 mol/L hydrochloric
acid TS to make exactly 50 mL. Centrifuge, and use the
supernatant liquid as the sample solution. Separately, weigh
C22H29FO5: 392.46
accurately about 0.1 g of betahistine mesilate for assay, pre-
9-Fluoro-11b,17,21-trihydroxy-16b-methylpregna-
viously dried under reduced pressure with phosphorous (V)
1,4-diene-3,20-dione
oxide at 709C for 24 hours, and dissolve in 0.1 mol/L hydro-
[378-44-9]
chloric acid TS to make exactly 50 mL. Pipet 10 mL of this
solution, add 0.1 mol/L hydrochloric acid TS to make ex-
Betamethasone, when dried, contains not less than
96.0z and not more than 103.0z of betamethasone
(C22H29FO5).
tography <2.01>, according to the following conditions, and Description Betamethasone occurs as a white to pale yel-
determine the peak areas, AT and AS, of betahistine in each lowish white crystalline powder.
solution. It is sparingly soluble in methanol, in ethanol (95) and in
acetone, and practically insoluble in water.
Amount (mg) of betahistine mesilate (C8H12N2.2CH4O3S)
＝ MS × AT/AS × 1/5
MS: Amount (mg) of betahistine mesilate for assay taken
Identification (1) Proceed with 10 mg of Betamethasone
Operating conditions— as directed under Oxygen Flask Combustion Method <1.06>,
Detector: An ultraviolet absorption photometer (wave- using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide
length: 261 nm). TS and 20 mL of water as the absorbing liquid: the test solu-
Column: A stainless steel column 4.6 mm in inside diame- tion so obtained responds to the Qualitative Tests <1.09> for
ter and 15 cm in length, packed with octadecylsilanized silica fluoride.
gel for liquid chromatography (5 mm in particle diameter). (2) Dissolve 1.0 mg of Betamethasone in 10 mL of
Column temperature: A constant temperature of about ethanol (95). Mix 2.0 mL of the solution with 10 mL of
359 C. phenylhydrazinium hydrochloride TS, heat in a water bath
Mobile phase: To 5 mL of diethylamine and 20 mL of at 609C for 20 minutes, and cool the solution. Determine the
acetic acid (100) add water to make 1000 mL. In 630 mL of absorption spectrum of the solution as directed under Ultra-
this solution dissolve 2.3 g of sodium lauryl sulfate, and add violet-visible Spectrophotometry <2.24>, using as the blank
370 mL of acetonitrile. the solution prepared with 2.0 mL of ethanol (95) in the
Flow rate: Adjust so that the retention time of betahistine same manner as the former solution, and compare the spec-
is about 5 minutes. trum with the Reference Spectrum or the spectrum of a solu-
System suitability— tion of Betamethasone RS prepared in the same manner as
System performance: When the procedure is run with 5 mL the sample solution: both spectra exhibit similar intensities
of the standard solution under the above operating condi- of absorption at the same wavelengths.
tions, the number of theoretical plates and the symmetry fac- (3) Determine the infrared absorption spectrum of Beta-
tor of the peak of betahistine are not less than 2000 and not methasone, previously dried, as directed in the potassium
more than 1.5, respectively. bromide disk method under Infrared Spectrophotometry
System repeatability: When the test is repeated 6 times <2.25>, and compare the spectrum with the Reference Spec-
with 5 mL of the standard solution under the above operating trum or the spectrum of previously dried Betamethasone RS:
conditions, the relative standard deviation of the peak area both spectra exhibit similar intensities of absorption at the
of betahistine is not more than 1.0z. same wave numbers. If any difference appears between the
spectra, dissolve Betamethasone and Betamethasone RS in
acetone, respectively, then evaporate the acetone to dryness,
and repeat the test on the residues.
0.1 g, methanol, 20 mL, 100 mm).
Betamethasone according to Method 2, and perform the test.
(2) Related substances—Dissolve 10 mg of Betametha-
sone in 5 mL of a mixture of chloroform and methanol (9:1),
the sample solution, add a mixture of chloroform and meth-
anol (9:1) to make exactly 100 mL, and use this solution as
498 Betamethasone Tablets / Official Monographs JP XVII
as directed under Thin-layer Chromatography <2.03>. Spot 5
mL each of the sample solution and standard solution on a Betamethasone Tablets
chromatography. Develop the plate with a mixture of ベタメタゾン錠
dichloromethane, diethyl ether, methanol and water
(385:75:40:6) to a distance of about 12 cm, and air-dry the
Betamethasone Tablets contain not less than 90.0z
plate. Examine under ultraviolet light (main wavelength: 254
nm): the spots other than the principal spot from the sample
betamethasone (C22H29FO5: 392.46).
ard solution. Method of preparation Prepare as directed under Tablets,
with Betamethasone.
um, phosphorus (V) oxide, 4 hours). Identification Pulverize Betamethasone Tablets. To a por-
tion of the powder, equivalent to 2 mg of Betamethasone,
Residue on ignition <2.44> Not more than 0.5z (0.1 g,
add 20 mL of methanol, shake for 5 minutes, and filter.
platinum crucible).
Evaporate the filtrate on a water bath to dryness, dissolve
Assay Dissolve about 20 mg each of Betamethasone and the residue after cooling in 2 mL of methanol, filter if neces-
Betamethasone RS, previously dried and accurately weighed, sary, and use this as the sample solution. Separately, dissolve
in methanol to make exactly 50 mL. Pipet 5 mL each of 2 mg of Betamethasone RS in 2 mL of methanol, and use
these solutions, add exactly 5 mL each of the internal stand- this solution as the standard solution. Perform the test with
ard solution, then add methanol to make 50 mL, and use these solutions as directed under Thin-layer Chromatogra-
these solutions as the sample solution and standard solution, phy <2.03>. Spot 5 mL each of the sample solution and stand-
respectively. Perform the test with 10 mL each of these solu- ard solution on a plate of silica gel with fluorescent indicator
tions as directed under Liquid Chromatography <2.01> ac- for thin-layer chromatography, develop with a mixture of 1-
cording to the following conditions, and calculate the ratios, butanol, water and acetic anhydride (3:1:1) to a distance of
QT and QS, of the peak area of betamethasone to that of the about 10 cm, and air-dry the plate. Examine under ultravio-
internal standard. let light (main wavelength: 254 nm): the principal spot ob-
tained with the sample solution and the spot obtained with
Amount (mg) of betamethasone (C22H29FO5)
the standard solution show the same R f value.
＝ M S × Q T / QS
MS: Amount (mg) of Betamethasone RS taken
Internal standard solution—A solution of butyl parahy- Content uniformity test.
droxybenzoate in methanol (1 in 1750). To 1 tablet of Betamethasone Tablets add V mL of water
Operating conditions— so that each mL contains about 50 mg of betamethasone
Detector: An ultraviolet absorption photometer (wave- (C22H29FO5). Add exactly 2V mL of the internal standard so-
length: 240 nm). lution, shake vigorously for 10 minutes, centrifuge, and use
Column: A stainless steel column about 4.0 mm in inside the supernatant liquid as the sample solution. Separately,
diameter and 15 cm in length, packed with octadecylsilanized weigh accurately about 20 mg of Betamethasone RS, previ-
silica gel for liquid chromatography (5 mm in particle diame- ously dried for 4 hours in a desiccator (in vacuum, phospho-
ter). rus (V) oxide), dissolve in acetonitrile to make exactly 200
Column temperature: A constant temperature of about mL. Pipet 5 mL of this solution, add exactly 20 mL of the
259 C. internal standard solution, add 5 mL of water, and use this
Mobile phase: A mixture of water and acetonitrile (3:2). solution as the standard solution. Perform the test with 20
Flow rate: Adjust so that the retention time of betametha- mL each of the sample solution and standard solution as
sone is about 4 minutes. directed under Liquid Chromatography <2.01> according to
System suitability— the following conditions, and calculate the ratios, QT and
System performance: When proceed the test with 10 mL of QS, of the peak area of betamethasone to that of the internal
the standard solution under the above operating conditions, standard.
betamethasone and the internal standard are eluted in this
Amount (mg) of betamethasone (C22H29FO5)
＝ MS × QT/QS × V/400
than 10.
System repeatability: When the test is repeated 6 times MS: Amount (mg) of Betamethasone RS taken
Internal standard solution—A solution of butyl parahy-
droxybenzoate in acetonitrile (1 in 40,000).
the peak area of betamethasone to that of the internal stand-
ard is not more than 1.0z.
Containers and storage Containers—Tight containers. Assay.
Storage—Light-resistant. System suitability—
ditions, betamethasone and the internal standard are eluted
not less than 10.
JP XVII Official Monographs / Betamethasone Dipropionate 499
the peak area of betamethasone to that of the internal stand- actly 50 mL. Pipet 5 mL of this solution, add exactly 20 mL
ard is not more than 1.0z. of the internal standard solution and 5 mL of water, and use
in 30 minutes of Betamethasone Tablets is not less than
QS, of the peak area of betamethasone to that of the internal
85z.
standard.
Start the test with 1 tablet of Betamethasone Tablets,
withdraw not less than 20 mL of the medium at the specified Amount (mg) of betamethasone (C22H29FO5)
minute after starting the test, and filter through a membrane ＝ MS × QT/QS × 1/4
MS: Amount (mg) of Betamethasone RS taken
first 10 mL of the filtrate, pipet the subsequent V mL of the
filtrate, add water to make exactly V? mL so that each mL Internal standard solution—A solution of butyl parahy-
contains about 0.56 mg of betamethasone (C22H29FO5), and droxybenzoate in acetonitrile (1 in 10,000).
use this solution as the sample solution. Separately, weigh Operating conditions—
accurately about 28 mg of Betamethasone RS, previously Detector: An ultraviolet absorption photometer (wave-
dried in a desiccator (in vacuum, phosphorus (V) oxide) for 4 length: 240 nm).
hours, dissolve in methanol to make exactly 100 mL. Pipet 5 Column: A stainless steel column 4 mm in inside diameter
mL of this solution, and add water to make exactly 100 mL. and 15 cm in length, packed with octadecylsilanized silica gel
Pipet 4 mL of this solution, add water to make exactly 100 for liquid chromatography (5 mm in particle diameter).
mL, and use this solution as the standard solution. Perform Column temperature: A constant temperature of about
the test with exactly 100 mL each of the sample solution and 259C.
standard solution as directed under Liquid Chromatography Mobile phase: A mixture of water and acetonitrile (3:2).
<2.01> according to the following conditions, and determine Flow rate: Adjust so that the retention time of betametha-
the peak areas, AT and AS, of betamethasone in each solu- sone is about 4 minutes.
tion. System suitability—
of betamethasone (C22H29FO5)
ditions, betamethasone and the internal standard are eluted
＝ MS × AT/AS × V?/V × 1/C × 9/5
MS: Amount (mg) of Betamethasone RS taken not less than 10.
C: Labeled amount (mg) of betamethasone (C22H29FO5) in System repeatability: When the test is repeated 6 times
1 tablet with 20 mL of the standard solution under the above operat-
the peak area of betamethasone to that of the internal stand-
length: 241 nm).
Column: A stainless steel column 4.6 mm in inside diame- Containers and storage Containers—Tight containers.
ter and 15 cm in length, packed with octadecylsilanized silica Storage—Light-resistant.
259 C. Betamethasone Dipropionate
Mobile phase: A mixture of methanol and water (3:2).
Flow rate: Adjust so that the retention time of betametha- ベタメタゾンジプロピオン酸エステル
sone is about 7 minutes.
factor of the peak of betamethasone are not less than 3000
C28H37FO7: 504.59
area of betamethasone is not more than 2.0z.
9-Fluoro-11b,17,21-trihydroxy-16b-methylpregna-1,4-
Assay Weigh accurately the mass of not less than 20 Beta- diene-3,20-dione 17,21-dipropanoate
methasone Tablets, and powder. Weigh accurately a portion [5593-20-4]
of the powder, equivalent to about 5 mg of betamethasone
(C22H29FO5), add 25 mL of water, then add exactly 50 mL of Betamethasone Dipropionate, when dried, contains
the internal standard solution, and shake vigorously for 10 not less than 97.0z and not more than 103.0z of
minutes. Filter through a membrane filter with pore size not betamethasone dipropionate (C28H37FO7), and not less
exceeding 0.5 mm, discard the first 5 mL of the filtrate, and than 3.4z and not more than 4.1z of fluorine
use the subsequent filtrate as the sample solution. Sepa- (F:19.00).
rately, weigh accurately about 20 mg of Betamethasone RS,
Description Betamethasone Dipropionate occurs as a white
previously dried in a desiccator (in vacuum, phosphorus (V)
to pale yellowish white crystalline powder. It is odorless.
oxide) for 4 hours, and dissolve in acetonitrile to make ex-
It is freely soluble in acetone, in 1,4-dioxane and in chlo-
500 Betamethasone Sodium Phosphate / Official Monographs JP XVII
roform, soluble in methanol, sparingly soluble in ethanol and standard solution on a plate of silica gel with fluorescent
(95), slightly soluble in diethyl ether, and practically insolu- indicator for thin-layer chromatography. Develop the plate
ble in water and in hexane. with a mixture of chloroform and acetone (7:1) to a distance
It is affected gradually by light. of about 10 cm, and air-dry the plate. Examine under ultra-
violet light (main wavelength: 254 nm): the spots other than
Identification (1) To 1 mL of a solution of Betametha-
the principal spot from the sample solution are not more in-
sone Dipropionate in methanol (1 in 10,000) add 4 mL of
tense than the spot from the standard solution.
isoniazid TS, and heat on a water bath for 2 minutes: a
yellow color develops. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C,
(2) Proceed with 0.01 g of Betamethasone Dipropionate 3 hours).
as directed under Oxygen Flask Combustion Method <1.06>,
Residue on ignition <2.44> Not more than 0.2z (0.5 g,
using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide
platinum crucible).
TS and 20 mL of water as the absorbing liquid: the test solu-
tion so obtained responds to the Qualitative Tests <1.09> for Assay (1) Betamethasone dipropionate—Weigh accu-
fluoride. rately about 15 mg of Betamethasone Dipropionate, previ-
(3) Determine the absorption spectrum of a solution of ously dried, and dissolve in methanol to make exactly 100
Betamethasone Dipropionate in methanol (3 in 200,000) as mL. Pipet 5 mL of this solution, and dilute with methanol to
directed under Ultraviolet-visible Spectrophotometry <2.24>, exactly 50 mL. Determine the absorbance A of this solution
and compare the spectrum with the Reference Spectrum: at the wavelength of maximum absorption at about 239 nm
both spectra exhibit similar intensities of absorption at the as directed under Ultraviolet-visible Spectrophotometry
same wavelengths. <2.24>.
(4) Determine the infrared absorption spectrum of Beta-
Amount (mg) of betamethasone dipropionate (C28H37FO7)
methasone Dipropionate, previously dried, as directed in the
＝ A/312 × 10,000
potassium bromide disk method under Infrared Spectropho-
tometry <2.25>, and compare the spectrum with the Refer- (2) Fluorine—Weigh accurately about 10 mg of Beta-
ence Spectrum: both spectra exhibit similar intensities of ab- methasone Dipropionate, previously dried, and proceed as
sorption at the same wave numbers. directed in the procedure of determination for fluorine under
Oxygen Flask Combustion Method <1.06>, using a mixture
D : ＋63 – ＋709 (after drying,
of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of
50 mg, 1,4-dioxane, 10 mL, 100 mm).
water as the absorbing liquid.
Purity (1) Fluoride—To 0.10 g of Betamethasone Dipro- Storage—Light-resistant.
pionate add 10.0 mL of diluted 0.01 mol/L sodium hydrox-
ide TS (1 in 20), shake for 10 minutes, and filter through a
membrane filter (0.4-mm pore size). Place 5.0 mL of the fil- Betamethasone Sodium Phosphate
trate in a 20-mL volumetric flask, and add 10 mL of a mix-
ture of alizalin complexone TS, acetic acid-potassium acetate ベタメタゾンリン酸エステルナトリウム
buffer solution (pH 4.3) and cerium (III) nitrate TS (1:1:1),
add water to make 20 mL, allow to stand for 1 hour, and use
this solution as the sample solution. Separately, place 1.0
mL of Standard Fluorine Solution in a 20-mL volumetric
flask, add 5.0 mL of diluted 0.01 mol/L sodium hydroxide
TS (1 in 20), then 10 mL of a mixture of alizalin complexone
TS, acetic acid-potassium acetate buffer solution (pH 4.3)
and cerium (III) nitrate TS (1:1:1), proceed in the same man- C22H28FNa2O8P: 516.40
ner as the preparation of the sample solution, and use this 9-Fluoro-11b,17,21-trihydroxy-16b-methylpregna-
solution as the standard solution. Place 5.0 mL of diluted 1,4-diene-3,20-dione 21-(disodium phosphate)
0.01 mol/L sodium hydroxide TS (1 in 20) in a 20-mL volu- [151-73-5]
metric flask, and proceed in the same manner as the prepara-
tion of the sample solution. Using this solution as the blank, Betamethasone Sodium Phosphate contains not less
determine the absorbances of the sample solution and stand- than 97.0z and not more than 103.0z of betametha-
ard solution at 600 nm as directed under Ultraviolet-visible sone sodium phosphate (C22H28FNa2O8P), calculated
Spectrophotometry <2.24>: the absorbance of the sample so- on the anhydrous basis.
lution is not more than that of the standard solution (not
Description Betamethasone Sodium Phosphate occurs as
more than 0.012z).
white to pale yellowish white, crystalline powder or masses.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Beta-
It is odorless.
methasone Dipropionate according to Method 2, and per-
It is freely soluble in water, sparingly soluble in methanol,
slightly soluble in ethanol (95), and practically insoluble in
diethyl ether.
It is hygroscopic.
exposure to light, using light-resistant vessels. Dissolve 10
mg of Betamethasone Dipropionate in 10 mL of chloroform,
and use this solution as the sample solution. Pipet 3 mL of Identification (1) Dissolve 2 mg of Betamethasone So-
the sample solution, add chloroform to make exactly 100 dium Phosphate in 2 mL of sulfuric acid: a brown color de-
mL, and use this solution as the standard solution. Perform velops, and gradually changes to blackish brown.
the test as directed under Thin-layer Chromatography <2.03> (2) Prepare the test solution with 0.01 g of Betametha-
with these solutions. Spot 20 mL each of the sample solution sone Sodium Phosphate as directed under Oxygen Flask
JP XVII Official Monographs / Betamethasone Sodium Phosphate 501
Combustion Method <1.06>, using a mixture of 0.5 mL of wavelength: 254 nm): the spot from the sample solution
0.01 mol/L sodium hydroxide TS and 20 mL of water as an corresponding to the spot from the standard solution is not
absorbing liquid: the test solution responds to the Qualita- more intense than the spot from the standard solution.
tive Tests <1.09> (2) for fluoride.
Water <2.48> Not more than 10.0z (0.2 g, volumetric
(3) Take 40 mg of Betamethasone Sodium Phosphate in
titration, back titration).
a platinum crucible, and carbonize by heating. After cool-
ing, add 5 drops of nitric acid, and incinerate by heating. To Assay Weigh accurately about 20 mg each of Betametha-
the residue add 10 mL of diluted nitric acid (1 in 50), and sone Sodium Phosphate and Betamethasone Sodium Phos-
boil for several minutes. After cooling, filter if necessary, phate RS (separately, determine the water <2.48> in the same
and use this solution as the sample solution. The sample so- manner as Betamethasone Sodium Phosphate), and dissolve
lution responds to the Qualitative Tests <1.09> (2) for phos- each in methanol to make exactly 20 mL. Pipet 5 mL each of
phate. The sample solution neutralized with ammonia TS these solutions, and exactly 5 mL of the internal standard
responds to the Qualitative Tests <1.09> for sodium salt, and solution, then add methanol to make 50 mL, and use these
to the Qualitative Tests <1.09> (1) and (3) for phosphate. solutions as the sample solution and standard solution,
(4) Determine the infrared absorption spectrum of Beta- respectively. Perform the test with 10 mL each of the sample
methasone Sodium Phosphate, as directed in the potassium solution and standard solution as directed under Liquid
bromide disk method under Infrared Spectrophotometry Chromatography <2.01> according to the following condi-
<2.25>, and compare the spectrum with the Reference Spec- tions, and calculate the ratios, QT and QS, of the peak area
trum or the spectrum of Betamethasone Sodium Phosphate of betamethasone phosphate to that of the internal standard.
RS: both spectra exhibit similar intensities of absorption at
Amount (mg) of betamethasone sodium phosphate
(C22H28FNa2O8P) ＝ MS × QT/QS
D : ＋99 – ＋1059 (0.1 g calcu-
MS: Amount (mg) of Betamethasone Sodium Phosphate
RS taken, calculated on the anhydrous basis
pH <2.54> Dissolve 0.10 g of Betamethasone Sodium Phos-
phate in 20 mL of water: the pH of this solution is between
7.5 and 9.0.
Purity (1) Clarity and color of solution—Dissolve 0.25 g Detector: An ultraviolet absorption photometer (wave-
of Betamethasone Sodium Phosphate in 10 mL of water: the length: 254 nm).
solution is clear and colorless. Column: A stainless steel column 4.0 mm in inside diame-
(2) Free phosphoric acid—Weigh accurately about 20 mg ter and 25 cm in length, packed with octadecylsilanized silica
of Betamethasone Sodium Phosphate, dissolve in 20 mL of gel for liquid chromatography (7 mm in particle diameter).
water, and use this solution as the sample solution. Sepa- Column temperature: A constant temperature of about
rately, pipet 4 mL of Standard Phosphoric Acid Solution, 259C.
add 20 mL of water, and use this solution as the standard so- Mobile phase: Dissolve 1.6 g of tetra-n-butylammonium
lution. To each of the sample solution and the standard solu- bromide, 3.2 g of disodium hydrogen phosphate dodecahy-
tion add exactly 7 mL of dilute sulfuric acid, exactly 2 mL of drate and 6.9 g of potassium dihydrogen phosphate in 1000
hexaammonium heptamolybdate-sulfuric acid TS and ex- mL of water, and add 1500 mL of methanol.
actly 2 mL of p-methylaminophenol sulfate TS, shake well, Flow rate: Adjust so that the retention time of betametha-
and allow to stand at 20 ± 19C for 15 minutes. To each add sone phosphate is about 5 minutes.
water to make exactly 50 mL, and allow to stand at 20 ± System suitability—
19C for 15 minutes. Perform the test with these solutions as System performance: When the procedure is run with 10
directed under Ultraviolet-visible Spectrophotometry <2.24>, mL of the standard solution under the above operating con-
using a solution prepared with 20 mL of water in the same ditions, betamethasone phosphate and the internal standard
manner as the blank. Determine the absorbances, AT and are eluted in this order with the resolution between these
AS, of each solution from the sample solution and standard peaks being not less than 10.
solution at 730 nm: the amount of free phosphoric acid is System repeatability: When the test is repeated 6 times
not more than 0.5z. with 10 mL of the standard solution under the above operat-
Amount (z) of free phosphoric acid (H3PO4)
of the peak area of betamethasone phosphate to that of the
＝ AT/AS × 1/M × 10.32
internal standard is not more than 1.0z.
M: Amount (mg) of Betamethasone Sodium Phosphate
(3) Betamethasone—Dissolve 20 mg of Betamethasone
Sodium Phosphate in exactly 2 mL of methanol, and use this
of Betamethasone RS in exactly 10 mL of methanol. Pipet 1
mL of this solution, add methanol to make exactly 20 mL,
test with these solutions as directed under Thin-layer Chro-
and standard solution on a plate of silica gel with fluorescent
with a freshly prepared mixture of 1-butanol, water and
acetic anhydride (3:1:1) to a distance of about 10 cm, and
air-dry the plate. Examine under ultraviolet light (main
502 Betamethasone Valerate / Official Monographs JP XVII
accurately weighed, in methanol to make exactly 100 mL.
Betamethasone Valerate Pipet 10 mL each of these solutions, add 10 mL each of the
internal standard solution, and use these solutions as the
ベタメタゾン吉草酸エステル sample solution and the standard solution, respectively. Per-
form the test with 10 mL each of the sample solution and
the ratios, QT and QS, of the peak area of betamethasone
valerate to that of the internal standard.
Amount (mg) of betamethasone valerate (C27H37FO6)
＝ M S × Q T / QS
C27H37FO6: 476.58 MS: Amount (mg) of Betamethasone Valerate RS taken
9-Fluoro-11b,17,21-trihydroxy-16b-methylpregna-1,4-
Internal standard solution—A solution of isoamyl benzoate
diene-3,20-dione 17-pentanoate
in methanol (1 in 1000).
[2152-44-5]
Betamethasone Valerate, when dried, contains not
length: 254 nm).
betamethasone valerate (C27H37FO6).
Description Betamethasone Valerate occurs as a white crys- gel for liquid chromatography (7 mm in particle diameter).
talline powder. It is odorless. Column temperature: A constant temperature of about
It is freely soluble in chloroform, soluble in ethanol (95), 259C.
sparingly soluble in methanol, slightly soluble in diethyl Mobile phase: A mixture of methanol and water (7:3).
ether, and practically insoluble in water. Flow rate: Adjust so that the retention time of betametha-
Melting point: about 1909C (with decomposition). sone valerate is about 10 minutes.
Identification (1) Proceed with 0.01 g of Betamethasone
Valerate as directed under Oxygen Flask Combustion
Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L
ditions, betamethasone valerate and the internal standard
sodium hydroxide TS and 20 mL of water as the absorbing
liquid: the test solution so obtained responds to the Qualita-
tive Tests <1.09> for fluoride.
methasone Valerate, previously dried, as directed in the
potassium bromide disk method under Infrared Spectropho-
the peak area of betamethasone valerate to that of the inter-
nal standard is not more than 1.0z.
ence Spectrum or the spectrum of dried Betamethasone
Valerate RS: both spectra exhibit similar intensities of ab- Containers and storage Containers—Tight containers.
0.1 g, methanol, 20 mL, 100 mm). Betamethasone Valerate and
Purity Related substances—Conduct this procedure with- Gentamicin Sulfate Cream
out exposure to daylight. Dissolve 0.02 g of Betamethasone
ベタメタゾン吉草酸エステル･ゲンタマイシン硫酸塩ク
Valerate in 5 mL of a mixture of chloroform and methanol
リーム
mL of the sample solution, add a mixture of chloroform and
methanol (9:1) to make exactly 50 mL, and use this solution Betamethasone Valerate and Gentamicin Sulfate
as the standard solution. Perform the test with these solu- Cream contains not less than 90.0z and not more
tions as directed under Thin-layer Chromatography <2.03>. than 110.0z of the labeled amount of betamethasone
Spot 5 mL each of the sample solution and standard solution valerate (C27H37FO6: 476.58) and not less than 90.0z
on a plate of silica gel for thin-layer chromatography. De- and not more than 115.0z of the labeled amount of
velop the plate with a mixture of chloroform and methanol gentamicin C1(C21H43N5O7: 477.60).
(9:1) to a distance of about 12 cm, and air-dry the plate.
Method of preparation Prepare as directed under Creams,
Spray evenly alkaline blue tetrazolium TS on the plate: the
with Betamethasone Valerate and Gentamicin Sulfate.
spots other than the principal spot from the sample solution
are not more intense than the spot from the standard solu- Identification (1) To a quantity of Betamethasone Valer-
tion. ate and Gentamicin Sulfate Cream, equivalent to about 1.2
mg of Betamethasone Valerate, add 20 mL of methanol and
20 mL of hexane, shake vigorously for 10 minutes, and allow
3 hours).
to stand. Take 15 mL of the lower layer, evaporate the layer
Residue on ignition <2.44> Not more than 0.2z (0.5 g, to dryness on a water bath under a current of nitrogen. To
platinum crucible). the residue add 1 mL of ethyl acetate, mix, and use as the
sample solution. Separately, dissolve about 18 mg of Be-
Assay Dissolve about 10 mg each of Betamethasone Valer-
tamethasone Valerate RS in 20 mL of ethyl acetate, and use
ate and Betamethasone Valerate RS, previously dried and
JP XVII Official Monographs / Betamethasone Valerate and Gentamicin Sulfate Cream 503
these solutions as directed under Thin-layer Chromatogra- suitability test.

phy <2.03>. Spot 5 mL each of the sample solution and stand- System performance: When the procedure is run with 150
ard solution on a plate of silica gel for thin-layer chromatog- mL of the solution for system suitability test under the above
raphy, develop the plate with ethyl acetate to a distance of operating conditions, the number of theoretical plates and
about 10 cm, and air-dry the plate. Spray evenly alkaline the symmetry factor of the peak of betamethasone valerate
blue tetrazolium TS on the plate, and heat at 1009C: the are not less than 4000 and 0.8 to 1.3, respectively.
principal spot with the sample solution and the spot with the System repeatability: When the test is repeated 6 times
standard solution are purple in color, and their R f values are with 150 mL of the solution for system suitability test under
the same. the above operating conditions, the relative standard devia-
(2) To a quantity of Betamethasone Valerate and Gen- tion of the peak area of betamethasone valerate is not more
tamicin Sulfate Cream, equivalent to about 2 mg (potency) than 2.0z.
of Gentamicin Sulfate, add 20 mL of ethyl acetate and 10
Assay (1) Betamethasone valerate—Weigh accurately an
mL of water, shake vigorously for 10 minutes, and centri-
amount of Betamethasone Valerate and Gentamicin Sulfate
fuge. To 3 mL of the lower layer add 1 mL of dilute sodium
Cream, equivqlent to about 1 mg of betamethasone valerate
hydroxide TS and 2 mL of ninhydrin TS, and heat in a water
(C27H37FO6), add 10 mL of a mixture of methanol and water
bath at 90 – 959C for 10 minutes: a purple to dark purple
(7:3), and add exactly 10 mL of the internal standard solu-
color develops.
tion. After warming in a water bath at 609 C for 5 minutes,
pH <2.54> To a quantity of Betamethasone Valerate and shake vigorously for 20 minutes. Repeat this procedure
Gentamicin Sulfate Cream, equivalent to 6 mg of Beta- twice, cool with ice for 15 minutes, centrifuge for 5 minutes,
methasone Valerate, add 15 mL of water, and mix while then filter the supernatant liquid, discard the first 5 mL of
warming on a water bath to make a milky liquid: the pH of filtrate, and use the subsequent filtrate as the sample solu-
the cooled liquid is between 4.0 and 6.0. tion. Separately, weigh accurately about 25 mg of Beta-
methasone Valerate RS, previously dried at 1059 C for 3
Purity Related substances—Weigh accurately an amount
hours, and dissolve in methanol to make exactly 25 mL.
of Betamethasone Valerate and Gentamicin Sulfate Cream,
Pipet 5 mL of this solution, and add the mixture of metha-
equivalent to about 1 mg of Betamethasone Valerate, and
nol and water (7:3) to make exactly 50 mL. Pipet 10 mL of
add 10 mL of a mixture of methanol and water (7:3). Warm
this solution, add exactly 10 mL of the internal standard so-
in a water bath at 609C for 5 minutes, and shake vigorously
lution, mix, and use this solution as the standard solution.
for 20 minutes. Repeat this procedure 2 times. After cooling
for 15 minutes with ice, centrifuge for 5 minutes, take away
the bubbles from the upper surface, and filter the remaining
liquid. Discard first 2 mL of the filtrate, and use the subse-
the ratios, QT and QS, of the peak area of betamethasone
quent filtrate as the sample solution. Perform the test with
valerate to that of the internal standard.
150 mL of the sample solution as directed under Liquid
Chromatography <2.01> according to the following condi- Amount (mg) of betamethasone valerate (C27H37FO6)
tions, determine each peak area by the automatic integration ＝ MS × QT/QS × 1/25
method, and calculate these amounts by the area percentage
MS: Amount (mg) of Betamethasone Valerate RS taken
method: the amount of the substance other than betametha-
sone valerate is not more than 3.5z, and the total amount Internal standard solution—Dissolve 20 mg of beclometa-
of them is not more than 7.0z. sone dipropionate in 10 mL of methanol, and add the mix-
Operating conditions— ture of methanol and water (7:3) to make 200 mL.
Detector: An ultraviolet absorption photometer (wave- Operating conditions—
length: 240 nm). Detector: An ultraviolet absorption photometer (wave-
Column: A stainless steel column 4.6 mm in inside diame- length: 254 nm).
ter and 15 cm in length, packed with octadecylsilanized silica Column: A stainless steel column 2.1 mm in inside diame-
gel for liquid chromatography (5 mm in particle diameter). ter and 10 cm in length, packed with octadecylsilanized silica
Column temperature: A constant temperature of about gel for liquid chromatography (3.5 mm in particle diameter).
459 C. Column temperature: A constant temperature of about
Mobile phase: A mixture of water, acetonitrile and metha- 259C.
nol (12:7:1). Mobile phase: A mixture of methanol and water (13:7).
Flow rate: Adjust so that the retention time of betametha- Flow rate: Adjust so that the retention time of betametha-
sone valerate is about 16 minutes. sone valerate is about 16 minutes.
Time span of measurement: About 2.5 times as long as the System suitability—
retention time of betamethasone valerate beginning after the System performance: When the procedure is run with 3 mL
solvent peak. The peaks of the compounding ingredients are of the standard solution under the above operating condi-
not determined. tions, betamethasone valerate and the internal standard are
System suitability— eluted in this order with the resolution between these peaks
Test for required detectability: Dissolve 20 mg of Beta- being not less than 4.
methasone Valerate in 100 mL of a mixture of methanol and System repeatability: When the test is repeated 6 times
water (7:3). To exactly 1 mL of this solution add the mixture with 3 mL of the standard solution under the above operating
of methanol and water (7:3) to make exactly 100 mL, and conditions, the relative standard deviation of the ratio of the
use this solution as the solution for system suitability test. To peak area of betamethasone valerate to that of the internal
exactly 2.5 mL of the solution for system suitability test add standard is not more than 1.0z.
the mixture of methanol and water (7:3) to make exactly 50 (2) Gentamicin sulfate—Perform the test according to
mL. Confirm that the peak area of betamethasone valerate the Cylinder-plate method as directed under Microbial Assay
obtained with 150 mL of this solution is equivalent to 3.5 to for Antibiotics <4.02> according to the following conditions.
6.5z of that obtained with 150 mL of the solution for system (i) Test organism, agar media for base layer and seed
504 Betamethasone Valerate and Gentamicin Sulfate Ointment / Official Monographs JP XVII
layer, agar medium for transferring test organisms, and water bath to dissolve. After cooling, separate the water
standard solutions—Proceed as directed in the Assay layer: the pH of the layer is between 4.0 and 7.0.
under Gentamicin Sulfate.
Assay (1) Betamethasone valerate—Weigh accurately an
(ii) Sample solutions—Weigh accurately an amount of
amount of Betamethasone Valerate and Gentamicin Sulfate
Betamethasone Valerate and Gentamicin Sulfate Cream,
Ointment, equivalent to about 1 mg of betamethasone valer-
equivalent to about 1 mg (potency) of Gentamicin Sulfate,
ate (C27H37FO6), add 10 mL of a mixture of methanol and
add 100 mL of 0.1 mol/L phosphate buffer solution (pH
water (7:3), and add exactly 10 mL of the internal standard
8.0) previously warmed to about 859 C, and shake well to
solution. After warming in a water bath at 759C for 5
dissolve. After cooling, add 0.1 mol/L phosphate buffer
minutes, shake vigorously for 10 minutes. Repeat this proce-
solution (pH 8.0) to make exactly 250 mL to make the
dure once more, cool with ice for 15 minutes, filter, discard
high concentration sample solution, which contains 4 mg
the first 5 mL of filtrate, and use the subsequent filtrate as
(potency) per mL. Pipet a suitable amount of the high
concentration sample solution, add 0.1 mol/L phosphate
mg of Betamethasone Valerate RS, previously dried at
buffer solution (pH 8.0) so that each mL contains 1 mg
1059C for 3 hours, and dissolve in methanol to make exactly
(potency), and use this solution as the low concentration
25 mL. Pipet 5 mL of this solution, and add the mixture of
sample solution.
methanol and water (7:3) to make exactly 50 mL. Pipet 10
Containers and storage Containers—Tight containers. mL of this solution, add exactly 10 mL of the internal stand-
Storage—Light-resistant. ard solution, mix, and use this solution as the standard solu-
tion. Perform the test with 3 mL each of the sample solution
Betamethasone Valerate and raphy <2.01> according to the following conditions, and cal-
culate the ratios, QT and QS, of the peak area of betametha-
Gentamicin Sulfate Ointment sone valerate to that of the internal standard.
ベタメタゾン吉草酸エステル･ゲンタマイシン硫酸塩軟膏 Amount (mg) of betamethasone valerate (C27H37FO6)
＝ MS × QT/QS × 1/25
Betamethasone Valerate and Gentamicin Sulfate MS: Amount (mg) of Betamethasone Valerate RS taken
Ointment contains not less than 95.0z and not more
Internal standard solution—Dissolve 20 mg of beclometa-
than 110.0z of the labeled amount of betamethasone
sone dipropionate in 10 mL of methanol, and add the mix-
valerate (C27H37FO6: 476.58) and not less than 90.0z
ture of methanol and water (7:3) to make 200 mL.
and not more than 115.0z of the labeled potency of
gentamicin C1 (C21H43N5O7: 477.60).
Method of preparation Prepare as directed under Oint- length: 254 nm).
ment, with Betamethasone Valerate and Gentamicin Sulfate. Column: A stainless steel column 2.1 mm in inside diame-
Identification (1) To a quantity of Betamethasone Valer-
gel for liquid chromatography (3.5 mm in particle diameter).
ate and Gentamicin Sulfate Ointment, equivalent to 1.2 mg
of Betamethasone Valerate, add 20 mL of methanol and 20
259C.
mL of hexane, and disperse the ointment with the aid of
Mobile phase: A mixture of methanol and water (13:7).
ultrasonic. Shake vigorously for 5 minutes, centrifuge for 5
Flow rate: Adjust so that the retention time of betametha-
minutes, cool for 15 minutes with ice, and take 15 mL of the
sone valerate is about 16 minutes.
lower layer. Evaporate the layer to dryness on a water bath
under a current of nitrogen. To the residue add 1 mL of
ethyl acetate, apply ultrasonic waves, filter, if necessary, and
use the filtrate as the sample solution. Separately, dissolve 18
tions, betamethasone valerate and the internal standard are
mg of Betamethasone Valerate RS in 20 mL of ethyl acetate,
being not less than 4.
and standard solution on a plate of silica gel for thin-layer
chromatography, develop the plate with ethyl acetate to a
peak area of betamethasone valerate to that of the internal
distance of about 10 cm, and air-dry the plate. Spray evenly
standard is not more than 1.0z.
alkaline blue tetrazolium TS on the plate, and heat at 1009C:
(2) Gentamicin sulfate—Perform the test according to
the principal spot from the sample solution and the spot
the Cylinder-plate method as directed under Microbial Assay
from the standard solution are purple in color, and their R f
for Antibiotics <4.02> according to the following conditions.
values are the same.
(i) Test organism, agar media for base layer and seed
(2) To a quantity of Betamethasone Valerate and Gen-
layer, agar medium for transferring test organisms, and
tamicin Sulfate Ointment, equivalent to 2 mg (potency) of
standard solutions—Proceed as directed in the Assay under
Gentamicin Sulfate, add 20 mL of hexane and 10 mL of
Gentamicin Sulfate.
water, shake vigorously for 10 minutes, and centrifuge. To
(ii) Sample solutions—Weigh accurately an amount of
3 mL of the lower layer add 1 mL of dilute sodium hydrox-
Betamethasone Valerate and Gentamicin Sulfate Ointment,
ide TS and 2 mL of ninhydrin TS, and heat in a water bath
equivalent to about 1 mg (potency) of Gentamicin Sulfate,
at 90 – 959C for 10 minutes: a red-brown color develops.
transfer to a separator, add 50 mL of petroleum ether and
pH <2.54> To a quantity of Betamethasone Valerate and exactly 100 mL of 0.1 mol/L phosphate buffer solution (pH
Gentamicin Sulfate Ointment, equivalent to 6 mg of Beta- 8.0), shake for 10 minutes, and allow to stand. Pipet a suita-
methasone Valerate, add 15 mL of water, and warm on a ble amount of the water layer, add 0.1 mol/L phosphate
JP XVII Official Monographs / Betamipron 505
buffer solution (pH 8.0) to make solutions so that each mL from the standard solution is not more intense than the spot
contains 4 mg (potency) and 1 mg (potency), and use these from the standard solution.
solutions as the high concentration sample solution and low (4) Related substances—Dissolve 20 mg of Betamipron
concentration sample solution, respectively. in 100 mL of the mobile phase, and use this solution as the
Betamipron lowing conditions, and determine each peak area by the au-
tomatic integration method: the area of the peak other than
ベタミプロン
betamipron from the sample solution is not larger than 2/5
times the peak area of betamipron from the standard solu-
tion, and the total area of the peaks other than betamipron
betamipron from the standard solution.
C10H11NO3: 193.20
3-Benzoylaminopropanoic acid
length: 225 nm).
[3440-28-6]
Betamipron contains not less than 99.0z and not
more than 101.0z of betamipron (C10H11NO3), calcu-
lated on the anhydrous basis.
409C.
Description Betamipron occurs as white, crystals or crys- Mobile phase: Dissolve 3.12 g of sodium dihydrogen phos-
talline powder. phate dihydrate in 800 mL of water, adjust to pH 7.0 with
It is freely soluble in methanol, soluble in ethanol (99.5), dilute sodium hydroxide TS, and add water to make 1000
and slightly soluble in water. mL. To 900 mL of this solution add 100 mL of acetonitrile.
It dissolves in sodium hydroxide TS. Flow rate: Adjust so that the retention time of betamipron
is about 6 minutes.
solution of Betamipron in ethanol (99.5) (1 in 100,000) as
retention time of betamipron, beginning after the solvent
peak.
same wavelengths.
mL. Confirm that the peak area of betamipron obtained
mipron as directed in the potassium bromide disk method
with 10 mL of this solution is equivalent to 7 to 13z of that
obtained with 10 mL of the standard solution.
System performance: Dissolve 5 mg of Betamipron and
5 mg of benzoic acid in 200 mL of the mobile phase. When
pH <2.54> Dissolve 0.25 g of Betamipron in 100 mL of the procedure is run with 10 mL of this solution under the
water by warming, and cool: the pH of this solution is be- above operating conditions, benzoic acid and betamipron
tween 3.0 and 3.4. are eluted in this order with the resolution between these
Purity (1) Clarity and color of solution—Dissolve 1.0 g with 10 mL of the standard solution under the above operat-
of Betamipron in 10 mL of sodium hydroxide TS: the solu- ing conditions, the relative standard deviation of the peak
tion is clear and colorless. area of betamipron is not more than 2.0z.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Beta-
mipron according to Method 4, and perform the test. Pre-
lution (not more than 10 ppm). Residue on ignition <2.44> Not more than 0.1z (1 g).
(3) b-Alanine—Dissolve 0.25 g of Betamipron in 10 mL
Assay Weigh accurately about 0.25 g of Betamipron, dis-
solve in 25 mL of ethanol (99.5), add 25 mL of water, and
Separately, dissolve 50 mg of b-alanine in methanol to make
titrate <2.50> with 0.1 mol/L sodium hydroxide VS (poten-
exactly 100 mL. Pipet 1 mL of this solution, add methanol
tiometric titration). Perform the blank determination in the
to make exactly 10 mL, and use this solution as the standard
under Thin-layer Chromatography <2.03>. Spot 5 mL each of Each mL of 0.1 mol/L sodium hydroxide VS
the sample solution and standard solution on a plate of silica ＝ 19.32 mg of C10H11NO3
gel for thin-layer chromatography, develop the plate with a
mixture of methanol, ethyl acetate, ammonia solution (28)
and water (200:200:63:37) to a distance of about 10 cm, and
air-dry the plate. Spray evenly ninhydrin-butanol TS on the
plate, and heat at 1059 C for 5 minutes: the spot obtained
from the sample solution corresponding to the spot obtained
506 Betaxolol Hydrochloride / Official Monographs JP XVII
than 3, and they are not more intense than the spot obtained
Betaxolol Hydrochloride from the standard solution.
(5) Related substance II—Dissolve 0.10 g of Betaxolol
ベタキソロール塩酸塩 Hydrochloride in 50 mL of the mobile phase, and use this
solution as the sample solution. Pipet 1 mL of the sample
C18H29NO3.HCl: 343.89 area of both solutions by the automatic integration method:
(2RS)-1-{4-[2-(Cyclopropylmethoxy)ethyl]phenoxy}- the area of the peak other than betaxolol obtained from the
3-[(1-methylethyl)amino]propan-2-ol monohydrochloride sample solution is not larger than the peak area of betaxolol
[63659-19-8] from the standard solution, and the total area of the peaks
other than the peak of betaxolol from the sample solution is
Betaxolol Hydrochloride, when dried, contains not not larger than 2 times the peak area of betaxolol from the
less than 99.0z and not more than 101.0z of standard solution.
betaxolol hydrochloride (C18H29NO3.HCl). Operating conditions—
Description Betaxolol Hydrochloride occurs as white, crys-
length: 273 nm).
It is very soluble in water, and freely soluble in methanol,
in ethanol (99.5) and in acetic acid (100).
Dissolve 1.0 g of Betaxolol Hydrochloride in 50 mL of
water: the pH of the solution is between 4.5 and 6.5.
259C.
A solution of Betaxolol Hydrochloride (1 in 100) shows no
Mobile phase: A mixture of diluted 0.05 mol/L potassium
optical rotation.
dihydrogen phosphate TS (1 in 2) with the pH adjusted to
Identification (1) Determine the absorption spectrum of a 3.0 with 1 mol/L hydrochloric acid TS, acetonitrile and
solution of Betaxolol Hydrochloride in ethanol (99.5) (1 in methanol (26:7:7).
10,000) as directed under Ultraviolet-visible Spectropho- Flow rate: Adjust so that the retention time of betaxolol is
tometry <2.24>, and compare the spectrum with the Refer- about 9 minutes.
ence Spectrum: both spectra exhibit similar intensities of ab- Time span of measurement: About 2 times as long as the
sorption at the same wavelengths. retention time of betaxolol, beginning after the solvent peak.
(2) Determine the infrared absorption spectrum of Be- System suitability—
taxolol Hydrochloride as directed in the potassium chloride Test for required detectability: Pipet 4 mL of the standard
disk method under Infrared Spectrophotometry <2.25>, and solution, and add the mobile phase to make exactly 20 mL.
compare the spectrum with the Reference Spectrum: both Confirm that the peak area of betaxolol obtained from 10
spectra exhibit similar intensities of absorption at the same mL of this solution is equivalent to 14 to 26z of that ob-
wave numbers. tained from 10 mL of the standard solution.
(3) A solution of Betaxolol Hydrochloride (1 in 10) re- System performance: Dissolve 50 mg of Betaxolol Hydro-
sponds to the Qualitative Tests <1.09> (2) for chloride. chloride and 5 mg of 2-naphthol in 200 mL of the mobile
phase. When the procedure is run with 10 mL of this solution
under the above operating conditions, betaxolol and 2-
Purity (1) Clarity and color of solution—Dissolve 1.0 g naphthol are eluted in this order with the resolution between
of Betaxolol Hydrochloride in 10 mL of water: the solution these peaks being not less than 10.
is clear and colorless. System repeatability: When the test is repeated 6 times
(2) Heavy metals <1.07>—Proceed with 2.0 g of Betax- with 10 mL of the standard solution under the above operat-
olol Hydrochloride according to Method 4, and perform the ing conditions, the relative standard deviation of the peak
test. Prepare the control solution with 2.0 mL of Standard area of betaxolol is not more than 2.0z.
4 hours).
of Betaxolol Hydrochloride according to Method 3, and per-
form the test (not more than 1 ppm). Residue on ignition <2.44> Not more than 0.1z (1 g).
(4) Related substance I—Dissolve 0.10 g of Betaxolol
Assay Weigh accurately about 0.3 g of Betaxolol Hydro-
chloride, previously dried, dissolve in 30 mL of acetic acid
(100), add 30 mL of acetic anhydride, and titrate <2.50> with
and add methanol to make exactly 50 mL. Pipet 1 mL of this
0.1 mol/L perchloric acid VS (potentiometric titration). Per-
<2.03>. Spot 10 mL each of the sample solution and standard Each mL of 0.1 mol/L perchloric acid VS
solution on a plate of silica gel for thin-layer chromatogra- ＝ 34.39 mg of C18H29NO3.HCl
phy. Develop the plate with a mixture of ethyl acetate, water
and acetic acid (100) (10:3:3) to a distance of about 10 cm,
and air-dry the plate. Allow the plate to stand in iodine
vapor for 1 hour: the number of the spots other than the
principal spot obtained from the sample solution is not more
JP XVII Official Monographs / Bezafibrate 507
Assay Weigh accurately about 0.4 g of Bethanechol Chlo-

Bethanechol Chloride ride, previously dried, dissolve in 2 mL of acetic acid (100),
ベタネコール塩化物 mol/L perchloric acid VS (potentiometric titration). Per-
tion.
＝ 19.67 mg of C7H17ClN2O2
C7H17ClN2O2: 196.68
(2RS )-2-Carbamoyloxy-N, N, N-
trimethylpropylaminium chloride
[590-63-6] Bezafibrate
ベザフィブラート
Bethanechol Chloride, when dried, contains not less
than 98.0z and not more than 101.0z of bethanechol
chloride (C7H17ClN2O2).
Description Bethanechol Chloride occurs as colorless or
white crystals or a white, crystalline powder.
It is very soluble in water, freely soluble in acetic acid
(100), and sparingly soluble in ethanol (99.5). C19H20ClNO4: 361.82
It is hygroscopic. 2-(4-{2-[(4-Chlorobenzoyl)amino]ethyl}phenoxy)-2-
A solution of Bethanechol Chloride (1 in 10) shows no methylpropanoic acid
optical rotation. [41859-67-0]
Identification (1) To 2 mL of a solution of Bethanechol
Bezafibrate, when dried, contains not less than
Chloride (1 in 40) add 0.1 mL of a solution of cobalt (II)
98.5z and not more than 101.0z of bezafibrate
chloride hexahydrate (1 in 100), then add 0.1 mL of potas-
(C19H20ClNO4).
sium hexacyanoferrate (II) TS: A green color is produced,
and almost entirely fades within 10 minutes. Description Bezafibrate occurs as a white crystalline pow-
(2) To 1 mL of a solution of Bethanechol Chloride (1 in der.
100) add 0.1 mL of iodine TS: a brown precipitate is pro- It is freely soluble in N, N-dimethylformamide, soluble in
duced, and the solution shows a greenish brown color. methanol, slightly soluble in ethanol (99.5), and practically
(3) Determine the infrared absorption spectrum of insoluble in water.
Bethanechol Chloride as directed in the paste method under
Infrared Spectrophotometry <2.25>, and compare the spec-
solution of Bezafibrate in methanol (1 in 100,000) as directed
trum with the Reference Spectrum: both spectra exhibit simi-
lar intensities of absorption at the same wave numbers.
(4) A solution of Bethanechol Chloride (1 in 100) re-
wavelengths.
Melting point <2.60> 217 – 2219C (after drying). (2) Determine the infrared absorption spectrum of
Bezafibrate, previously dried, as directed in the potassium
Bethanechol Chloride according to Method 1, and perform
(2) Related substances—Dissolve 1.0 g of Bethanechol
(3) Perform the test with Bezafibrate as directed under
Chloride in 2.5 mL of water, and use this solution as the
water to make exactly 100 mL, and use this solution as the Melting point <2.60> 181 – 1869C
Purity (1) Chloride <1.03>—Dissolve 3.0 g of Bezafibrate
in 15 mL of N, N-dimethylformamide, add water to make 60
mL, shake well, allow to stand for more than 12 hours, and
plate of cellulose for thin-layer chromatography. Develop
filter. To 40 mL of the filtrate add 6 mL of dilute nitric acid
the plate with a mixture of a solution of ammonium acetate
and water to make 50 mL, and perform the test using this
(1 in 100), acetone, 1-butanol and formic acid (20:20:20:1) to
a distance of about 10 cm, and dry the plate at 1059C for 15
follows: To 0.70 mL of 0.01 mol/L hydrochloric acid VS
minutes. Spray evenly hydrogen hexachloroplatinate (IV)-
add 10 mL of N, N-dimethylformamide, 6 mL of dilute nitric
potassium iodide TS on the plate, and allow to stand for 30
acid and water to make 50 mL (not more than 0.012z).
minutes: the spot other than the principal spot from the sam-
ple solution is not more intense than the spot from the stand-
Bezafibrate according to Method 4, and perform the test.
ard solution.
Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, Solution (not more than 10 ppm).
2 hours). (3) Related substances—Dissolve 0.10 g of Bezafibrate in
35 mL of methanol, add diluted 0.5 mol/L ammonium ace-
tate TS (1 in 50) to make 50 mL, and use this solution as the
508 Bezafibrate Extended-release Tablets / Official Monographs JP XVII
sample solution. Pipet 1 mL of the sample solution, add 70
mL of methanol and diluted 0.5 mol/L ammonium acetate Bezafibrate Extended-release
TS (1 in 50) to make exactly 100 mL, and use this solution as
the standard solution. Perform the test with exactly 5 mL Tablets
ベザフィブラート徐放錠
lowing conditions, and determine each peak area by the au-
tomatic integration method: the areas of the peaks having Bezafibrate Extended-release Tablets contain not
the relative retention times of about 0.65 and 1.86 to less than 95.0z and not more than 105.0z of the
bezafibrate obtained from the sample solution are not larger labeled amount of bezafibrate (C19H20ClNO4: 361.82).
than 1/2 times the peak area of bezafibrate obtained from
the standard solution, the area of the peak other than those
with Bezafibrate.
and other than bezafibrate from the sample solution is not
larger than 1/5 times the peak area of bezafibrate from the Identification Mix well an amount of powdered
standard solution, and the total area of the peaks other than Bezafibrate Extended-release Tablets, equivalent to 0.1 g of
the peak of bezafibrate from the sample solution is not Bezafibrate, with 100 mL of methanol, and filter. To 1 mL
larger than 3/4 times the peak area of bezafibrate from the of the filtrate add methanol to make 100 mL. Determine the
standard solution. absorption spectrum of this solution as directed under Ultra-
Operating conditions— violet-visible Spectrophotometry <2.24>: it exhibits a maxi-
Detector: An ultraviolet absorption photometer (wave- mum between 227 nm and 231 nm.
length: 230 nm).
gel for liquid chromatography (5 mm in particle diameter). Dissolution <6.10> When the test is performed at 50 revolu-
Column temperature: A constant temperature of about tions per minute according to the Paddle method, using 900
259 C. mL of disodium hydrogen phosphate-citric acid buffer solu-
Mobile phase: A mixture of methanol and diluted acetic tion (pH 7.2) as the dissolution medium, the dissolution
acid (100) (1 in 100) (9:4). rates of a 100-mg tablet in 1.5 hours, in 2.5 hours and in 8
Flow rate: Adjust so that the retention time of bezafibrate hours are 15 – 45z, 35 – 65z and not less than 80z, respec-
is about 6 minutes. tively, and those of a 200-mg tablet in 1.5 hours, in 2.5 hours
Time span of measurement: About 2.5 times as long as the and in 8 hours are 15 – 45z, 30 – 60z and not less than
retention time of bezafibrate, beginning after the solvent 75z, respectively.
peak. Start the test with 1 tablet of Bezafibrate Extended-release
System suitability— Tablets, withdraw exactly 20 mL of the medium at the speci-
Test for required detectability: Measure exactly 5 mL of fied minutes after starting the test, and immediately fill
the standard solution, and add a mixture of methanol and up the dissolution medium each time with exactly 20 mL
diluted 0.5 mol/L ammonium acetate TS (1 in 50) (7:3) to of fresh dissolution medium, previously warmed to 37 ±
make exactly 50 mL. Confirm that the peak area of 0.59C. Filter these media through a membrane filter with a
bezafibrate obtained with 5 mL of this solution is equivalent pore size not exceeding 0.45 mm. Discard the first 10 mL of
to 7 to 13z of that obtained with 5 mL of the standard solu- the filtrate, pipet the subsequent V mL, add the dissolution
tion. medium to make exactly V? mL so that each mL contains
System performance: Dissolve 20 mg of Bezafibrate and about 13 mg of bezafibrate (C19H20ClNO4), and use these so-
10 mg of 4-chlorobenzoate in 70 mL of methanol, and add lutions as the sample solutions. Separately, weigh accurately
diluted 0.5 mol/L ammonium acetate TS (1 in 50) to make about 66 mg of bezafibrate for assay, previously dried at
100 mL. When the procedure is run with 5 mL of this solu- 1059C for 3 hours, and dissolve in methanol to make exactly
tion under the above operating conditions, 4-chlorobenzoate 50 mL. Pipet 2 mL of this solution, add the dissolution me-
and bezafibrate are eluted in this order with the resolution dium to make exactly 200 mL, and use this solution as the
between these peaks being not less than 3. standard solution. Determine the absorbances, AT(n) (n ＝
System repeatability: When the test is repeated 6 times 1,2,3) and AS, of the sample solutions and standard solution
with 5 mL of the standard solution under the above operating at 228 nm as directed under Ultraviolet-visible Spectropho-
conditions, the relative standard deviation of the peak area tometry <2.24>, using the dissolution medium as the blank.
of bezafibrate is not more than 2.0z.
Dissolution rate (z) in each case of n with respect to the
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, labeled amount of bezafibrate (C19H20ClNO4)
3 hours).
＝ MS ×
AT(n) ＋ n－1
 AS S i＝ 1
Ø AT(i)
AS
×
1
45 »
×
V? 1
V
× × 18
C
Assay Weigh accurately about 0.7 g of Bezafibrate, previ-
MS: Amount (mg) of bezafibrate for assay taken
ously dried, dissolve in 50 mL of ethanol (99.5), and titrate
C: Labeled amount (mg) of bezafibrate (C19H20ClNO4) in
<2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 3
1 tablet
drops of phenolphthalein TS). Perform a blank determina-
tion in the same manner, and make any necessary correction. Assay Weigh accurately, and powder not less than 20
Bezafibrate Extended-release Tablets. Weigh accurately a
portion of the powder, equivalent to about 20 mg of
bezafibrate (C19H20ClNO4), add 60 mL of methanol and
Containers and storage Containers—Tight containers. exactly 10 mL of the internal standard solution, and shake
for 20 minutes. Add diluted 0.5 mol/L ammonium acetate
JP XVII Official Monographs / Bifonazole 509
TS (1 in 50) to make 100 mL, filter, and use the filtrate as the show optical rotation.
of bezafibrate for assay, previously dried at 1059C for 3
solution of Bifonazole in methanol (1 in 100,000) as directed
hours, dissolve in 60 mL of methanol, add exactly 10 mL of
the internal standard solution and diluted 0.5 mol/L ammo-
nium acetate TS (1 in 50) to make 100 mL, and use this solu-
tion as the standard solution. Perform the test with 2 mL
wavelengths.
(2) Determine the infrared absorption spectrum of Bi-
fonazole, previously dried, as directed in the potassium bro-
the peak area of bezafibrate to that of the internal standard.
Amount (mg) of bezafibrate (C19H20ClNO4) ＝ MS × QT/QS both spectra exhibit similar intensities of absorption at the
same wave numbers.
MS: Amount (mg) of bezafibrate for assay taken
Internal standard solution—A solution of 4-nitrophenol in
methanol (1 in 500). Purity (1) Chloride <1.03>—To 2.0 g of Bifonazole add
Operating conditions— 40 mL of water, warm for 5 minutes, and after cooling,
Detector: An ultraviolet absorption photometer (wave- filter. To 10 mL of the filtrate add 6 mL of dilute nitric acid
length: 230 nm). and water to make 50 mL. Perform the test using this solu-
Column: A stainless steel column 4.6 mm in inside diame- tion as the test solution. Prepare the control solution with
ter and 15 cm in length, packed with octadecylsilanized silica 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than
gel for liquid chromatography (5 mm in particle diameter). 0.021z).
Column temperature: A constant temperature of about (2) Sulfate <1.14>—To 10 mL of the filtrate obtained in
259 C. (1) add 1 mL of dilute hydrochloric acid and water to make
Mobile phase: A mixture of methanol and diluted acetic 50 mL. Perform the test using this solution as the test solu-
acid (100) (1 in 100) (9:4). tion. Prepare the control solution with 0.50 mL of 0.005
Flow rate: Adjust so that the retention time of bezafibrate mol/L sulfuric acid VS (0.048z).
is about 6 minutes. (3) Heavy metals <1.07>—Proceed with 2.0 g of Bifona-
System suitability— zole according to Method 2, and perform the test. Prepare
System performance: When the procedure is run with 2 mL the control solution with 2.0 mL of Standard Lead Solution
of the standard solution under the above operating condi- (not more than 10 ppm).
tions, the internal standard and bezafibrate are eluted in this (4) Related substances—Conduct this procedure without
order with the resolution between these peaks being not less exposure to light, using light-resistant vessels. Dissolve 0.10
than 4. g of Bifonazole in 10 mL of methanol, and use this solution
System repeatability: When the test is repeated 6 times as the sample solution. Pipet 3 mL of the sample solution,
with 2 mL of the standard solution under the above operating and add methanol to make exactly 100 mL. Pipet 25 mL and
conditions, the relative standard deviation of the ratio of the 5 mL of this solution, add methanol to make exactly 50 mL
peak area of bezafibrate to that of the internal standard is each, and use these solutions as the standard solutions (1)
not more than 1.0z. and (2), respectively. Perform the test with these solutions as
mL each of the sample solution and standard solutions (1)
and (2) on a plate of silica gel with fluorescent indicator for
Bifonazole of ethyl acetate and ammonia solution (28) (49:1) to a dis-
ビホナゾール
ultraviolet light (main wavelength: 254 nm): the spot with R f
value of about 0.20 from the sample solution is not more in-
tense than the spot from the standard solution (1). And the
spots other than the spot mentioned above and the principal
spot from the standard solution (2).
C22H18N2: 310.39
1-[(RS )-(Biphenyl-4-yl)(phenyl)methyl]-1H-imidazole
[60628-96-8] Assay Weigh accurately about 0.15 g of Bifonazole, previ-
ously dried, and dissolve in dichloromethane to make exactly
Bifonazole, when dried, contains not less than 50 mL. Pipet 5 mL of this solution in a glass-stoppered coni-
98.5z of bifonazole (C22H18N2). cal flask, add 10 mL of water, 5 mL of dilute sulfuric acid
and 25 mL of dichloromethane, and add 2 to 3 drops of a
Description Bifonazole occurs as a white to pale yellow
solution of methyl yellow in dichloromethane (1 in 500)
powder. It is odorless and tasteless.
as indicator, and titrate <2.50>, while shaking vigorously,
It is freely soluble in dichloromethane, soluble in metha-
with 0.01 mol/L sodium lauryl sulfate VS by a buret with
nol, sparingly soluble in ethanol (95), slightly soluble in
0.02-mL minimum graduation. The end point is reached
diethyl ether, and practically insoluble in water.
when the color of the dichloromethane layer changes from
A solution of Bifonazole in methanol (1 in 100) does not
yellow to orange-red after dropwise addition of 0.01 mol/L
510 Biotin / Official Monographs JP XVII
sodium lauryl sulfate VS, strong shaking, and standing for a layer chromatography. Develop the plate with a mixture of
while. 1-butanol, water, and acetic acid (100) (5:2:1) to a distance
of about 10 cm, air-dry the plate, and then dry for 30
Each mL of 0.01 mol/L sodium lauryl sulfate VS
minutes at 1059C. Spray the plate evenly with a mixture of a
＝ 3.104 mg of C22H18N2
solution of 4-dimethylaminocinnamaldehyde in ethanol
Containers and storage Containers—Tight containers. (99.5) (1 in 500) and a solution of sulfuric acid in ethanol
Storage—Light-resistant. (99.5) (1 in 50) (1:1): the spots other than the principal spot
obtained from the sample solution are not more intense than
the spot obtained from the standard solution.
Biotin Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C,
4 hours).
ビオチン
Assay Weigh accurately about 0.25 g of Biotin, previously
dried, dissolve by adding exactly 20 mL of 0.1 mol/L sodium
hydroxide VS, and titrate <2.50> the excess sodium hydrox-
ide with 0.1 mol/L hydrochloric acid VS (indicator: 2 drops
C10H16N2O3S: 244.31 of phenolphthalein TS). Perform a blank determination in
5-[(3aS,4S,6aR)-2-Oxohexahydro-1H- the same manner.
thieno[3,4-d ]imidazol-4-yl]pentanoic acid
[58-85-5]
＝ 24.43 mg of C10H16N2O3S
Biotin, when dried, contains not less than 98.5z Containers and storage Containers—Tight containers.
and not more than 101.0z of biotin (C10H16N2O3S).
Description Biotin occurs as white crystals or a white crys-
talline powder. Biperiden Hydrochloride
It is very slightly soluble in water and in ethanol (99.5).
ビペリデン塩酸塩
It dissolves in dilute sodium hydroxide TS.
of Biotin as directed in the potassium bromide disc method
C21H29NO.HCl: 347.92
1-(Bicyclo[2.2.1]hept-5-en-2-yl)-1-phenyl-3-(piperidin-1-
0.4 g, dilute sodium hydroxide TS, 20 mL, 100 mm).
yl)propan-1-ol monohydrochloride
Purity (1) Clarity and color of solution—Dissolve 1.0 g [1235-82-1]
of Biotin in 10 mL of 0.5 mol/L sodium hydroxide TS: the
solution is clear and colorless. Biperiden Hydrochloride, when dried, contains
(2) Heavy metals <1.07>—Proceed with 2.0 g of Biotin not less than 99.0z of biperiden hydrochloride
according to Method 2, and perform the test. Prepare the (C21H29NO.HCl).
Description Biperiden Hydrochloride occurs as a white to
more than 10 ppm).
brownish and yellowish white crystalline powder.
(3) Arsenic <1.11>—Place 0.7 g of Biotin in a Kjeldahl
It is freely soluble in formic acid, slightly soluble in water,
flask, add 5 mL of nitric acid and 2 mL of sulfuric acid,
in methanol and in ethanol (95), and practically insoluble in
place a small funnel on the mouth of the flask, and carefully
diethyl ether.
heat until white fumes are evolved. After cooling, add 2 mL
of nitric acid twice, heat, add 2 mL of hydrogen peroxide
(30) several times, and heat until the color of the solution Identification (1) Dissolve 0.02 g of Biperiden Hydro-
becomes colorless or pale yellow. After cooling, add 2 mL of chloride in 5 mL of phosphoric acid: a green color develops.
saturated ammonium oxalate solution, and heat to concen- (2) Dissolve 0.01 g of Biperiden Hydrochloride in 5 mL
trate until white fumes are evolved again. After cooling, add of water by heating, cool, and add 5 to 6 drops of bromine
water to make 5 mL, and perform the test using this solution TS: a yellow precipitate is formed.
as the test solution (not more than 2.8 ppm). (3) Determine the absorption spectrum of a solution of
(4) Related substances—Dissolve 0.10 g of Biotin in 10 Biperiden Hydrochloride (1 in 2000) as directed under Ultra-
mL of diluted ammonia solution (28) (7 in 100), and use this violet-visible Spectrophotometry <2.24>, and compare the
solution as the sample solution. Pipet 1 mL of the sample spectrum with the Reference Spectrum: both spectra exhibit
solution, and add diluted ammonia solution (28) (7 in 100) similar intensities of absorption at the same wavelengths.
to make exactly 100 mL. Pipet 10 mL of this solution, add (4) Determine the infrared absorption spectrum of
diluted ammonia solution (28) (7 in 100) to make exactly 50 Biperiden Hydrochloride, previously dried, as directed in the
mL, and use this solution as the standard solution. Perform potassium chloride disk method under Infrared Spectropho-
the test with these solutions as directed under Thin-layer tometry <2.25>, and compare the spectrum with the Refer-
Chromatography <2.03>. Spot 5 mL each of the sample solu- ence Spectrum: both spectra exhibit similar intensities of ab-
tion and standard solution on a plate of silica gel for thin- sorption at the same wave numbers.
JP XVII Official Monographs / Bisacodyl 511
(5) Dissolve 0.02 g of Biperiden Hydrochloride in 10 mL tically insoluble in water.

of water by heating, and cool: the solution responds to the It dissolves in dilute hydrochloric acid.
Qualitative Tests <1.09> for chloride.
Purity (1) Acidity or alkalinity—To 1.0 g of Biperiden solution of Bisacodyl in ethanol (95) (3 in 100,000) as di-
Hydrochloride add 50 mL of water, shake vigorously, filter, rected under Ultraviolet-visible Spectrophotometry <2.24>,
and to 20 mL of the filtrate add 1 drop of methyl red TS: no and compare the spectrum with the Reference Spectrum or
red to yellow color develops. the spectrum of a solution of Bisacodyl RS prepared in the
(2) Heavy metals <1.07>—Proceed with 1.0 g of same manner as the sample solution: both spectra exhibit
Biperiden Hydrochloride according to Method 2, and per- similar intensities of absorption at the same wavelengths.
form the test. Prepare the control solution with 2.0 mL of (2) Determine the infrared absorption spectrum of Bi-
Standard Lead Solution (not more than 20 ppm). sacodyl, previously dried, as directed in the potassium bro-
(3) Arsenic <1.11>—Prepare the test solution with 1.0 g mide disk method under Infrared Spectrophotometry <2.25>,
of Biperiden Hydrochloride according to Method 3, and per- and compare the spectrum with the Reference Spectrum or
form the test (not more than 2 ppm). the spectrum of dried Bisacodyl RS: both spectra exhibit
(4) Related substances—Dissolve 0.10 g of Biperiden Hy- similar intensities of absorption at the same wave numbers.
drochloride in 20 mL of methanol, and use this solution as
methanol to make exactly 200 mL, and use this solution as Purity (1) Chloride <1.03>—Dissolve 1.0 g of Bisacodyl in
the standard solution. Perform the test with these solutions 30 mL of acetone, and add 6 mL of dilute nitric acid and
as directed under Thin-layer Chromatography <2.03>. Spot water to make 50 mL. Perform the test using this solution as
50 mL each of the sample solution and standard solution on a the test solution. Prepare the control solution as follows: to
plate of silica gel for thin-layer chromatography. Develop 0.35 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of
the plate with a mixture of chloroform, methanol and am- acetone, 6 mL of dilute nitric acid and water to make 50 mL
monia solution (28) (80:15:2) to a distance of about 15 cm, (not more than 0.012z).
and air-dry the plate. Spray evenly Dragendorff's TS for (2) Sulfate <1.14>—Dissolve 1.0 g of Bisacodyl in 2 mL
spraying on the plate: the spots other than the principal spot of dilute hydrochloric acid, and add water to make 50 mL.
from the sample solution are not more intense than the spot Perform the test using this solution as the test solution. Pre-
from the standard solution. pare the control solution as follows: to 0.35 mL of 0.005
mol/L sulfuric acid VS add 2 mL of dilute hydrochloric acid
and water to make 50 mL (not more than 0.017z).
3 hours).
(3) Heavy metals <1.07>—Proceed with 2.0 g of Bi-
Residue on ignition <2.44> Not more than 0.1z (1 g). sacodyl according to Method 4, and perform the test. Pre-
Assay Weigh accurately about 0.4 g of Biperiden Hydro-
lution (not more than 10 ppm).
chloride, previously dried, dissolve in 5 mL of formic acid,
(4) Related substances—Dissolve 0.20 g of Bisacodyl in
10 mL of acetone, and use this solution as the sample solu-
tion. Pipet 1 mL of the sample solution, add acetone to
tion.
Each mL of 0.1 mol/L perchloric acid VS under Thin-layer Chromatography <2.03>. Spot 10 mL each
＝ 34.79 mg of C21H29NO.HCl of the sample solution and standard solution on a plate of
raphy. Develop the plate with a mixture of 2-butanone, chlo-
ers.
roform and xylene (1:1:1) to a distance of about 10 cm, and
Bisacodyl from the standard solution.
ビサコジル Loss on drying <2.41> Not more than 0.5z (1 g, 1059C,
2 hours).
Assay Weigh accurately about 0.5 g of Bisacodyl, previ-
<2.50> with 0.1 mol/L perchloric acid VS until the color of
the solution changes from orange-yellow to green (indicator:
C22H19NO4: 361.39 0.5 mL of p-naphtholbenzein TS). Perform a blank determi-
4,4?-(Pyridin-2-ylmethylene)bis(phenyl acetate) nation, and make any necessary correction.
[603-50-9]
＝ 36.14 mg of C22H19NO4
Bisacodyl, when dried, contains not less than 98.5z
of bisacodyl (C22H19NO4). Containers and storage Containers—Well-closed contain-
ers.
Description Bisacodyl occurs as a white crystalline powder.
It is freely soluble in acetic acid (100), soluble in acetone,
slightly soluble in ethanol (95) and in diethyl ether, and prac-
512 Bisacodyl Suppositories / Official Monographs JP XVII
Bisacodyl Suppositories conditions, and calculate the ratios, QT and QS, of the peak
area of bisacodyl to that of the internal standard.
ビサコジル坐剤
Amount (mg) of bisacodyl (C22H19NO4) ＝ MS × QT/QS
MS: Amount (mg) of Bisacodyl RS taken
Bisacodyl Suppositories contain not less than 90.0z
and not more than 110.0z of the labeled amount of Internal standard solution—A solution of ethyl parahy-
bisacodyl (C22H19NO4: 361.39). droxybenzoate in acetonitrile (3 in 100,000).
Method of preparation Prepare as directed under Supposi-
tories, with Bisacodyl.
length: 254 nm).
Identification (1) To a quantity of Bisacodyl Supposito- Column: A stainless steel column 4 mm in inside diameter
ries, equivalent to 6 mg of Bisacodyl, add 20 mL of ethanol and 30 cm in length, packed with octadecylsilanized silica gel
(95), warm on a water bath for 10 minutes, shake vigorously for liquid chromatography (10 mm in particle diameter).
for 10 minutes, and allow to stand in ice water for 1 hour. Column temperature: A constant temperature of about
Centrifuge the solution, filter the supernatant liquid, and to 259C.
2 mL of the filtrate add ethanol (95) to make 20 mL. Deter- Mobile phase: A mixture of 0.01 mol/L citric acid TS,
mine the absorption spectrum of the solution as directed acetonitrile and methanol (2:1:1).
under Ultraviolet-visible Spectrophotometry <2.24>: it exhib- Flow rate: Adjust so that the retention time of bisacodyl is
its a maximum between 261 nm and 265 nm. about 8 minutes.
(2) Use the filtrate obtained in (1) as the sample solution. System suitability—
Separately, dissolve 6 mg of Bisacodyl RS in 20 mL of System performance: When the procedure is run with 20
ethanol (95), and use this solution as the standard solution. mL of the standard solution under the above operating con-
Perform the test with these solutions as directed under Thin- ditions, the internal standard and bisacodyl are eluted in this
layer Chromatography <2.03>. Spot 20 mL each of the sample order with the resolution between these peaks being not less
solution and standard solution on a plate of silica gel with than 2.0.
fluorescent indicator for thin-layer chromatography. De- System repeatability: When the test is repeated 6 times
velop the plate with a mixture of 2-butanone, chloroform with 20 mL of the standard solution under the above operat-
and xylene (1:1:1) to a distance of about 10 cm, and air-dry ing conditions, the relative standard deviation of the ratios
the plate. Examine under ultraviolet light (main wavelength: of the peak area of bisacodyl to that of the internal standard
254 nm): the spot from the sample solution and that from is not more than 1.0z.
the standard solution show the same R f value.
Content uniformity test. Bismuth Subgallate
To 1 suppository of Bisacodyl Suppositories add a suitable
amount of tetrahydrofuran, warm to 409C, and shake to dis- Dermatol
solve. After cooling, add tetrahydrofuran to make exactly
V mL so that each mL contains about 0.2 mg of bisacodyl 次没食子酸ビスマス
(C22H19NO4). Pipet 5 mL of this solution, and proceed as
directed in the Assay.
Bismuth Subgallate, when dried, contains not less
Amount (mg) of bisacodyl (C22H19NO4) than 47.0z and not more than 51.0z of bismuth (Bi:
＝ MS × QT/QS × V/50 208.98).
MS: Amount (mg) of Bisacodyl RS taken Description Bismuth Subgallate occurs as a yellow powder.
It is odorless and tasteless.
Internal standard solution—A solution of ethyl parahy-
droxybenzoate in acetonitrile (3 in 100,000).
diethyl ether.
Assay Weigh accurately not less than 20 Bisacodyl Sup- It dissolves in dilute hydrochloric acid, in dilute nitric acid
positories, make them fine fragments carefully, and mix and in dilute sulfuric acid on warming. It dissolves in sodium
uniformly. Weigh accurately a portion of the fragments, hydroxide TS, forming a clear, yellow solution, which turns
equivalent to about 10 mg of bisacodyl (C22H19NO4), add 40 red immediately.
mL of tetrahydrofuran, warm to 409C, dissolve by shaking, It is affected by light.
cool, and add tetrahydrofuran to make exactly 50 mL. Pipet
Identification (1) Ignite 0.5 g of Bismuth Subgallate: it
5 mL of this solution, add exactly 5 mL of the internal stand-
chars at first, and leaves finally a yellow residue. The residue
ard solution, and add the mobile phase to make 100 mL.
responds to the Qualitative Tests <1.09> for bismuth salt.
Cool this solution in ice for 30 minutes, centrifuge, filter the
(2) To 0.5 g of Bismuth Subgallate add 25 mL of water
supernatant liquid through a membrane filter with pore size
and 20 mL of hydrogen sulfide TS, and shake well. Filter off
of 0.5 mm, discard the first 10 mL of the filtrate, and use the
the blackish brown precipitate, and add 1 drop of iron (III)
subsequent filtrate as the sample solution. Separately, weigh
chloride TS to the filtrate: a blue-black color is produced.
accurately about 10 mg of Bisacodyl RS, previously dried at
1059C for 2 hours, and dissolve in tetrahydrofuran to make Purity (1) Clarity of solution—Dissolve 1.0 g of Bismuth
exactly 50 mL. Pipet 5 mL of this solution, proceed in the Subgallate in 40 mL of diluted sodium hydroxide TS (1 in 8):
same manner as the sample solution, and use this solution as the solution is clear.
the standard solution. Perform the test with 20 mL each of (2) Sulfate—Ignite 3.0 g of Bismuth Subgallate in a por-
the sample solution and standard solution as directed under celain crucible, and cautiously dissolve the residue in 2.5 mL
JP XVII Official Monographs / Bismuth Subnitrate 513
of nitric acid by warming. Pour the solution into 100 mL of Storage—Light-resistant.

water, shake, and filter. Evaporate 50 mL of the filtrate on a
water bath to 15 mL. Add water to make 20 mL, filter again,
and use the filtrate as the sample solution. To 5 mL of the Bismuth Subnitrate
sample solution add 2 to 3 drops of barium nitrate TS: no
turbidity is produced. 次硝酸ビスマス
(3) Nitrate—To 0.5 g of Bismuth Subgallate add 5 mL of
dilute sulfuric acid and 25 mL of iron (II) sulfate TS, shake
Bismuth Subnitrate, when dried, contains not less
well, and filter. Superimpose carefully 5 mL of the filtrate
than 71.5z and not more than 74.5z of bismuth (Bi:
on sulfuric acid: no red-brown color develops at the zone of
208.98).
contact.
(4) Ammonium—Dissolve 1.0 g of Bismuth Subgallate in Description Bismuth Subnitrate occurs as a white powder.
5 mL of sodium hydroxide TS, and heat: the gas evolved It is practically insoluble in water, in ethanol (95) and in
does not change moistened red litmus paper to blue. diethyl ether.
(5) Copper—To 5 mL of the sample solution obtained in It readily dissolves in hydrochloric acid and in nitric acid
(2) add 1 mL of ammonia TS, and filter: no blue color de- without effervescence.
velops in the filtrate. It is slightly hygroscopic, and changes moistened blue
(6) Lead—Ignite 1.0 g of Bismuth Subgallate at about litmus paper to red.
5009C in a porcelain crucible, dissolve the residue in a
Identification Bismuth Subnitrate responds to the Qualita-
smallest possible amount of nitric acid added dropwise,
tive Tests <1.09> for bismuth salt and nitrate.
evaporate over a low flame to dryness , and cool. Add 5 mL
of a solution of potassium hydroxide (1 in 6) to the residue, Purity (1) Chloride <1.03>—Dissolve 0.7 g of Bismuth
boil carefully for 2 minutes, cool, and centrifuge. Take the Subnitrate in 2 mL of water and 2 mL of nitric acid, and add
supernatant liquid in a test tube, add 10 drops of potassium 6 mL of dilute nitric acid and water to make 50 mL. Perform
chromate TS, and acidify the solution by adding acetic acid the test using this solution as the test solution. Prepare the
(100) dropwise: neither turbidity nor a yellow precipitate is control solution as follows: evaporate 2 mL of nitric acid on
produced. a water bath to dryness, add 0.70 mL of 0.01 mol/L hydro-
(7) Silver—To 5 mL of the sample solution obtained in chloric acid VS, 6 mL of dilute nitric acid and water to make
(2) add 0.5 mL of nitric acid and 2 to 3 drops of dilute hy- 50 mL (not more than 0.035z).
drochloric acid: no turbidity is produced. (2) Sulfate—Dissolve 3.0 g of Bismuth Subnitrate in 3.0
(8) Alkaline earth metals and alkali metals—Boil 1.0 g of mL of warmed nitric acid, pour this solution into 100 mL of
Bismuth Subgallate with 40 mL of diluted acetic acid (31) (1 water, shake, and filter. Concentrate the filtrate on a water
in 2) for 2 minutes, cool, add water to make 40 mL, and bath to 30 mL, filter, and use this filtrate as the sample solu-
filter. To 20 mL of the filtrate add 2 mL of dilute hydrochlo- tion. To 5 mL of the sample solution add 2 to 3 drops of
ric acid, boil, immediately pass hydrogen sulfide thoroughly barium nitrate TS: no turbidity is produced.
through the solution, filter the precipitate produced, and (3) Ammonium—Boil 0.10 g of bismuth Subnitrate with
wash with water. Combine the filtrate and the washings, add 5 mL of sodium hydroxide TS: the gas evolved does not
5 drops of sulfuric acid, and evaporate to dryness. Ignite as change moistened red litmus paper to blue.
directed under Residue on Ignition <2.44>: the mass of the (4) Copper—To 5 mL of the sample solution obtained in
residue is not more than 5.0 mg. (2) add 2 mL of ammonia TS, and filter: no blue color de-
(9) Arsenic <1.11>—Mix well 0.20 g of Bismuth Subgal- velops.
late with 0.20 g of calcium hydroxide, and ignite the mixture. (5) Lead—To 1.0 g of Bismuth Subnitrate add 5 mL of a
Dissolve the residue in 5 mL of dilute hydrochloric acid, use solution of sodium hydroxide (1 in 6), boil carefully for 2
this solution as the test solution, and perform the test (not minutes, cool and centrifuge. Transfer the supernatant liq-
more than 10 ppm). uid to a test tube, add 10 drops of potassium chromate TS,
(10) Gallic acid—To 1.0 g of Bismuth Subgallate add and add dropwise acetic acid (31) to render the solution acid:
20 mL of ethanol (95), shake for 1 minute, and filter. no turbidity or yellow precipitate is produced.
Evaporate the filtrate on a water bath to dryness: the mass (6) Silver—To 5 mL of the sample solution obtained in
of the residue is not more than 5.0 mg. (2) add 0.5 mL of nitric acid and 2 to 3 drops of dilute hy-
drochloric acid: no turbidity is produced.
(7) Alkaline earth metals and alkali metals—Boil 2.0 g of
3 hours).
Bismuth Subnitrate with 40 mL of diluted acetic acid (31) (1
Assay Weigh accurately about 0.5 g of Bismuth Subgallate, in 2) for 2 minutes, cool, add water to make 40 mL, and
previously dried, ignite at about 5009C for 30 minutes, and filter. To 20 mL of the filtrate add 2 mL of dilute hydrochlo-
cool. Dissolve the residue in 5 mL of diluted nitric acid (2 in ric acid, boil, immediately pass hydrogen sulfide thoroughly
5) by warming, and add water to make exactly 100 mL. through the solution, filter, and wash the residue with water.
Measure exactly 30 mL of this solution, add 200 mL of Combine the filtrate and the washings, add 5 drops of sulfu-
water, and titrate <2.50> with 0.02 mol/L disodium dihydro- ric acid, evaporate to dryness, and ignite as directed under
gen ethylenediamine tetraacetate VS until the color of the so- Residue on Ignition <2.44>: the residue is not exceed 5.0 mg
lution changes from red-purple to yellow (indicator: 2 to 3 (8) Arsenic <1.11>—To 0.20 g of Bismuth Subnitrate add
drops of xylenol orange TS). 2 mL of sulfuric acid, heat until white fumes evolve, dilute
cautiously with water to 5 mL, use this solution as the test
solution, and perform the test (not more than 10 ppm).
＝ 4.180 mg of Bi Loss on drying <2.41> Not more than 3.0z (2 g, 1059C,
2 hours).
ers. Assay Weigh accurately about 0.4 g of Bismuth Subnitrate,
514 Bisoprolol Fumarate / Official Monographs JP XVII
previously dried, dissolve in 5 mL of diluted nitric acid (2 in the following conditions. Determine each peak area of both
5) by warming, and add water to make exactly 100 mL. Pipet solutions by the automatic integration method: the area of
25 mL of the solution, add 200 mL of water and titrate the peaks other than bisoprolol obtained from the sample so-
<2.50> with 0.02 mol/L disodium dihydrogen ethylene- lution is not larger than 1/2 times the peak area of bisoprolol
diamine tetraacetate VS until the color of the solution obtained from the standard solution. Furthermore, the total
changes from red-purple to yellow (indicator: 5 drops of of the areas of peaks other than bisoprolol from the sample
xylenol orange TS) solution is not larger than the peak area of bisoprolol from
＝ 4.180 mg of Bi
length: 225 nm).
Containers and storage Containers—Well-closed contain- Column: A stainless steel column 4.6 mm in inside diame-
ers. ter and 15 cm in length, packed with octylsilanized silica gel
Bisoprolol Fumarate 409C.
ビソプロロールフマル酸塩 phosphate in 1000 mL of water, and adjust to pH 2.5 with
phosphoric acid. To 800 mL of this solution add 200 mL of
acetonitrile.
Flow rate: Adjust so that the retention time of bisoprolol
is about 8 minutes.
retention time of bisoprolol, beginning after the fumaric acid
peak.
(C18H31NO4)2.C4H4O4: 766.96 Test for required detectability: Pipet 2 mL of the standard
(2RS )-1-(4-{[2-(1-Methylethoxy)ethoxy]methyl}phenoxy)- solution, and add a mixture of water and acetonitrile (4:1)
3-[(1-methylethyl)amino]propan-2-ol hemifumarate to make exactly 20 mL. Confirm that the peak area of bi-
[104344-23-2] soprolol obtained from 20 mL of this solution is equivalent to
7 to 13z of that obtained from 20 mL of the standard solu-
Bisoprolol Fumarate, when dried, contains not tion.
less than 98.5z and not more than 101.0z of System performance: When the procedure is run with 20
bisoprolol fumarate [(C18H31NO4)2.C4H4O4]. mL of the standard solution under the above operating con-
Description Bisoprolol Fumarate occurs as white crystals
factor of the peak of bisoprolol are not less than 5000 and
or a white crystalline powder.
It is very soluble in water and in methanol, and freely
soluble in ethanol (99.5) and in acetic acid (100).
A solution of Bisoprolol Fumarate (1 in 10) shows no opti-
cal rotation.
area of bisoprolol is not more than 1.5z.
solution of Bisoprolol Fumarate in methanol (1 in 10,000) as
um, phosphorus (V) oxide, 809
C, 5 hours).
and compare the spectrum with the Reference Spectrum: Residue on ignition <2.44> Not more than 0.1z (1 g).
Assay Weigh accurately about 0.6 g of Bisoprolol
same wavelengths.
Fumarate, previously dried, dissolve in 70 mL of acetic acid
(2) Determine the infrared absorption spectrum of Bi-
(100), and titrate <2.50> with 0.1 mol/L perchloric acid VS
soprolol Fumarate as directed in the potassium bromide disc
(indicator: 2 drops of crystal violet TS). The endpoint of
titration is when the purple color of the solution turns blue
and then blue-green. Perform a blank determination in the
numbers.
＝ 38.35 mg of (C18H31NO4)2.C4H4O4
Bisoprolol Fumarate according to Method 2, and perform
(2) Related substances—Dissolve 50 mg of Bisoprolol
Fumarate in 100 mL of a mixture of water and acetonitrile
mL of the sample solution, add the mixture of water and
acetonitrile (4:1) to make exactly 100 mL, and use this solu-
JP XVII Official Monographs / Bisoprolol Fumarate Tablets 515

Bisoprolol Fumarate Tablets with 20 mL of the solution for system suitability test under
ビソプロロールフマル酸塩錠 tion of the peak area of bisoprolol is not more than 1.5z.
Bisoprolol Fumarate Tablets contain not less than ing to the following method: it meets the requirement of the
95.0z and not more than 105.0z of the labeled Content uniformity test.
amount of bisoprolol fumarate [(C18H31NO4)2.C4H4O4: Take 1 tablet of Bisoprolol Fumarate Tablets, disintegrate
766.96]. by adding 8 mL of water, and add water to make exactly 10
mL, and then filter through a membrane filter with a pore
size not exceeding 0.45 mm. Discard the first 3 mL of the fil-
with Bisoprolol Fumarate.
trate, pipet V mL of the subsequent filtrate, add water to
Identification To a quantity of powdered Bisoprolol make exactly V? mL so that each mL contains about 62.5 mg
Fumarate Tablets, equivalent to 10 mg of Bisoprolol of bisoprolol fumarate [(C18H31NO4)2.C4H4O4], and use this
Fumarate, add 60 mL of methanol, shake vigorously for 10 solution as the sample solution. Separately, weigh accurately
minutes, add methanol to make 100 mL, and filter through a about 20 mg of bisoprolol fumarate for assay, previously
membrane filter with a pore size not exceeding 0.45 mm. De- dried under reduced pressure at 809C for 5 hours, using
termine the absorption spectrum of the filtrate as directed phosphorus (V) oxide as a dessicant, and dissolve in water to
under Ultraviolet-visible Spectrophotometry <2.24>: it exhib- make exactly 200 mL. Pipet 15 mL of this solution, add
its a maximum between 271 nm and 275 nm. water to make exactly 25 mL, and use this solution as the
standard solution. Determine the absorbances, AT and AS, at
Purity Related substances—This is applied to 0.625-mg
271.5 nm of the sample solution and standard solution as
tablets. Shake vigorously for 10 minutes a portion of pow-
directed under Ultraviolet-visible Spectrophotometry <2.24>.
dered Bisoprolol Fumarate Tablets, equivalent to 5 mg of Bi-
soprolol Fumarate, with exactly 20 mL of a mixture of water Amount (mg) of bisoprolol fumarate
and acetonitrile (3:1), filter through a membrane filter with a [(C18H31NO4)2.C4H4O4]
pore size not exceeding 0.45 mm. Discard the first 3 mL of ＝ MS × AT/AS × V?/V × 3/100
the filtrate, and use the subsequent filtrate as the sample so-
MS: Amount (mg) of bisoprolol fumarate for assay taken
lution. Perform the test with 20 mL of the sample solution as
directed under Liquid Chromatography <2.01> according to Dissolution <6.10> When the test is performed at 50 revolu-
the following conditions. Determine each peak area by the tions per minute according to the Paddle method, using 900
automatic integration method, and calculate the amount of mL of 2nd fluid for dissolution test as the dissolution me-
the peak other than bisoprolol and the peak having the rela- dium, the dissolution rate in 30 minutes of Bisoprolol
tive retention time of about 0.8 to bisoprolol by the area per- Fumarate Tablets is not less than 85z.
centage method: the amount of the two peaks, having rela- Start the test with 1 tablet of Bisoprolol Fumarate Tablets,
tive retention time of about 1.2 and about 3.8 to bisoprolol, withdraw not less than 20 mL of the medium at the specified
are not more than 1.0z, respectively, the amount of the minute after starting the test, and filter through a membrane
peak other than the peaks mentioned above is not more than filter with a pore size not exceeding 0.45 mm. Discard the
0.2z, and the total amount of the peaks other than bi- first 10 mL of the filtrate, pipet V mL of the subsequent fil-
soprolol is not more than 2.5z. For the area of the peak, trate, add the dissolution medium to make exactly V? mL so
having the relative retention time of about 1.2 to bisoprolol, that each mL contains about 0.7 mg of bisoprolol fumarate
multiply the relative response factor 5. [(C18H31NO4)2.C4H4O4], and use this solution as the sample
Operating conditions— solution. Separately, weigh accurately about 14 mg of bi-
Detector, column, column temperature, and flow rate: soprolol fumarate for assay, previously dried in vacuum at
Proceed as directed in the operating conditions in the Assay. 809C for 5 hours, using phosphorus (V) oxide as a dessicant,
Mobile phase: Dissolve 4.08 g of potassium dihydrogen and dissolve in the dissolution medium to make exactly 100
phosphate in water to make 1000 mL, and adjust to pH 2.5 mL. Pipet 1 mL of this solution, add the dissolution medium
with phosphoric acid. To 750 mL of this solution add 250 to make exactly 200 mL, and use this solution as the stand-
mL of acetonitrile. ard solution. Perform the test with exactly 50 mL each of the
Time span of measurement: About 5 times as long as the sample solution and standard solution as directed under Liq-
retention time of bisoprolol, beginning after the peak of uid Chromatography <2.01>, and determine the peak areas,
fumaric acid. AT and AS, of bisoprolol in each solution.
of bisoprolol fumarate [(C18H31NO4)2.C4H4O4]
lution add a mixture of water and acetonitrile (3:1) to make
＝ MS × AT/AS × V?/V × 1/C × 9/2
100 mL, and use this solution as the solution for system
suitability test. Pipet 2 mL of the solution for system suita- MS: Amount (mg) of bisoprolol fumarate for assay taken
bility test, and add a mixture of water and acetonitrile (3:1) C: Labeled amount (mg) of bisoprolol fumarate
to make exactly 20 mL. Confirm that the peak area of bi- [(C18H31NO4)2.C4H4O4] in 1 tablet
soprolol obtained with 20 mL of this solution is equivalent to
7 to 13z of that obtained with 20 mL of the solution for sys-
Detector, column, column temperature, and flow rate:
tem suitability test.
Proceed as directed in the operating conditions in the Assay.
phosphate in 1000 mL of water, and adjust to pH 2.5 with
phosphoric acid. To 750 mL of this solution add 250 mL of
the symmetry factor of the peak of bisoprolol are not less
acetonitrile.
than 5000 and not more than 2.0, respectively.
516 Bleomycin Hydrochloride / Official Monographs JP XVII
System performance: When the procedure is run with 50 Bleomycin Hydrochloride
ditions, the number of theoretical plates and the symmetry ブレオマイシン塩酸塩
factor of the peak of bisoprolol are not less than 3000 and
area of bisoprolol is not more than 2.0z.
Assay Weigh accurately not less than 20 Bisoprolol
Fumarate Tablets and powder. Weigh accurately a portion
of the powder, equivalent to about 20 mg of bisoprolol
fumarate [(C18H31NO4)2.C4H4O4], add 70 mL of a mixture of
water and acetonitrile (3:1) and exactly 10 mL of the internal
standard solution, shake vigorously for 10 minutes, and add
the mixture of water and acetonitrile (3:1) to make 100 mL.
Filter this solution through a membrane filter with a pore
size not exceeding 0.45 mm, discard the first 3 mL of the fil-
trate, and use the subsequent filtrate as the sample solution.
Separately, weigh accurately about 20 mg of bisoprolol
fumarate for assay, previously dried in vacuum at 809C for
5 hours using phosphorus (V) oxide as the dessicant, add ex-
actly 10 mL of the internal standard solution, dissolve in the
mixture of water and acetonitrile (3:1) to make 100 mL, and
QS, of the peak area of bisoprolol to that of the internal
standard.
Amount (mg) of bisoprolol fumarate [(C18H31NO4)2.C4H4O4]
＝ MS × QT/QS
Bleomycinoic Acid
MS: Amount (mg) of bisoprolol fumarate for assay taken
1-Bleomycinoic acid hydrochloride
Internal standard solution—A solution of isopropyl parahy- Bleomycin A1
droxybenzoate in the mixture of water and acetonitrile (3:1) N 1-[3-(Methylsulfinyl)propyl]bleomycinamide
(1 in 250). hydrochloride
Operating conditions— Bleomycin Demethyl-A2
Detector: An ultraviolet absorption photometer (wave- N 1-[3-(Methylsulfanyl)propyl]bleomycinamide
length: 225 nm). hydrochloride
Column: A stainless steel column 4.6 mm in inside diame- Bleomycin A2
ter and 15 cm in length, packed with octylsilanized silica gel N 1-[3-(Dimethylsulfonio)propyl]bleomycinamide
for liquid chromatography (5 mm in particle diameter). hydrochloride
Column temperature: A constant temperature of about Bleomycin A2?-a
409 C. N 1-(4-Aminobutyl)bleomycinamide hydrochloride
Mobile phase: Dissolve 4.08 g of potassium dihydrogen Bleomycin A2?-b
phosphate in 1000 mL of water, and adjust to pH 2.5 with N 1-(3-Aminopropyl)bleomycinamide hydrochloride
phosphoric acid. To 800 mL of this solution add 200 mL of Bleomycin A5
acetonitrile. N 1-{3-[(4-Aminobutyl)amino]propyl}bleomycinamide hy-
Flow rate: Adjust so that the retention time of bisoprolol drochloride
is about 8 minutes. Bleomycin B1?
System suitability— Bleomycinamide hydrochloride
System performance: When the procedure is run with 20 Bleomycin B2
mL of the standard solution under the above operating con- N 1-(4-Guanidinobutyl)bleomycinamide
ditions, fumaric acid, bisoprolol and the internal standard hydrochloride
are eluted in this order with the resolution between the peaks Bleomycin B4
of bisoprolol and the internal standard being not less than N 1-{4-[3-(4-Guanidinobutyl)guanidino]butyl}-
12. bleomycinamide hydrochloride
System repeatability: When the test is repeated 6 times [11056-06-7, Bleomycin]
ing conditions, the relative standard deviation of the ratio of Bleomycin Hydrochloride is the hydrochloride of a
the peak area of bisoprolol to that of the internal standard is mixture of substances having antitumor activity pro-
not more than 1.0z. duced by the growth of Streptomyces verticillus.
more than 2000 mg (potency) per mg, calculated on
JP XVII Official Monographs / Bleomycin Hydrochloride 517
the dried basis. The potency of Bleomycin Hydrochlo-

ride is expressed as mass (potency) of bleomycin Time after injection Mobile phase A Mobile phase B
A2 (C55H84ClN17O21S3: 1451.00). of sample (min) (volz) (volz)
Description Bleomycin Hydrochloride occurs as a white to 0 – 60 100 → 0 0 → 100

yellowish white powder. 60 – 75 0 100
It is freely soluble in water, and slightly soluble in ethanol
(95). Flow rate: About 1.2 mL per minute.
It is hygroscopic. Time span of measurement: 20 minutes after elution of the
Identification (1) To 4 mg of Bleomycin Hydrochloride peak of demethylbreomycin A2, beginning after the solvent
add 5 mL of copper (II) sulfate TS, and dissolve in water to peak.
make 100 mL. Determine the absorption spectrum of this so- System suitability—
lution as directed under Ultraviolet-visible Spectrophotome- System performance: When the procedure is run with 20
try <2.24>, and compare the spectrum with the Reference mL of the sample solution under the above operating condi-
Spectrum: both spectra exhibit similar intensities of absorp- tions, bleomycin A2 and bleomycin B2 are eluted in this order
tion at the same wavelengths. with the resolution between these peaks being not less than 5.
(2) Determine the infrared absorption spectrum of System repeatability: When the test is repeated 6 times
Bleomycin Hydrochloride as directed in the potassium bro- with 20 mL of the sample solution under the above operating
mide disk method under Infrared Spectrophotometry <2.25>, conditions, the relative standard deviation of the peak area
and compare the spectrum with the Reference Spectrum: of bleomycin A2 is not more than 2.0z.
both spectra exhibit similar intensities of absorption at the Purity (1) Clarity and color of solution—A solution ob-
same wave numbers. tained by dissolving 80 mg of Bleomycin Hydrochloride in 4
(3) A solution of Bleomycin Hydrochloride (1 in 100) re- mL of water is clear and colorless.
sponds to the Qualitative Tests <1.09> (2) for chloride. (2) Copper—Dissolve exactly 75 mg of Bleomycin Hy-
pH <2.54> The pH of a solution obtained by dissolving drochloride in exactly 10 mL of diluted nitric acid (1 in 100),
0.10 g of Bleomycin Hydrochloride in 20 mL of water is be- and use this solution as the sample solution. Separately, to
tween 4.5 and 6.0. exactly 15 mL of Standard Copper Solution add diluted
nitric acid (1 in 100) to make exactly 100 mL, and use this
Content ratio of the active principle Dissolve 10 mg of solution as the standard solution. Perform the test with the
Bleomycin Hydrochloride in 20 mL of water, and use this so- sample solution and standard solution as directed under
lution as the sample solution. Perform the test with 20 mL of Atomic Absorption Spectrophotometry <2.23> according to
the sample solution as directed under Liquid Chromatogra- the following conditions: the absorbance of the sample solu-
phy <2.01> according to the following conditions, and deter- tion is not more than that of the standard solution (not more
mine each peak area by the automatic integration method, than 200 ppm).
and calculate their amounts by the area percentage method: Gas: Combustible gas—Acetylene.
the amount of the peak of bleomycin A2 (the first principal Supporting gas—Air.
peak) is between 55z and 70z, that of bleomycin B2 (the Lamp: Copper hollow-cathode lamp.
second principal peak) is between 25z and 32z, the total Wavelength: 324.8 nm.
amount of the peak of bleomycin A2 and bleomycin B2 is not
less than 85z, the amount of the peak of demethylbleomy- Loss on drying <2.41> Not more than 5.0z (60 mg, in
cin A2 (a peak having the relative retention time of 1.5 – 2.5 vacuum, phosphorus (V) oxide, 609 C, 3 hours. Take the
to bleomycin A2) is not more than 5.5z, and the total sample to be tested while avoiding moisture absorption).
amount of the rest peaks is not more than 9.5z. Assay Perform the test according to the Cylinder-plate
Operating conditions— method as directed under Microbial Assay for Antibiotics
Detector: An ultraviolet absorption photometer (wave- <4.02> according to the following conditions.
length: 254 nm). (i) Test organism—Mycobacterium smegmatis ATCC
Column: A stainless steel column 4.6 mm in inside diame- 607
ter and 25 cm in length, packed with octadecylsilanized silica (ii) Agar medium for seed, base layer and transferring
gel for liquid chromatography (7 mm in particle diameter). the test organism
Column temperature: A constant temperature of about Glycerin 10.0 g
409 C. Peptone 10.0 g
Mobile phase stock solution: Dissolve 0.96 g of sodium 1- Meat extract 10.0 g
pentanesulfonate and 1.86 g of disodium dihydrogen ethyl- Sodium chloride 3.0 g
enediamine tetraacetate dihydrate in 1000 mL of water and Agar 15.0 g
5 mL of acetic acid (100), and adjust the pH to 4.3 with Water 1000 mL
ammonia TS. Mix all the components, and sterilize. Adjust the pH after
Mobile phase A: A mixture of the mobile phase stock so- sterilization to 6.9 – 7.1 with sodium hydroxide TS.
lution and methanol (9:1). (iii) Liquid media for suspending the test organism
Mobile phase B: A mixture of the mobile phase stock solu- Glycerin 10.0 g
tion and methanol (3:2). Peptone 10.0 g
Flowing of mobile phase: Control the gradient by mixing Meat extract 10.0 g
the mobile phases A and B as directed in the following table. Sodium chloride 3.0 g
Water 1000 mL
Mix all the components, and sterilize. Adjust the pH after
sterilization to 6.9 – 7.1 with sodium hydroxide TS.
(iv) Preparation of seeded agar layer—Cultivate the test
518 Bleomycin Sulfate / Official Monographs JP XVII
organism on the slant of the agar medium for transferring
the test organism at 279C for 40 to 48 hours, then inoculate Bleomycin Sulfate
the test organism thus obtained in 100 mL of the liquid me-
dia for suspending the test organism, cultivate with shaking ブレオマイシン硫酸塩
at between 259C and 279C for 5 days, and use this as the sus-
pension of test organism. Store the suspension of test organ-
ism at a temperature not exceeding 59C, and use within 14
days. Add 0.5 mL of the suspension of test organism in 100
mL of the agar medium for seed previously kept at 489C,
mix thoroughly, and use as the seeded agar layer.
(v) Preparation of cylinder-agar plate—Proceed as di-
rected in 1.7. Preparation of cylinder-agar plates under the
Microbial Assay for Antibiotics, dispensing 5.0 mL of agar
medium for base layer and 8.0 mL of the agar medium for
seed into the Petri dish.
(vi) Standard solutions—Weigh accurately an amount of
Bleomycin A2 Hydrochloride RS, previously dried under
reduced pressure not exceeding 0.67 kPa at an ordinary tem-
perature for 3 hours, equivalent to about 15 mg (potency),
dissolve in 0.1 mol/L phosphate buffer solution (pH 6.8) to
stock solution. Keep the standard stock solution at 59C or
below, and use within 30 days. Take exactly a suitable
amount of the standard stock solution before use, add 0.1
mol/L phosphate buffer solution (pH 6.8) to make solutions
so that each mL contains 30 mg (potency) and 15 mg (po-
tency), and use these solutions as the high concentration
standard solution and low concentration standard solution,
respectively.
(vii) Sample solutions—Weigh accurately an amount of
Bleomycin Hydrochloride, equivalent to about 15 mg (po-
tency), and dissolve in 0.1 mol/L phosphate buffer solution
(pH 6.8) to make exactly 100 mL. Take exactly a suitable
amount of this solution, add 0.1 mol/L phosphate buffer so-
lution (pH 6.8) to make solutions so that each mL contains
30 mg (potency) and 15 mg (potency), and use these solutions
as the high concentration sample solution and low concen-
Bleomycinoic Acid
tration sample solution, respectively.
1-Bleomycinoic acid sulfate
Containers and storage Containers—Tight containers. Bleomycin A1
N 1-[3-(Methylsulfinyl)propyl]bleomycinamide sulfate
Bleomycin Demethyl-A2
N 1-[3-(Methylsulfanyl)propyl]bleomycinamide sulfate
Bleomycin A2
N 1-[3-(Dimethylsulfonium)propyl]bleomycinamide sulfate
Bleomycin A2?-a
N 1-(4-Aminobutyl)bleomycinamide sulfate
Bleomycin A2?-b
N 1-(3-Aminopropyl)bleomycinamide sulfate
Bleomycin A5
N 1-{3-[(4-Aminobutyl)amino]propyl}bleomycinamide
sulfate
Bleomycin B1?
Bleomycinamide sulfate
Bleomycin B2
N 1-(4-Guanidinobutyl)bleomycinamide sulfate
Bleomycin B4
N 1-{4-[3-(4-Guanidinobutyl)guanidino]butyl}-
bleomycinamide sulfate
[9041-93-4, Bleomycin Sulfate]
Bleomycin Sulfate is the sulfate of a mixture of sub-

stances having antitumor activity produced by the
growth of Streptomyces verticillus.
more than 2000 mg (potency) per mg, calculated on
the dried basis. The potency of Bleomycin Sulfate
JP XVII Official Monographs / Bleomycin Sulfate 519
is expressed as mass (potency) of bleomycin A2

(C55H84ClN17O21S3: 1451.00). Time after injection Mobile phase A Mobile phase B
of sample (min) (volz) (volz)
Description Bleomycin Sulfate occurs as a white to yellow-
ish white powder. 0 – 60 100 → 0 0 → 100
It is freely soluble in water, and slightly soluble in ethanol 60 – 75 0 100
(95).
It is hygroscopic. Flow rate: About 1.2 mL per minute.
Identification (1) To 4 mg of Bleomycin Sulfate add 5 mL Time span of measurement: Twenty minutes after elution
of copper (II) sulfate TS, and dissolve in water to make 100 of the peak of demethylbleomycin A2, beginning after the
mL. Determine the absorption spectrum of this solution as solvent peak.
directed under Ultraviolet-visible Spectrophotometry <2.24>, System suitability—
and compare the spectrum with the Reference Spectrum: System performance: When the procedure is run with 20
both spectra exhibit similar intensities of absorption at the mL of the sample solution under the above operating condi-
same wavelengths. tions, bleomycin A2 and bleomycin B2 are eluted in this order
(2) Determine the infrared absorption spectrum of with the resolution between these peaks being not less than 5.
Bleomycin Sulfate as directed in the potassium bromide disk System repeatability: When the test is repeated 6 times
method under Infrared Spectrophotometry <2.25>, and com- with 20 mL of the sample solution under the above operating
pare the spectrum with the Reference Spectrum: both spectra conditions, the relative standard deviation of the peak area
exhibit similar intensities of absorption at the same wave of bleomycin A2 is not more than 2.0z.
numbers. Purity (1) Clarity and color of solution—A solution ob-
(3) A solution of Bleomycin Sulfate (1 in 200) responds tained by dissolving 80 mg of Bleomycin Sulfate in 4 mL of
to the Qualitative Tests <1.09> (1) and (2) for sulfate. water is clear and colorless.
pH <2.54> The pH of a solution obtained by dissolving 10 (2) Copper—Dissolve exactly 75 mg of Bleomycin Sul-
mg of Bleomycin Sulfate in 20 mL of water is between 4.5 fate in 10 mL of diluted nitric acid (1 in 100), and use this so-
and 6.0. lution as the sample solution. Separately, to exactly 15 mL
of Standard Copper Solution add diluted nitric acid (1 in
Content ratio of the active principle Dissolve 10 mg of 100) to make exactly 100 mL, and use this solution as the
Bleomycin Sulfate in 20 mL of water, and use this solution standard solution. Perform the test with the sample solution
as the sample solution. Perform the test with 20 mL of the and standard solution as directed under Atomic Absorption
sample solution as directed under Liquid Chromatography Spectrophotometry <2.23> according to the following condi-
<2.01> according to the following conditions, and determine tions: the absorbance of the sample solution is not more than
each peak area by the automatic integration method, and that of the standard solution (not more than 200 ppm).
calculate their amounts by the area percentage method: the Gas: Combustible gas—Acetylene.
amount of the peak of bleomycin A2 (the first principal Supporting gas—Air.
peak) is between 55z and 70z, that of bleomycin B2 (the Lamp: Copper hollow-cathode lamp.
second principal peak) is between 25z and 32z, the total Wavelength: 324.8 nm.
amount of the peak of bleomycin A2 and bleomycin B2 is not
less than 85z, the amount of the peak of demethylbleomy- Loss on drying <2.41> Not more than 3.0z (60 mg, in
cin A2 (a peak having the relative retention time of 1.5 – 2.5 vacuum, phosphorus (V) oxide, 609 C, 3 hours. Take the
to bleomycin A2) is not more than 5.5z, and the total sample to be tested while avoiding moisture absorption).
amount of the rest peaks is not more than 9.5z. Assay Perform the test according to the Cylinder-plate
Operating conditions— method as directed under Microbial Assay for Antibiotics
Detector: An ultraviolet absorption photometer (wave- <4.02> according to the following conditions.
length: 254 nm). (i) Test organism—Mycrobacterium smegmatis ATCC
Column: A stainless steel column 4.6 mm in inside diame- 607
ter and 25 cm in length, packed with octadecylsilanized silica (ii) Agar medium for seed, base layer and transferring
gel for liquid chromatography (7 mm in particle diameter). the test organism
Column temperature: A constant temperature of about Glycerin 10.0 g
409 C. Peptone 10.0 g
Mobile phase stock solution: Dissolve 0.96 g of sodium 1- Meat extract 10.0 g
pentanesulfonate and 1.86 g of disodium dihydrogen ethyl- Sodium chloride 3.0 g
enediamine tetraacetate dihydrate in 1000 mL of water and 5 Agar 15.0 g
mL of acetic acid (100), and adjust the pH to 4.3 with am- Water 1000 mL
monia TS. Mix all the components and sterilize. Adjust the pH after
Mobile phase A: A mixture of the mobile phase stock so- sterilization to 6.9 – 7.1 with sodium hydroxide TS.
lution and methanol (9:1). (iii) Liquid media for suspending the test organism
Mobile phase B: A mixture of the mobile phase stock solu- Glycerin 10.0 g
tion and methanol (3:2). Peptone 10.0 g
Flowing of mobile phase: Control the gradient by mixing Meat extract 10.0 g
the mobile phases A and B as directed in the following table. Sodium chloride 3.0 g
Water 1000 mL
Mix all the components and sterilize. Adjust the pH after
sterilization to 6.9 – 7.1 with sodium hydroxide TS.
(iv) Preparation of seeded agar layer—Cultivate the test
organism on the slant of the agar medium for transferring
520 Boric Acid / Official Monographs JP XVII
the test organism at 279C for 40 to 48 hours, then inoculate (not more than 10 ppm).
the test organism thus obtained in 100 mL of the liquid me- (3) Arsenic <1.11>—Prepare the test solution with 0.40 g
dia for suspending the test organism, cultivate with shaking of Boric Acid according to Method 1, and perform the test
at between 259C and 279C for 5 days, and use this as the sus- (not more than 5 ppm).
pension of test organism. Store the suspension of test organ-
ism at a temperature not exceeding 59C, and use within 14
5 hours).
days. Add 0.5 mL of the suspension of test organism in 100
mL of the agar medium for seed previously kept at 489C, Assay Weigh accurately about 1.5 g of Boric Acid, previ-
mix thoroughly, and use as the seeded agar layer. ously dried, add 15 g of D-sorbitol and 50 mL of water, and
(v) Preparation of cylinder-agar plate—Proceed as di- dissolve by warming. After cooling, titrate <2.50> with 1
rected in 1.7. Preparation of cylinder-agar plates under the mol/L sodium hydroxide VS (indicator: 2 drops of phenol-
Microbial Assay for Antibiotics, dispensing 5.0 mL of agar phthalein TS).
medium for base layer and 8.0 mL of the agar medium for
seed into the Petri dish.
＝ 61.83 mg of H3BO3
(vi) Standard solutions—Weigh accurately an amount of
Bleomycin A2 Hydrochloride RS, previously dried under Containers and storage Containers—Well-closed contain-
reduced pressure not exceeding 0.67 kPa at an ordinary tem- ers.
perature for 3 hours, equivalent to about 15 mg (potency),
make exactly 100 mL, and use this solution as the standard Freeze-dried Botulism Antitoxin,
stock solution. Keep the standard stock solution at 59C or
below, and use within 30 days. Take exactly a suitable Equine
amount of the standard stock solution before use, add 0.1
乾燥ボツリヌスウマ抗毒素
mol/L phosphate buffer solution (pH 6.8) to make solutions
so that each mL contains 30 mg (potency) and 15 mg (po-
tency), and use these solutions as the high concentration Freeze-dried Botulism Antitoxin, Equine, is a prepa-
standard solution and low concentration standard solution, ration for injection which is dissolved before use.
respectively. It contains botulism antitoxin type A, botulism anti-
(vii) Sample solutions—Weigh accurately an amount of toxin type B, botulism antitoxin type E and botulism
Bleomycin Sulfate, equivalent to about 15 mg (potency), antitoxin type F in immunoglobulin of horse origin. It
dissolve in 0.1 mol/L phosphate buffer solution (pH 6.8) to may contain one, two or three of these four antitoxins.
make exactly 100 mL. Take exactly a suitable amount of this It conforms to the requirements of Freeze-dried
solution, add 0.1 mol/L phosphate buffer solution (pH 6.8) Botulism Antitoxin, Equine, in the Minimum Require-
to make solutions so that each mL contains 30 mg (potency) ments for Biological Products.
and 15 mg (potency), and use these solutions as the high con-
Description Freeze-dried Botulism Antitoxin, Equine,
centration sample solution and low concentration sample so-
becomes a colorless or yellow-brown, clear liquid or a
lution, respectively.
slightly white-turbid liquid on the addition of solvent.
Bromazepam
Boric Acid
ブロマゼパム
ホウ酸
H3BO3: 61.83
Boric Acid, when dried, contains not less than

99.5z of boric acid (H3BO3).
Description Boric Acid occurs as colorless or white, crys-
C14H10BrN3O: 316.15
tals or crystalline powder. It is odorless, and has a slight,
7-Bromo-5-(pyridin-2-yl)-1,3-dihydro-2H-1,4-
characteristic taste.
benzodiazepin-2-one
It is freely soluble in warm water, in hot ethanol (95) and
[1812-30-2]
in glycerin, soluble in water and in ethanol (95), and practi-
Bromazepam, when dried, contains not less than
The pH of a solution of 1.0 g of Boric Acid in 20 mL of
99.0z and not more than 101.0z of bromazepam
water is between 3.5 and 4.1.
(C14H10BrN3O).
Identification A solution of Boric Acid (1 in 20) responds
Description Bromazepam occurs as white to light yellowish
to the Qualitative Tests <1.09> for borate.
Purity (1) Clarity and color of solution—Dissolve 1.0 g It is freely soluble in acetic acid (100), slightly soluble in
of Boric Acid in 25 mL of water or in 10 mL of hot ethanol methanol, in ethanol (99.5) and in acetone, and practically
(95): the solution is clear and colorless. insoluble in water.
(2) Heavy metals <1.07>—Proceed with 2.0 g of Boric Melting point: about 2459 C (with decomposition).
Acid according to Method 1, and perform the test. Prepare
solution of Bromazepam in ethanol (99.5) (1 in 200,000) as
JP XVII Official Monographs / Bromhexine Hydrochloride 521
directed under Ultraviolet-visible Spectrophotometry <2.24>, Description Bromhexine Hydrochloride occurs as white,
and compare the spectrum with the Reference Spectrum: crystals or crystalline powder.
both spectra exhibit similar intensities of absorption at the It is freely soluble in formic acid, sparingly soluble in
same wavelengths. methanol, and slightly soluble in water and in ethanol (95).
(2) Determine the infrared absorption spectrum of The pH of its saturated solution is between 3.0 and 5.0.
Bromazepam, previously dried, as directed in the potassium Melting point: about 2399C (with decomposition).
Identification (1) Dissolve 3 mg of Bromhexine Hydro-
chloride in 0.01 mol/L hydrochloric acid TS to make 100
mL. Determine the absorption spectrum of the solution as
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of and compare the spectrum with the Reference Spectrum:
Bromazepam in a platinum crucible according to Method 4, both spectra exhibit similar intensities of absorption at the
and perform the test. Prepare the control solution with 2.0 same wavelengths.
mL of Standard Lead Solution (not more than 20 ppm). (2) Determine the infrared absorption spectrum of
(2) Related substances—Dissolve 50 mg of Bromazepam Bromhexine Hydrochloride, previously dried, as directed in
in 5 mL of a mixture of acetone and methanol (3:2), and use the potassium bromide disk method under Spectrophotome-
this solution as the sample solution. Pipet 1 mL of the sam- try <2.25>, and compare the spectrum with the Reference
ple solution, and add the mixture of acetone and methanol Spectrum: both spectra exhibit similar intensities of absorp-
(3:2) to make exactly 50 mL. Pipet 5 mL of this solution, tion at the same wave numbers.
add the mixture of acetone and methanol (3:2) to make ex- (3) Add 20 mL of water to 1 g of Bromhexine Hydro-
actly 50 mL, and use this solution as the standard solution. chloride. After thorough shaking, add 3 mL of sodium hy-
Perform the test with these solutions as directed under Thin- droxide TS, and extract with four 20-mL portions of diethyl
layer Chromatography <2.03>. Spot 20 mL each of the sample ether. Neutralize the water layer with dilute nitric acid: the
solution and standard solution on a plate of silica gel with solution responds to the Qualitative Tests <1.09> (2) for chlo-
fluorescent indicator for thin-layer chromatography. De- ride.
velop the plate with a mixture of ethyl acetate, ammonia so-
lution (28) and ethanol (99.5) (38:1:1) to a distance of about
Bromhexine Hydrochloride according to Method 2, and per-
12 cm, and air-dry the plate. Examine under ultraviolet light
(main wavelength: 254 nm): the spots other than the princi-
pal spot from the sample solution and the spot of the start-
ing point are not more than 2, and not more intense than the
exposure to light, using light-resistant vessels. Dissolve 50
spot from the standard solution.
mg of Bromhexine Hydrochloride in 10 mL of methanol,
C, and use this solution as the sample solution. Pipet 1 mL of
4 hours). the sample solution, and add the mobile phase to make
exactly 20 mL. Pipet 1 mL of this solution, add the mobile
phase to make exactly 25 mL, and use this solution as the
Assay Weigh accurately about 0.4 g of Bromazepam, pre- standard solution. Perform the test with exactly 5 mL each of
viously dried, dissolve in 80 mL of acetic acid (100), and the sample solution and standard solution as directed under
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- Liquid Chromatography <2.01> according to the following
metric titration). Perform a blank determination, and make conditions, and determine each peak area by the automatic
any necessary correction. integration method: each peak area other than bromhexine
obtained with the sample solution is not larger than the peak
area of bromhexine obtained with the standard solution.
＝ 31.62 mg of C14H10BrN3O
Containers and storage Containers—Well-closed contain- Detector: An ultraviolet absorption photometer (wave-
ers. length: 245 nm).
diameter and about 15 cm in length, packed with octadecyl-
Bromhexine Hydrochloride silanized silica gel for liquid chromatography (5 mm in parti-
cle diameter).
ブロムヘキシン塩酸塩 Column temperature: A constant temperature of about
409C.
phosphate in 900 mL of water, adjust the pH to 7.0 with 0.5
mol/L sodium hydroxide TS, and add water to make 1000
mL. To 200 mL of this solution add 800 mL of acetonitrile.
Flow rate: Adjust so that the retention time of bromhexine
is about 6 minutes.
C14H20Br2N2.HCl: 412.59
Selection of column: To 0.05 g of bamethane sulfate add
2-Amino-3,5-dibromo-N-cyclohexyl-N-
0.5 mL of the sample solution, and add the mobile phase to
methylbenzylamine monohydrochloride
make 10 mL. Proceed with 5 mL of this solution under the
[611-75-6]
above operating conditions, and calculate the resolution.
Use a column giving elution of bamethane and bromhexine
Bromhexine Hydrochloride, when dried, contains
not less than 98.5z of bromhexine hydrochloride
not less than 7.
(C14H20Br2N2.HCl).
522 Bromocriptine Mesilate / Official Monographs JP XVII
that the peak height of bromhexine from 5 mL of the stand- same wavelengths.
ard solution is between 5 mm and 15 mm. (3) Determine the infrared absorption spectrum of
Time span of measurement: About 2 times as long as the Bromocriptine Mesilate as directed in the paste method
retention time of bromhexine, beginning after the solvent under Infrared Spectrophotometry <2.25>, and compare the
peak. spectrum with the Reference Spectrum: both spectra exhibit
(4) Perform the test with Bromocriptine Mesilate as di-
4 hours).
rected under Flame Coloration Test <1.04> (2): a green color
Residue on ignition <2.44> Not more than 0.1z (1 g). appears.
Assay Weigh accurately about 0.5 g of Bromhexine Hydro- Optical rotation <2.49> [a]20 D : ＋95 – ＋1059[0.1 g, calcu-
chloride, previously dried, dissolve in 2 mL of formic acid, lated on the dried basis, a mixture of methanol and dichloro-
add 60 mL of acetic anhydride, and warm in a water bath at methane (1:1), 10 mL, 100 mm].
509 C for 15 minutes. After cooling, titrate <2.50> with 0.1
mol/L perchloric acid VS until the color of the solution
of Bromocriptine Mesilate in 10 mL of methanol: the solu-
changes from purple through blue-green to yellow-green
tion is clear, and has no more color than the following con-
(indicator: 2 drops of crystal violet TS). Perform a blank
trol solution.
Control solution: To 2.5 mL of Cobalt (II) Chloride CS,
Each mL of 0.1 mol/L perchloric acid VS 6.0 mL of Iron (III) Chloride CS and 1.0 mL of Copper (II)
＝ 41.26 mg of C14H20Br2N2.HCl Sulfate CS add diluted hydrochloric acid (1 in 40) to make
exactly 100 mL.
ers.
Bromocriptine Mesilate according to Method 2, and perform
Bromocriptine Mesilate exposure to light, using light-resistant vessels. Dissolve
0.10 g of Bromocriptine Mesilate in 10 mL of a mixture of
ブロモクリプチンメシル酸塩
methanol and chloroform (1:1), and use this solution as the
mixture of methanol and chloroform (1:1) to make exactly
200 mL, and use this solution as the standard solution (1).
Pipet 10 mL of the standard solution (1), add a mixture of
methanol and chloroform (1:1) to make exactly 20 mL, and
use this solution as the standard solution (2). Perform the
and standard solutions (1) and (2), as a band with 1 cm in
width, on a plate of silica gel for thin-layer chromatography.
C32H40BrN5O5.CH4O3S: 750.70
Develop the plate immediately with a mixture of dichloro-
(5?S )-2-Bromo-12?-hydroxy-2?-(1-methylethyl)-5?-
methane, 1,4-dioxane, ethanol (95) and ammonia solution
(2-methylpropyl)ergotaman-3?,6?,18-trione
(28) (1800:150:50:1) to a distance of about 10 cm, and dry
monomethanesulfonate
the plate under reduced pressure for 30 minutes. Spray
[22260-51-1]
evenly Dragendorff's TS for spraying on the plate, then
spray evenly hydrogen peroxide TS, cover the plate with a
Bromocriptine Mesilate contains not less than
glass plate, and examine: the spots other than the principal
98.0z of bromocriptine mesilate (C32H40BrN5O5.
CH4O3S), calculated on the dried basis.
spot from the standard solution (1), and the spot other than
Description Bromocriptine Mesilate occurs as a white to the principal spot, which is more intense than the spot from
pale yellowish white or pale brownish white crystalline the standard solution (2), is not more than one.
powder. It is odorless, or has a faint characteristic odor.
It is very soluble in acetic acid (100), freely soluble in
at a pressure not exceeding 0.67 kPa, 809C, 5 hours).
methanol, sparingly soluble in ethanol (95), very slightly
soluble in acetic anhydride, in dichloromethane and in chlo- Residue on ignition <2.44> Not more than 0.1z (1 g).
roform, and practically insoluble in water and in diethyl
Assay Weigh accurately about 0.6 g of Bromocriptine
ether.
Mesilate, dissolve in 80 mL of a mixture of acetic anhydride
and acetic acid (100) (7:1), and titrate <2.50> with 0.1 mol/L
Identification (1) Dissolve 2 mg of Bromocriptine Mesi- perchloric acid VS (potentiometric titration). Perform a
late in 1 mL of methanol, add 2 mL of 4-dimethylaminoben- blank determination, and make any necessary correction.
zaldehyde-ferric chloride TS, and shake: a purplish blue
color develops.
＝ 75.07 mg of C32H40BrN5O5.CH4O3S
Bromocriptine Mesilate in methanol (3 in 100,000) as di- Containers and storage Containers—Tight containers.
rected under Ultraviolet-visible Spectrophotometry <2.24>, Storage—Light-resistant, and not exceeding －189C.
JP XVII Official Monographs / Brotizolam 523
condenser and the mouth of the flask with 30 mL of water,

Bromovalerylurea and combine the washings with the solution in the conical
flask. Add 5 mL of nitric acid and exactly 30 mL of 0.1
ブロモバレリル尿素 mol/L silver nitrate VS, and titrate <2.50> the excess silver
nitrate with 0.1 mol/L ammonium thiocyanate VS (indica-
tor: 2 mL of ammonium iron (III) sulfate TS). Perform a
blank determination.
＝ 22.31 mg of C6H11BrN2O2
C6H11BrN2O2: 223.07
(2RS )-(2-Bromo-3-methylbutanoyl)urea Containers and storage Containers—Well-closed contain-
[496-67-3] ers.
Bromovalerylurea, when dried, contains not less

than 98.0z of bromovalerylurea (C6H11BrN2O2). Brotizolam
Description Bromovalerylurea occurs as colorless or white,
ブロチゾラム
crystals or crystalline powder. It is odorless, and has a
slightly bitter taste.
It is soluble in ethanol (95), sparingly soluble in diethyl
ether, and very slightly soluble in water.
It dissolves in sulfuric acid, in nitric acid and in hydro-
chloric acid, and precipitates are produced on the addition
of water.
Identification (1) Boil 0.2 g of Bromovalerylurea with C15H10BrClN4S: 393.69
5 mL of a solution of sodium hydroxide (1 in 10): the gas 2-Bromo-4-(2-chlorophenyl)-9-methyl-
evolved changes moistened red litmus paper to blue. Boil this 6H-thieno[3,2-f ][1,2,4]triazolo[4,3-a][1,4]diazepine
solution with an excess of dilute sulfuric acid: the odor of [57801-81-7]
valeric acid is perceptible.
(2) To 0.1 g of Bromovalerylurea add 0.5 g of anhydrous Brotizolam, when dried, contains not less than
sodium carbonate, and decompose thoroughly by gentle 98.5z and not more than 101.0z of brotizolam
heating. Dissolve the residue in 5 mL of hot water, cool, (C15H10BrClN4S).
acidify with acetic acid (31), and filter: the filtrate responds
Description Brotizolam occurs as a white or pale yellowish
to the Qualitative Tests <1.09> (2) for bromide.
crystalline powder.
Melting point <2.60> 151 – 1559C It is sparingly soluble in methanol, slightly soluble in
acetonitrile and in ethanol (99.5), and practically insoluble in
Purity (1) Acidity or alkalinity—To 1.5 g of
water.
Bromovalerylurea add 30 mL of water, shake for 5 minutes,
and filter: the filtrate is neutral. Identification (1) Determine the absorption spectrum of a
(2) Chloride <1.03>—Perform the test with a 10-mL por- solution of Brotizolam in methanol (1 in 100,000) as directed
tion of the filtrate obtained in (1). Prepare the control solu- under Ultraviolet-visible Spectrophotometry <2.24>, and
tion with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not compare the spectrum with the Reference Spectrum: both
more than 0.028z). spectra exhibit similar intensities of absorption at the same
(3) Sulfate <1.14>—Perform the test with 10 mL of the wavelengths.
filtrate obtained in (1). Prepare the control solution with (2) Determine the infrared absorption spectrum of
0.40 mL of 0.005 mol/L sulfuric acid VS (not more than Brotizolam as directed in the potassium bromide disk
0.038z). method under Infrared Spectrophotometry <2.25>, and com-
(4) Heavy metals <1.07>—Proceed with 2.0 g of pare the spectrum with the Reference Spectrum: both spectra
Bromovalerylurea according to Method 2, and perform the exhibit similar intensities of absorption at the same wave
test. Prepare the control solution with 2.0 mL of Standard numbers.
(5) Arsenic <1.11>—Dissolve 0.5 g of Bromovalerylurea
in 5 mL of sodium hydroxide TS, use this solution as the test Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
solution, and perform the test (not more than 4 ppm). Brotizolam according to Method 2, and perform the test.
(6) Readily carbonizable substances <1.15>—Perform the Prepare the control solution with 2.0 mL of Standard Lead
test with 0.5 g of Bromovalerylurea: the solution is not more Solution (not more than 10 ppm).
colored than Matching Fluid A. (2) Related substances—Dissolve 50 mg of Brotizolam in
50 mL of acetonitrile, and use this solution as the sample
solution. Pipet 2 mL of the sample solution, and add aceto-
2 hours).
nitrile to make exactly 100 mL. Pipet 1 mL of this solution,
Residue on ignition <2.44> Not more than 0.1z (1 g). add acetonitrile to make exactly 10 mL, and use this solution
Assay Weigh accurately about 0.4 g of Bromovalerylurea,
previously dried, in a 300-mL conical flask, add 40 mL of
sodium hydroxide TS, and boil gently for 20 minutes under a
reflux condenser. Cool, wash the lower part of the reflux
matic integration method: each peak area other than
524 Brotizolam Tablets / Official Monographs JP XVII
brotizolam from the sample solution is not larger than 1/2
times the peak area of brotizolam from the standard solu- Brotizolam Tablets
tion, and the total area of the peaks other than the peak of
brotizolam from the sample solution is not larger than the ブロチゾラム錠
peak area of brotizolam from the standard solution.
Brotizolam Tablets contain not less than 95.0z and
length: 242 nm).
brotizolam (C15H10BrClN4S: 393.69).
ter and 15 cm in length, packed with octylsilanized silica gel Method of preparation Prepare as directed under Tablets,
for liquid chromatography (5 mm in particle diameter). with Brotizolam.
Identification Shake a quantity of powdered Brotizolam
409 C.
Tablets, equivalent to 0.1 mg of Brotizolam, with 10 mL of
Mobile phase A: Dissolve 1.84 g of sodium 1-heptanesul-
methanol, and centrifuge. Determine the absorption spec-
fonate in 1000 mL of water.
trum of the supernatant liquid as directed under Ultraviolet-
Mobile phase B: Dissolve 0.46 g of sodium 1-heptanesul-
fonate in 250 mL of water and 750 mL of acetonitrile.
tween 239 nm and 243 nm.
the mobile phases A and B as directed in the following table. Purity Related substances—Use the sample solution ob-
tained in the Assay as the sample solution. Pipet 1 mL of the
Time after injection Mobile phase Mobile phase sample solution, add the mobile phase to make exactly 100
of sample (min) A (volz) B (volz) mL, and use this solution as the standard solution. Perform
0–4 63 37 standard solution as directed under Liquid Chromatography
4 – 15 63 → 12 37 → 88 <2.01> according to the following conditions, and determine
each peak area by the automatic integration method: the
total area of the peaks other than brotizolam obtained from
Flow rate: About 2 mL per minute. the sample solution is not larger than 1.5 times the peak area
Time span of measurement: About 2 times as long as the of brotizolam obtained from the standard solution.
retention time of brotizolam, beginning after the solvent Operating conditions—
peak. Detector, column, column temperature, mobile phase, and
System suitability— flow rate: Proceed as directed in the operating conditions in
Test for required detectability: Pipet 5 mL of the standard the Assay.
solution, and add acetonitrile to make exactly 20 mL. Con- Time span of measurement: About 3 times as long as the
firm that the peak area of brotizolam obtained with 5 mL of retention time of brotizolam, beginning after the solvent
this solution is equivalent to 18 to 32z of that with 5 mL of peak.
the standard solution. System suitability—
System performance: When the procedure is run with 5 mL Test for required detectability: To exactly 10 mL of the
of the standard solution under the above operating condi- standard solution add the mobile phase to make exactly 100
tions, the number of theoretical plates and the symmetry fac- mL. Confirm that the peak area of brotizolam obtained with
tor of the peak of brotizolam are not less than 5000 and not 40 mL of this solution is equivalent to 7 to 13z of that ob-
more than 2.0, respectively. tained with 40 mL of the standard solution.
System repeatability: When the test is repeated 6 times System performance: When the procedure is run with 40
with 5 mL of the standard solution under the above operating mL of the standard solution under the above operating con-
conditions, the relative standard deviation of the peak area ditions, the number of theoretical plates and the symmetry
of brotizolam is not more than 2.0z. factor of the peak of brotizolam are not less than 3000 and
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, not more than 2.0, respectively.
3 hours). System repeatability: When the test is repeated 6 times
Residue on ignition <2.44> Not more than 0.1z (1 g). ing conditions, the relative standard deviation of the peak
Assay Weigh accurately about 0.15 g of Brotizolam, previ- area of brotizolam is not more than 2.0z.
ously dried, dissolve in 75 mL of a mixture of acetic anhy- Uniformity of dosage units <6.02> Perform the test accord-
dride and acetic acid (100) (2:1), and titrate <2.50> with 0.1 ing to the following method: it meets the requirement of the
mol/L perchloric acid VS (potentiometric titration). Per- Content uniformity test.
form a blank determination in the same manner, and make To 1 tablet of Brotizolam Tablets add exactly V mL of the
any necessary correction. mobile phase so that each mL contains about 25 mg of
Each mL of 0.1 mol/L perchloric acid VS brotizolam (C15H10BrClN4S), shake for 15 minutes, centri-
＝ 19.68 mg of C15H10BrClN4S fuge, and use the supernatant liquid as the sample solution.
Then, proceed as directed in the Assay.
Storage—Light-resistant. Amount (mg) of brotizolam (C15H10BrClN4S)
＝ MS × AT/AS × V/1000
MS: Amount (mg) of brotizolam for assay taken
JP XVII Official Monographs / Bucillamine 525
in 15 minutes of Brotizolam Tablets is not less than 85z. ter and 15 cm in length, packed with octadecylsilanized silica
Start the test with 1 tablet of Brotizolam Tablets, with- gel for liquid chromatography (5 mm in particle diameter).
draw not less than 20 mL of the medium at the specified Column temperature: A constant temperature of about
minute after starting the test, and filter through a membrane 309C.
filter with a pore size not exceeding 0.5 mm. Discard the first Mobile phase: Dissolve 1.1 g of ammonium carbonate in
10 mL of the filtrate, pipet V mL of the subsequent filtrate, 1000 mL of water. To 600 mL of this solution add 500 mL of
add water to make exactly V? mL so that each mL contains acetonitrile.
about 0.14 mg of brotizolam (C15H10BrClN4S), and use this Flow rate: Adjust so that the retention time of brotizolam
solution as the sample solution. Separately, weigh accurately is about 3 minutes.
about 28 mg of brotizolam for assay, previously dried at System suitability—
1059C for 3 hours, dissolve in 10 mL of methanol, and add System performance: When the procedure is run with 40
water to make exactly 100 mL. Pipet 5 mL of this solution, mL of the standard solution under the above operating con-
add water to make exactly 200 mL. Pipet 2 mL of this solu- ditions, the number of theoretical plates and the symmetry
tion, add water to make exactly 100 mL, and use this solu- factor of the peak of brotizolam are not less than 3000 and
tion as the standard solution. Perform the test with exactly not more than 2.0, respectively.
200 mL each of the sample solution and standard solution as System repeatability: When the test is repeated 6 times
directed under Liquid Chromatography <2.01> according to with 40 mL of the standard solution under the above operat-
the following conditions, and determine the peak areas, AT ing conditions, the relative standard deviation of the peak
and AS, of brotizolam in each solution. area of brotizolam is not more than 1.0z.
Dissolution rate (z) with respect to the labeled amount Containers and storage Containers—Tight containers.
of brotizolam (C15H10BrClN4S) Storage—Light-resistant.
＝ MS × AT/AS × V?/V × 1/C × 9/20
MS: Amount (mg) of brotizolam for assay taken
C: Labeled amount (mg) of brotizolam (C15H10BrClN4S) Bucillamine
in 1 tablet
ブシラミン
Detector, column, and column temperature: Proceed as
directed in the operating conditions in the Assay.
Mobile phase: A mixture of water and acetonitrile (63:37).
Flow rate: Adjust so that the retention time of brotizolam
is about 7 minutes.
System suitability— C7H13NO3S2: 223.31
System performance: When the procedure is run with 200 (2R)-2-(2-Methyl-2-sulfanylpropanoylamino)-3-
mL of the standard solution under the above operating con- sulfanylpropanoic acid
ditions, the number of theoretical plates and the symmetry [65002-17-7]
factor of the peak of brotizolam are not less than 2000 and
not more than 2.0, respectively. Bucillamine, when dried, contains not less than
System repeatability: When the test is repeated 6 times 98.5z and not more than 101.0z of bucillamine
with 200 mL of the standard solution under the above operat- (C7H13NO3S2).
Description Bucillamine occurs as white, crystals or crys-
area of brotizolam is not more than 2.0z.
talline powder.
Assay Weigh accurately the mass of not less than 20 It is freely soluble in methanol and in ethanol (95), and
Brotizolam Tablets, and powder. Weigh accurately a portion slightly soluble in water.
of the powder, equivalent to about 0.25 mg of brotizolam
Identification (1) To 5 mL of a solution of Bucillamine (1
(C15H10BrClN4S), add exactly 10 mL of the mobile phase,
in 250) add 2 mL of sodium hydroxide TS and 2 drops of so-
and shake for 15 minutes. Centrifuge this solution, and use
dium pentacyanonitrosylferrate (III) TS: the solution reveals
the supernatant liquid as the sample solution. Separately,
a red-purple color.
weigh accurately about 25 mg of brotizolam for assay, previ-
(2) Determine the infrared absorption spectrum of Bucil-
ously dried at 1059 C for 3 hours, and dissolve in the mobile
lamine as directed in the potassium bromide disk method
phase to make exactly 50 mL. Pipet 5 mL of this solution,
actly 40 mL each of the sample solution and standard solu-
tion as directed under Liquid Chromatography <2.01> ac- Optical rotation <2.49> [a]20D : ＋33.0 – ＋36.59(after dry-
cording to the following conditions, and determine the peak ing, 2 g, ethanol (95), 50 mL, 100 mm).
areas, AT and AS, of brotizolam in each solution.
Amount (mg) of brotizolam (C15H10BrClN4S)
＝ MS × AT/AS × 1/100
Bucillamine according to Method 2, and perform the test.
MS: Amount (mg) of brotizolam for assay taken Prepare the control solution with 2.0 mL of Standard Lead
of Bucillamine according to Method 3, and perform the test
length: 240 nm).
(3) Related substances—Dissolve 60 mg of Bucillamine
526 Bucillamine Tablets / Official Monographs JP XVII
in 20 mL of a mixture of water and methanol (1:1), and use
this solution as the sample solution. Pipet 3 mL of the sam- Bucillamine Tablets
ple solution, add the mixture of water and methanol (1:1) to
make exactly 200 mL, and use this solution as the standard ブシラミン錠
solution. Immediately perform the test with exactly 20 mL
Bucillamine Tablets contain not less than 95.0z and
not more than 105.0z of the labeled amount of bucil-
lamine (C7H13NO3S2: 223.31).
tomatic integration method: the peak areas of related sub-
stances, having the relative retention time of about 2.3 and Method of preparation Prepare as directed under Tablets,
about 3.1 to bucillamine, obtained from the sample solution with Bucillamine.
are not larger than 8/15 times and 2/5 times the peak area of
Identification (1) To a quantity of powdered Bucillamine
bucillamine obtained from the standard solution, respec-
Tablets, equivalent to 0.1 g of Bucillamine, add 0.1 g of so-
tively, and the area of the peak other than the bucillamine
dium hydrogen carbonate and 10 mL of water, shake well,
and the peaks mentioned above from the sample solution is
filter, and add 1 or 2 drops of ninhydrin TS to the filtrate: it
not larger than 1/5 times the peak area of bucillamine from
exhibits a red-brown color.
the standard solution. The total area of the peaks other than
(2) To a quantity of powdered Bucillamine Tablets,
bucillamine from the sample solution is not larger than the
equivalent to 0.1 g of Bucillamine, add 25 mL of water,
peak area of bucillamine from the standard solution.
shake well, and filter. To 5 mL of the filtrate, add 2 mL of
dilute sodium hydroxide TS and 1 or 2 drops of sodium
pentacyanonitrosylferrate (III) TS: it exhibits a red-purple
length: 254 nm).
color.
ter and 15 cm in length, packed with octadecylsilanized silica Uniformity of dosage units <6.02>—Perform the Mass varia-
gel for liquid chromatography (5 mm in particle diameter). tion test, or the Content uniformity test according to the fol-
Column temperature: A constant temperature of about lowing method: it meets the requirement.
409 C. Store the sample solution and standard solution in a cold
Mobile phase: A mixture of 0.01 mol/L citric acid TS and place until performing the measurements. Take 1 tablet of
methanol (1:1). Bucillamine Tablets, add exactly 1 mL of the internal stand-
Flow rate: Adjust so that the retention time of bucillamine ard solution per 0.1 g of bucillamine (C7H13NO3S2), then add
is about 5 minutes. 3 mL of water and 6 mL of methanol per 0.1 g of bucilla-
Time span of measurement: About 7 times as long as the mine (C7H13NO3S2), and stir well until the tablet completely
retention time of bucillamine, beginning after the solvent disintegrated. To 1 mL of this solution add the mobile phase
peak. to make 25 mL, filter through a membrane filter with a pore
System suitability— size not exceeding 0.45 mm, and use the filtrate as the sample
Test for required detectability: To exactly 1 mL of the solution. Then, proceed as directed in the Assay.
standard solution add the mixture of water and methanol
Amount (mg) of bucillamine (C7H13NO3S2)
＝ MS × QT/QS × C × 1/200
bucillamine obtained with 20 mL of this solution is equiva-
lent to 7 to 13z of that obtained with 20 mL of the standard MS: Amount (mg) of bucillamine for assay taken
solution. C: Labeled amount (mg) of bucillamine (C7H13NO3S2) in 1
System performance: Dissolve 0.10 g of bucillamine and tablet
10 mg of 4-fluorobenzoic acid in 100 mL of methanol. To 10
Internal standard solution—A solution of 4-fluorobenzoic
mL of this solution add water to make exactly 50 mL. When
acid in methanol (1 in 100).
the procedure is run with 20 mL of this solution under the
above operating conditions, bucillamine and 4-fluorobenzoic Dissolution <6.10>—When the test is performed at 50 revolu-
acid are eluted in this order with the resolution between these tions per minute according to the Paddle method, using 900
peaks being not less than 3. mL of water as the dissolution medium, the dissolution rate
System repeatability: When the test is repeated 6 times in 30 minutes of Bucillamine Tablets is not less than 80z.
with 20 mL of the standard solution under the above operat- Store the sample solution and standard solution in a cold
ing conditions, the relative standard deviation of the peak place until performing the measurements. Start the test with
area of bucillamine is not more than 2.0z. 1 tablet of Bucillamine Tablets, withdraw not less than 20
mL of the medium at the specified minute after starting the
test, and filter through a membrane filter with a pore size
not exceeding 0.45 mm. Discard the first 10 mL of the fil-
Residue on ignition <2.44> Not more than 0.1z (1 g). trate, and use the subsequent filtrate as the sample solution.
Separately, weigh accurately an amount of bucillamine for
Assay Weigh accurately about 0.25 g of Bucillamine, dis-
assay equivalent to the labeled amount of the tablet, previ-
solve in 35 mL of methanol, add 15 mL of water, and titrate
ously dried in vacuum at 609C for 6 hours using phosphorus
<2.50> with 0.05 mol/L iodine VS (potentiometric titration).
(V) oxide as a dessicant, and dissolve in methanol to make
Perform a blank determination in the same manner, and
exactly 10 mL. Pipet 1 mL of this solution, add water to
Each mL of 0.05 mol/L iodine VS solution. Perform the test with exactly 20 mL each of the
＝ 11.17 mg of C7H13NO3S2 sample solution and standard solution as directed under Liq-
ditions, and determine the peak areas, AT and AS, of bucilla-
mine in each solution.
JP XVII Official Monographs / Bucumolol Hydrochloride 527
Dissolution rate (z) with respect to the labeled amount System suitability—
of bucillamine (C7H13NO3S2) System performance: When the procedure is run with 10
＝ MS × AT/AS × 1/C × 90 mL of the standard solution under the above operating con-
ditions, bucillamine and the internal standard are eluted in
MS: Amount (mg) of bucillamine for assay taken
C: Labeled amount (mg) of bucillamine (C7H13NO3S2) in 1
less than 3.
tablet
Operating conditions— with 10 mL of the standard solution under the above operat-
Detector, column, and column temperature: Proceed as ing conditions, the relative standard deviation of the ratio of
directed in the operating conditions in the Assay. the peak area of bucillamine to that of the internal standard
Mobile phase: A mixture of diluted phosphoric acid (1 in is not more than 1.0z.
Flow rate: Adjust so that the retention time of bucillamine
is about 4 minutes.
System performance: When the procedure is run with 20 Bucumolol Hydrochloride
ブクモロール塩酸塩
factor of the peak of bucillamine are not less than 3000 and
area of bucillamine is not more than 2.0z.
C17H23NO4.HCl: 341.83
Assay Store the sample solution and standard solution in a
8-{(2RS )-3-[(1,1-Dimethylethyl)amino]-2-
cold place until performing the measurements. Take 10
hydroxypropyloxy}-5-methylchromen-2-one
tablets of Bucillamine Tablets, add exactly 1 mL of the inter-
monohydrochloride
nal standard solution per 0.1 g of bucillamine (C7H13NO3S2),
[36556-75-9]
add 3 mL of water and 6 mL of methanol, and stir well until
the tablets completely disintegrated. To 1 mL of this solution
Bucumolol Hydrochloride, when dried, contains
add the mobile phase to make 25 mL, filter through a mem-
not less than 99.0z of bucumolol hydrochloride
brane filter with a pore size not exceeding 0.45 mm, and use
(C17H23NO4.HCl).
rately about 0.2 g of bucillamine for assay, previously dried Description Bucumolol Hydrochloride occurs as white,
in vacuum at 609C for 6 hours using phosphorus (V) oxide crystals or crystalline powder.
as a dessicant, add exactly 2 mL of the internal standard It is freely soluble in water, sparingly soluble in methanol
solution, and add 6 mL of water and 12 mL of methanol. To and in ethanol (95), slightly soluble in acetic acid (100), and
1 mL of this solution add the mobile phase to make 25 mL, practically insoluble in diethyl ether.
filter through a membrane filter with a pore size not Melting point: about 2289C (with decomposition).
exceeding 0.45 mm, and use this solution as the standard so-
Identification (1) Dissolve 0.01 g of Bucumolol Hydro-
lution. Perform the test with 10 mL each of the sample solu-
chloride in 10 mL of diluted ethanol (95) (1 in 2), and ob-
serve under ultraviolet light (main wavelength: 365 nm): the
solution shows a yellow-green fluorescence. Render this so-
calculate the ratios, QT and QS, of the peak area of bucilla-
lution alkaline by adding sodium hydroxide TS: the fluores-
mine to that of the internal standard.
cence disappears. Acidify the solution by adding dilute hy-
Amount (mg) of bucillamine (C7H13NO3S2) drochloric acid: the fluorescence reappears.
＝ MS × QT/QS × C × 1/200 (2) Dissolve 0.1 g of Bucumolol Hydrochloride in 5 mL
of water, and add 5 drops of Reinecke salt TS: a light red
MS: Amount (mg) of bucillamine for assay taken
C: Labeled amount (mg) of bucillamine (C7H13NO3S2) in 1
tablet
Bucumolol Hydrochloride (1 in 60,000) as directed under Ul-
Internal standard solution—A solution of 4-fluorobenzoic traviolet-visible Spectrophotometry <2.24>, and compare the
acid in methanol (1 in 100). spectrum with the Reference Spectrum: both spectra exhibit
Operating conditions— similar intensities of absorption at the same wavelengths.
Detector: An ultraviolet absorption photometer (wave- (4) Determine the infrared absorption spectrum of
length: 254 nm). Bucumolol Hydrochloride, previously dried, as directed in
Column: A stainless steel column 4.6 mm in inside diame- the potassium chloride disk method under Infrared Spectro-
ter and 15 cm in length, packed with octadecylsilanized silica photometry <2.25>, and compare the spectrum with the Ref-
gel for liquid chromatography (5 mm in particle diameter). erence Spectrum: both spectra exhibit similar intensities of
Column temperature: A constant temperature of about absorption at the same wave numbers.
409 C. (5) A solution of Bucumolol Hydrochloride (1 in 50) re-
Mobile phase: A mixture of diluted phosphoric acid (1 in sponds to the Qualitative Tests <1.09> for chloride.
Absorbance <2.24> E 11zcm (296 nm): 330 – 360 (after drying,
Flow rate: Adjust so that the retention time of bucillamine
40 mg, water, 2500 mL).
is about 5 minutes.
528 Bufetolol Hydrochloride / Official Monographs JP XVII
of Bucumolol Hydrochloride in 20 mL of water: the solution optical rotation.
is clear and colorless to pale yellow.
Identification (1) To 5 mL of a solution of Bufetolol
Hydrochloride (1 in 100) add 5 drops of Reinecke salt TS: a
Bucumolol Hydrochloride according to Method 2, and
light red precipitate is formed.
perform the test. Prepare the control solution with 2.0 mL of
Bufetolol Hydrochloride (1 in 20,000) as directed under Ul-
of Bucumolol Hydrochloride according to Method 3, and
(4) Related substances—Dissolve 0.10 g of Bucumolol
Bufetolol Hydrochloride, previously dried, as directed in the
potassium chloride disk method under Infrared Spectropho-
and add methanol to make exactly 50 mL. Pipet 5 mL of this
(4) A solution of Bufetolol Hydrochloride (1 in 50) re-
thin-layer chromatography. Develop the plate with a mixture Melting point <2.60> 153 – 1579
C
of methanol and ammonia-ammonium chloride buffer solu-
tion (pH 10.7) (30:1) to a distance of about 12 cm, and air-
of Bufetolol Hydrochloride in 10 mL of water: the solution
dry the plate. Examine under ultraviolet light (main wave-
length: 254 nm): the spots other than the principal spot from
(2) Sulfate <1.14>—Perform the test with 0.5 g of
the sample solution are not more intense than the spot from
Bufetolol Hydrochloride. Prepare the control solution with
0.40 mL of 0.005 mol/L sulfuric acid VS (not more than
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 0.038z).
5 hours). (3) Heavy metals <1.07>—Proceed with 2.0 g of
Bufetolol Hydrochloride according to Method 2, and per-
Assay Weigh accurately about 0.4 g of Bucumolol Hydro- Standard Lead Solution (not more than 10 ppm).
chloride, previously dried, add 45 mL of acetic acid (100), (4) Related substances—Dissolve 0.20 g of Bufetolol Hy-
dissolve by warming at 609 C, and cool. Add 105 mL of drochloride in 5 mL of methanol, and use this solution as the
acetic anhydride, and titrate <2.50> with 0.1 mol/L perchlo- sample solution. Pipet 1 mL of the sample solution, add
ric acid VS (potentiometric titration). Perform a blank deter- methanol to make exactly 200 mL, and use this solution as
mination, and make any necessary correction. the standard solution. Perform the test with these solutions
10 mL each of the sample solution and standard solution on a
＝ 34.18 mg of C17H23NO4.HCl
Containers and storage Containers—Well-closed contain- chromatography. Develop the plate with a mixture of chlo-
ers. roform, acetone, ethanol (95) and ammonia solution (28)
Bufetolol Hydrochloride nm): the spots other than the principal spot from the sample
ブフェトロール塩酸塩 ard solution.
4 hours).
Assay Weigh accurately about 0.4 g of Bufetolol Hydro-
C18H29NO4.HCl: 359.89 (100), add 50 mL of acetic anhydride, and titrate <2.50> with
1-(1,1-Dimethylethyl)amino-3-[2-(tetrahydrofuran- 0.1 mol/L perchloric acid VS (potentiometric titration). Per-
2-ylmethoxy)phenoxy]propan-2-ol monohydrochloride form a blank determination, and make any necessary correc-
[35108-88-4] tion.
Bufetolol Hydrochloride, when dried, contains ＝ 35.99 mg of C18H29NO4.HCl
not less than 98.5z of bufetolol hydrochloride
(C18H29NO4.HCl). Containers and storage Containers—Tight containers.
Description Bufetolol Hydrochloride occurs as white, crys-
It is freely soluble in water and in methanol, soluble in
ethanol (95) and in acetic acid (100), and practically insolu-
ble in diethyl ether.
A solution of Bufetolol Hydrochloride (1 in 10) shows no
JP XVII Official Monographs / Buformin Hydrochloride Delayed-release Tablets 529

Buformin Hydrochloride Column temperature: A constant temperature of about
359C.
ブホルミン塩酸塩 Mobile phase: A mixture of a solution of sodium
perchlorate monohydrate in diluted phosphoric acid (1 in
1000) (7 in 250) and acetonitrile (7:1).
Flow rate: Adjust so that the retention time of buformin is
about 6 minutes.
C6H15N5.HCl: 193.68 Time span of measurement: About 2 times as long as the
1-Butylbiguanide hydrochloride retention time of buformin, beginning after the solvent peak.
[1190-53-0] System suitability—
Buformin Hydrochloride, when dried, contains solution, and add the mobile phase to make exactly 10 mL.
not less than 98.5z and not more than 101.0z of Confirm that the peak area of buformin obtained from 10
buformin hydrochloride (C6H15N5.HCl). mL of this solution is equivalent to 7 to 13z of that from 10
mL of the standard solution.
Description Buformin Hydrochloride occurs as a white
crystalline powder.
Identification (1) To 5 mL of a solution of Buformin factor of the peak of buformin are not less than 5000 and
Hydrochloride (1 in 2000) add 1 mL of dilute sodium penta- not more than 2.0, respectively.
cyanonitrosylferrate (III)-potassium hexacyanoferrate (III) System repeatability: When the test is repeated 6 times
TS: a red-brown color develops. with 10 mL of the standard solution under the above operat-
(2) Determine the absorption spectrum of a solution of ing conditions, the relative standard deviation of the peak
Buformin Hydrochloride (1 in 125,000) as directed under area of buformin is not more than 1.0z.
the spectrum with the Reference Spectrum: both spectra
3 hours).
exhibit similar intensities of absorption at the same wave-
lengths. Residue on ignition <2.44> Not more than 0.1z (1 g).
Assay Weigh accurately about 0.15 g of Buformin Hydro-
Buformin Hydrochloride as directed in the potassium chlo-
chloride, previously dried, dissolve in 50 mL of a mixture of
acetic anhydride and acetic acid (100) (7:3), and immediately
same wave numbers.
(4) A solution of Buformin Hydrochloride (1 in 20) re-
sponds to the Qualitative Tests <1.09> for chlorides. Each mL of 0.1 mol/L perchloric acid VS
＝ 9.684 mg of C6H15N5.HCl
Buformin Hydrochloride according to Method 1, and per-
Standard Lead Solution (not more than 20 ppm). Buformin Hydrochloride Delayed-
(2) Arsenic <1.11>—Prepare the test solution with 1.0 g release Tablets
of Buformin Hydrochloride according to Method 1, and per-
form the test (not more than 2 ppm). ブホルミン塩酸塩腸溶錠
(3) Related substances—Dissolve 0.10 g of Buformin
Buformin Hydrochloride Delayed-release Tablets
contain not less than 93.0z and not more than
107.0z of the labeled amount of buformin hydrochlo-
ride (C6H15N5.HCl: 193.68).
solution as directed under Liquid Chromatography <2.01> Method of preparation Prepare as directed under Tablets,
according to the following conditions, and determine each with Buformin Hydrochloride.
Identification To a quantity of powdered Buformin Hy-
method: the area of the peak other than buformin obtained
drochloride Delayed-release Tablets, equivalent to 0.1 g of
from the sample solution is not larger than 1/5 times the
Buformin Hydrochloride, add 10 mL of water, shake well,
peak area of buformin obtained from the standard solution.
and then filter. To 4 mL of the filtrate add 1 mL of a mix-
Furthermore, the total of the areas of all peaks other than
ture of hydrogen peroxide TS, sodium pentacyanonitro-
the buformin peak from the sample solution is not larger
sylferrate (III) TS and a solution of sodium hydroxide (1 in
than 1/2 times the peak area of buformin from the standard
10) (2:1:1): the solution exhibits a red to red-purple color.
solution.
Operating conditions— Uniformity of dosage units <6.02> Perform the test accord-
Detector: An ultraviolet absorption photometer (wave- ing to the following method: it meets the requirement of the
length: 230 nm). Content uniformity test.
Column: A stainless steel column 4.6 mm in inside diame- To 1 tablet of Buformin Hydrochloride Delayed-release
ter and 15 cm in length, packed with octadecylsilanized silica Tablets add 5 mL of a mixture of ethanol (99.5) and acetone
530 Buformin Hydrochloride Delayed-release Tablets / Official Monographs JP XVII
(1:1), disperse the pellicle to smaller using ultrasonic waves, Flow rate: Adjust so that the retention time of buformin is
add exactly 10 mL of the internal standard solution per 50 about 6 minutes.
mg of buformin hydrochloride (C6H15N5.HCl), and then add System suitability—
diluted acetonitrile (1 in 2) to make 13V/20 mL. Disintegrate System performance: When the procedure is run with 20
the tablet using ultrasonic waves, then shake for 20 minutes, mL of the standard solution under the above operating con-
and add diluted acetonitrile (1 in 2) to make V mL so that ditions, the number of theoretical plates and the symmetry
each mL contains about 0.5 mg of buformin hydrochloride factor of the peak of buformin are not less than 3000 and
(C6H15N5.HCl) per mL. Centrifuge this solution, to 1 mL of not more than 2.0, respectively.
the supernatant liquid, add the mobile phase to make 50 mL. System repeatability: When the test is repeated 6 times
If necessary, filter this solution through a membrane filter with 20 mL of the standard solution under the above operat-
with a pore size not exceeding 0.5 mm, and use the filtrate as ing conditions, the relative standard deviation of the peak
the sample solution. Then, proceed as directed in the Assay. area of buformin is not more than 2.0z.
Amount (mg) of buformin hydrochloride (C6H15N5.HCl) Assay Add 20 mL of a mixture of ethanol (99.5) and
＝ MS × QT/QS × V/50 acetone (1:1) to an amount of Buformin Hydrochloride
Delayed-release Tablets equivalent to 0.5 g of buformin hy-
MS: Amount (mg) of buformin hydrochloride for assay
drochloride (C6H15N5.HCl), disperse the pellicles to smaller
taken
using ultrasonic waves, and then add 100 mL of diluted
Internal standard solution—A solution of p-acetanisidide in acetonitrile (1 in 2). Disintegrate the tablets with the aid of
diluted acetonitrile (1 in 2) (1 in 150). ultrasonic waves, shake for 20 minutes, and then add diluted
acetonitrile (1 in 2) to make exactly 200 mL. Centrifuge this
Dissolution <6.10> When the tests are performed at 50
solution, pipet 10 mL of the supernatant liquid, add exactly
revolutions per minute according to the Paddle method,
5 mL of the internal standard solution, and then add diluted
using 900 mL each of 1st fluid for dissolution test and 2nd
acetonitrile (1 in 2) to make 50 mL. Pipet 1 mL of this solu-
fluid for dissolution test as the dissolution medium, the dis-
tion, and add the mobile phase to make 50 mL. If necessary,
solution rate in 120 minutes of Buformin Hydrochloride
filter this solution through a membrane filter with a pore size
Delayed-release Tablets using 1st fluid is not more than 5z,
not exceeding 0.5 mm, and use the filtrate as the sample solu-
and that in 90 minutes of Buformin Hydrochloride Delayed-
tion. Separately, weigh accurately about 25 mg of buformin
release Tablets using 2nd fluid is not less than 80z.
hydrochloride for assay, previously dried at 1059C for 3
Start the test with 1 tablet of Buformin Hydrochloride
hours, dissolve in an adequate amount of diluted acetonitrile
Delayed-release Tablets, withdraw not less than 20 mL of the
(1 in 2), add exactly 5 mL of the internal standard solution,
medium at the specified minute after starting the test, and
and then add diluted acetonitrile (1 in 2) to make 50 mL. To
filter through a membrane filter with a pore size not
1 mL of this solution add the mobile phase to make 50 mL,
pipet V mL of the subsequent filtrate, add the relevant disso-
lution medium to make exactly V? mL so that each mL con-
tains about 56 mg of buformin hydrochloride (C6H15N5.
HCl), and use this solution as the sample solution. Sepa-
QT and QS, of the peak area of buformin to that of the inter-
rately, weigh accurately about 28 mg of buformin hydro-
nal standard.
chloride for assay, previously dried at 1059C for 3 hours,
and dissolve in the relevant dissolution medium to make ex- Amount (mg) of buformin hydrochloride (C6H15N5.HCl)
actly 100 mL. Pipet 4 mL of this solution, add the relevant ＝ MS × QT/QS × 20
taken
directed under Liquid Chromatography <2.01> according to Internal standard solution—A solution of p-acetanisidide in
the following conditions, and determine the peak areas, AT diluted acetonitrile (1 in 2) (1 in 150).
and AS, of buformin in each solution. Operating conditions—
length: 233 nm).
of buformin hydrochloride (C6H15N5.HCl)
＝ MS × AT/AS × V?/V × 1/C × 180
MS: Amount (mg) of buformin hydrochloride for assay gel for liquid chromatography (5 mm in particle diameter).
taken Column temperature: A constant temperature of about
C: Labeled amount (mg) of buformin hydrochloride 359C.
(C6H15N5.HCl) in 1 tablet Mobile phase: A mixture of a solution of sodium
perchlorate (7 in 250) and acetonitrile (7:1).
Flow rate: Adjust so that the retention time of buformin is
about 7 minutes.
length: 230 nm).
ditions, buformin and the internal standard are eluted in this
359 C.
than 5.
Mobile phase: A mixture of a solution of sodium
perchlorate in diluted phosphoric acid (1 in 1000) (7 in 500)
and acetonitrile (7:1).
JP XVII Official Monographs / Bumetanide 531
the peak area of buformin to that of the internal standard is the absorbances, AT and AS, at 233 nm.
not more than 1.0z.
Containers and storage Containers—Well-closed contain- of buformin hydrochloride (C6H15N5.HCl)
ers. ＝ MS × AT/AS × V?/V × 1/C × 18
taken
Buformin Hydrochloride Tablets C: Labeled amount (mg) of buformin hydrochloride
(C6H15N5.HCl) in 1 tablet
ブホルミン塩酸塩錠
Assay Weigh accurately not less than 20 Buformin Hydro-
chloride Tablets, and powder. Weigh accurately a portion of
Buformin Hydrochloride Tablets contain not less
the powder, equivalent to about 60 mg of buformin hydro-
chloride (C6H15N5.HCl), add water to make exactly 200 mL,
amount of buformin hydrochloride (C6H15N5.HCl:
and treat with ultrasonic waves for 5 minutes. Take 40 mL
193.68).
of this solution, centrifuge, pipet 2 mL of the supernatant
Method of preparation Prepare as directed under Tablets, liquid, add water to make exactly 100 mL, and use this solu-
with Buformin Hydrochloride. tion as the sample solution. Separately, weigh accurately
about 60 mg of buformin hydrochloride for assay, previ-
Identification To a quantity of powdered Buformin Hy-
ously dried at 1059C for 3 hours, and dissolve in water to
drochloride Tablets, equivalent to 1 g of Buformin Hydro-
make exactly 200 mL. Pipet 2 mL of this solution, add water
chloride, add 100 mL of water, shake well, and then filter.
To 4 mL of the filtrate add 1 mL of dilute sodium penta-
ard solution. Perform the test with the sample solution and
cyanonitrosylferrate (III)-potassium hexacyanoferrate (III)
standard solution as directed under Ultraviolet-visible Spec-
TS: the solution exhibits a red-brown color.
trophotometry <2.24>, and determine the absorbances, AT
Uniformity of dosage units <6.02> Perform the Mass varia- and AS, at 233 nm.
Amount (mg) of buformin hydrochloride (C6H15N5.HCl)
＝ M S × A T / AS
Take 1 tablet of Buformin Hydrochloride Tablets, add
water to make exactly 200 mL, and then treat with ultrasonic MS: Amount (mg) of buformin hydrochloride for assay
waves for 5 minutes. Take 40 mL of this solution and centri- taken
fuge. Pipet V mL of the supernatant liquid equivalent to
about 0.5 mg of buformin hydrochloride (C6H15N5.HCl),
ers.
mg of buformin hydrochloride for assay, previously dried at
1059C for 3 hours, and dissolve in water to make exactly 200 Bumetanide
mL. Pipet 2 mL of this solution, add water to make exactly
ブメタニド
100 mL, and use this solution as the standard solution. De-
termine the absorbances, AT and AS, of the sample solution
Amount (mg) of buformin hydrochloride (C6H15N5.HCl)
＝ MS × AT/AS × 2/V
taken
C17H20N2O5S: 364.42
Dissolution <6.10> When the test is performed at 50 revolu- 3-Butylamino-4-phenoxy-5-sulfamoylbenzoic acid
tions per minute according to the Paddle method, using 900 [28395-03-1]
in 15 minutes of Buformin Hydrochloride Tablets is not less Bumetanide, when dried, contains not less than
than 80z. 98.5z of bumetanide (C17H20N2O5S).
Start the test with 1 tablet of Buformin Hydrochloride
Description Bumetanide occurs as white, crystals or crys-
talline powder.
It is freely soluble in pyridine, soluble in methanol and in
ethanol (95), slightly soluble in diethyl ether, and practically
insoluble in water.
quent filtrate, and add water to make exactly V? mL so that
It dissolves in potassium hydroxide TS.
each mL contains about 5.6 mg of buformin hydrochloride
(C6H15N5.HCl), and use this solution as the sample solution.
Separately, weigh accurately about 28 mg of buformin hy- Identification (1) Dissolve 0.01 g of Bumetanide in 1 mL
drochloride for assay, previously dried at 1059 C for 3 hours, of pyridine, add 2 drops of copper (II) sulfate TS, shake,
and dissolve in water to make exactly 100 mL. Pipet 2 mL of add 3 mL of water and 5 mL of chloroform, shake, and
this solution, add water to make exactly 100 mL, and use allow to stand: a light blue color develops in the chloroform
this solution as the standard solution. Perform the test with layer.
the sample solution and standard solution as directed under (2) Dissolve 0.04 g of Bumetanide in 100 mL of phos-
Ultraviolet-visible Spectrophotometry <2.24>, and determine phate buffer solution (pH 7.0) and dilute 10 mL of the solu-
532 Bunazosin Hydrochloride / Official Monographs JP XVII
tion with water to make 100 mL. Determine the absorption Each mL of 0.1 mol/L sodium hydroxide VS
spectrum of the solution as directed under Ultraviolet-visible ＝ 36.44 mg of C17H20N2O5S
ties of absorption at the same wavelengths.
Bumetanide, previously dried, as directed in the potassium
bromide disk method under Infrared Spectrophotometry Bunazosin Hydrochloride
ブナゾシン塩酸塩
Purity (1) Clarity and color of solution—Dissolve 50 mg
of Bumetanide in 2 mL of a solution of potassium hydroxide
(1 in 30) and 8 mL of water: the solution is clear, and is not
more colored than the following control solution.
Control solution: Pipet 0.5 mL each of Cobalt (II) Chlo-
ride CS, Iron (III) Chloride CS and Copper (II) Sulfate CS, C19H27N5O3.HCl: 409.91
mix them, and add diluted hydrochloric acid (1 in 40) to 4-Amino-2-(4-butanoyl-1,4-diazepan-1-yl)-6,7-
make exactly 100 mL. dimethoxyquinazoline monohydrochloride
(2) Chloride <1.03>—Mix well 0.5 g of Bumetanide with [52712-76-2]
0.7 g of potassium nitrate and 1.2 g of anhydrous sodium
carbonate, transfer, in small portions, to a red-hot platinum Bunazosin Hydrochloride, when dried, contains
crucible, and heat to red-hot until the reaction is complete. not less than 98.0z of bunazosin hydrochloride
After cooling, to the residue add 14 mL of dilute sulfuric (C19H27N5O3.HCl).
acid and 6 mL of water, boil for 5 minutes, filter, wash the
Description Bunazosin Hydrochloride occurs as a white
residue with 10 mL of water, combine the filtrate and the
crystalline powder.
washing, and add 6 mL of dilute nitric acid and water to
It is very soluble in formic acid, slightly soluble in water
and in methanol, very slightly soluble in ethanol (99.5), and
solution. Prepare the control solution with 0.30 mL of 0.01
practically insoluble in diethyl ether.
Bumetanide according to Method 2, and perform the test. Identification (1) Dissolve 0.1 g of Bunazosin Hydrochlo-
Prepare the control solution with 2.0 mL of Standard Lead ride in 10 mL of 0.2 mol/L hydrochloric acid TS, and boil
Solution (not more than 10 ppm). for 3 minutes over a flame: butylic acid like odor is percepti-
(4) Arsenic <1.11>—Prepare the test solution with 1.0 g ble.
of Bumetanide according to Method 3, and perform the test (2) Determine the infrared absorption spectrum of
(not more than 2 ppm). Bunazosin Hydrochloride, previously dried, as directed in
(5) Related substances—Conduct this procedure without the potassium bromide disk method under Infrared Spectro-
exposure to light, using light-resistant vessels. Dissolve 0.10 photometry <2.25>, and compare the spectrum with the Ref-
g of Bumetanide in 10 mL of methanol, and use this solution erence Spectrum: both spectra exhibit similar intensities of
as the sample solution. Pipet 1 mL of the sample solution, absorption at the same wave numbers.
and add methanol to make exactly 100 mL. Pipet 2 mL of (3) A solution of Bunazosin Hydrochloride (1 in 100) re-
this solution, add methanol to make exactly 10 mL, and use sponds to the Qualitative Tests <1.09> for chloride.
Bunazosin Hydrochloride according to Method 4, and per-
phy <2.03>. Spot 10 mL each of the sample solution and
standard solution on a plate of silica gel with fluorescent
(2) Related substances—Dissolve 0.05 g of Bunazosin
with a mixture of chloroform, acetic acid (100), cyclohexane
and methanol (32:4:4:1) to a distance of about 12 cm, and
solution as the sample solution. To exactly 1 mL of the sam-
ple solution add the mobile phase to make exactly 200 mL,
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, <2.01> according to the following conditions. Determine each
2 hours). peak area of both solutions by the automatic integration
method: the total area of the peaks other than bunazosin
Assay Weigh accurately about 0.5 g of Bumetanide, previ- bunazosin from the standard solution.
ously dried, dissolve in 50 mL of ethanol (95), and titrate Operating conditions—
<2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric Detector: An ultraviolet absorption photometer (wave-
titration). Perform a blank determination, and make any length: 254 nm).
necessary correction. Column: A stainless steel column about 4 mm in inside
JP XVII Official Monographs / Bupivacaine Hydrochloride Hydrate 533
silanized silica gel for liquid chromatography (5 mm in parti- sodium hydroxide TS (34:15:1) shows no optical rotation.
cle diameter). Melting point: about 2529C (with decomposition).
309 C.
solution of Bupivacaine Hydrochloride Hydrate in 0.01
Mobile phase: Dissolve 1.44 g of sodium lauryl sulfate in a
mol/L hydrochloric acid TS (1 in 2000) as directed under Ul-
suitable amount of water, add 10 mL of acetic acid (100),
500 mL of acetonitrile and water to make 1000 mL.
Flow rate: Adjust so that the retention time of bunazosin
is about 5 minutes.
Selection of column: Proceed with 20 mL of a mixture of
Bupivacaine Hydrochloride Hydrate as directed in the potas-
the standard solution and a solution of procaine hydrochlo-
sium chloride disk method under Infrared Spectrophotome-
ride in the mobile phase (1 in 20,000) (1:1) under the above
operating conditions, and calculate the resolution. Use a
column giving elution of procaine and bunazosin in this
(3) A solution of Bupivacaine Hydrochloride Hydrate (1
than 3.0.
in 50) responds to the Qualitative Tests <1.09> for chloride.
that the peak height of bunazosin obtained from 20 mL of pH <2.54> The pH of a solution obtained by dissolving
the standard solution is 20 to 60z of the full-scale. 1.0 g of Bupivacaine Hydrochloride Hydrate in 100 mL of
Time span of measurement: About 6 times of the retention freshly boiled and cooled water is between 4.5 to 6.0.
time of bunazosin.
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, of Bupivacaine Hydrochloride Hydrate in 50 mL of water:
2 hours). the solution is clear and colorless.
Bupivacaine Hydrochloride Hydrate according to Method 1,
Assay Weigh accurately about 0.3 g of Bunazosin Hydro- and perform the test. Prepare the control solution with 2.0
chloride, previously dried, dissolve in 6 mL of formic acid, mL of Standard Lead Solution (not more than 20 ppm).
add exactly 15 mL of 0.1 mol/L perchloric acid, and heat for (3) 2,6-Dimethylaniline—Dissolve exactly 0.50 g of
20 minutes on a water bath. After cooling, add 20 mL of Bupivacaine Hydrochloride Hydrate in 10 mL of methanol.
acetic acid (100), and titrate <2.50> the excess perchloric acid To 2 mL of this solution add 1 mL of a freshly prepared
with 0.1 mol/L sodium acetate VS (potentiometric titration). solution of 4-dimethylaminobenzaldehyde in methanol (1 in
Perform a blank determination. 100) and 2 mL of acetic acid (100), and allow to stand for 10
minutes: the color of the solution is not more colored than
the following control solution.
＝ 40.99 mg of C19H27N5O3.HCl
Control solution: Prepare by proceeding in the same man-
Containers and storage Containers—Well-closed contain- ner as above, using 2 mL of a solution of 2,6-dimethylaniline
ers. in methanol (1 in 200,000).
Storage—Light-resistant. (4) Related substances—Dissolve 50 mg of Bupivacaine
Hydrochloride Hydrate in 2.5 mL of water, add 2.5 mL of 2
mol/L sodium hydroxide TS and 5 mL of the internal stand-
Bupivacaine Hydrochloride ard solution, shake, collect the lower layer, filter, and use
the filtrate as the sample solution. Pipet 1 mL of the sample
Hydrate solution, and add the internal standard solution to make ex-
actly 100 mL. Pipet 1 mL of this solution, add the internal
ブピバカイン塩酸塩水和物
standard solution to make exactly 10 mL, and use this solu-
under Gas Chromatography <2.02> according to the follow-
ing conditions, and determine each peak area by the auto-
matic integration method: the ratio of the area of the peak
C18H28N2O.HCl.H2O: 342.90 other than bupivacaine to the peak area of the internal
(2RS)-1-Butyl-N-(2,6-dimethylphenyl)piperidine-2-carboxamide standard obtained from the sample solution is not larger
monohydrochloride monohydrate than the ratio of the peak area of bupivacaine to that of the
[14252-80-3] internal standard obtained from the standard solution.
Internal standard solution—A solution of methyl behenate
Bupivacaine Hydrochloride Hydrate contains not in dichloromethane (1 in 20,000).
less than 98.5z and not more than 101.0z of Operating conditions—
bupivacaine hydrochloride (C18H28N2O.HCl: 324.89), Detector: A hydrogen flame-ionization detector.
calculated on the anhydrous basis. Column: A quartz tube 0.32 mm in inside diameter and
30 m in length, coated the inside surface with 5z diphenyl-
Description Bupivacaine Hydrochloride Hydrate occurs as
95z dimethylpolysiloxane for gas chromatography 0.25 mm
a white crystalline powder.
in thickness.
It is freely soluble in acetic acid (100), and soluble in
Column temperature: Rise the temperature from 1809 C to
water, in methanol and in ethanol (99.5).
2309C at the rate of 59C per minute, and maintain at 2309 C
for 5 minutes.
A solution of 0.5 g of Bupivacaine Hydrochloride Hydrate
in 50 mL of a mixture of ethanol (99.5), water and 5 mol/L
534 Bupranolol Hydrochloride / Official Monographs JP XVII
about 2509C. tube with filter paper moistened with a solution of 2,6-
Detector temperature: A constant temperature of about dibromo-N-chloro-1,4-benzoquinone monoimine in ethanol
2509C. (95) (1 in 100), and heat gently for several minutes. Expose
Carrier gas: Helium. the filter paper to ammonia gas: the filter paper acquires a
Flow rate: Adjust so that the retention time of blue color.
bupivacaine is about 10 minutes. (2) Determine the absorption spectrum of a solution of
Split ratio: 1:12. Bupranolol Hydrochloride in 0.1 mol/L hydrochloric acid
Time span of measurement: About 1.5 times as long as the TS (1 in 10,000) as directed under Ultraviolet-visible Spectro-
retention time of bupivacaine. photometry <2.24>, and compare the spectrum with the Ref-
System suitability— erence Spectrum: both spectra exhibit similar intensities of
System performance: To 1 mL of the sample solution add absorption at the same wavelengths.
the internal standard solution to make 100 mL, and use this (3) Determine the infrared absorption spectrum of
solution as the solution for system suitability test. When the Bupranolol Hydrochloride, previously dried, as directed in
procedure is run with 1 mL of the solution for system suita- the potassium chloride disk method under Infrared Spectro-
bility test under the above operating conditions, bupivacaine photometry <2.25>, and compare the spectrum with the Ref-
and the internal standard are eluted in this order with the erence Spectrum: both spectra exhibit similar intensities of
resolution between these peaks being not less than 20. absorption at the same wave numbers.
System repeatability: When the test is repeated 6 times (4) A solution of Bupranolol Hydrochloride (1 in 200)
with 1 mL of the solution for system suitability test under the responds to the Qualitative Tests <1.09> for chloride.
of the ratio of the peak area of bupivacaine to that of the in-
50 mg, 0.1 mol/L hydrochloric acid TS, 500 mL).
ternal standard is not more than 2.0z.
Melting point <2.60> 223 – 2269
C
direct titration). Purity (1) Clarity and color of solution—Dissolve 0.10 g
of Bupranolol Hydrochloride in 15 mL of water: the solu-
tion is clear and colorless.
Assay Weigh accurately about 0.5 g of Bupivacaine Hydro- (2) Acidity—Dissolve 0.10 g of Bupranolol Hydrochlo-
chloride Hydrate, dissolve in 20 mL of acetic acid (100), add ride in 15 mL of freshly boiled and cooled water, and add 1
50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L drop of methyl red TS: a light red color develops. To this so-
perchloric acid VS (potentiometric titration). Perform a lution add 0.05 mL of 0.01 mol/L sodium hydroxide VS: the
blank determination in the same manner, and make any nec- color changes to yellow.
essary correction. (3) Sulfate <1.14>—Perform the test with 0.10 g of
Bupranolol Hydrochloride. Prepare the control solution
with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more
＝ 32.49 mg of C18H28N2O.HCl
than 0.168z).
Containers and storage Containers—Tight containers. (4) Heavy metals <1.07>—Proceed with 1.0 g of
Bupranolol Hydrochloride according to Method 4, and per-
Bupranolol Hydrochloride Standard Lead Solution (not more than 20 ppm).
ブプラノロール塩酸塩 of Bupranolol Hydrochloride according to Method 3, and
(6) Related substances—Dissolve 0.30 g of Bupranolol
add methanol to make exactly 100 mL, and use this solution
C14H22ClNO2.HCl: 308.24
(2RS )-3-(2-Chloro-5-methylphenoxy)-1-(1,1-
dimethylethyl)aminopropan-2-ol monohydrochloride
tion on a plate of polyamide with fluorescent indicator for
[15148-80-8]
of methanol, ammonia solution (28) and water (16:4:1) to a
Bupranolol Hydrochloride, when dried, contains
distance of about 10 cm, and air-dry the plate. Examine
not less than 98.0z of bupranolol hydrochloride
(C14H22ClNO2.HCl).
Description Bupranolol Hydrochloride occurs as a white not more intense than the spot from the standard solution.
crystalline powder.
4 hours).
water, in ethanol (95) and in acetic acid (100), very slightly
soluble in acetic anhydride, and practically insoluble in Residue on ignition <2.44> Not more than 0.1z (1 g).
diethyl ether.
Assay Weigh accurately about 0.18 g of Bupranolol Hy-
The pH of a solution of 1.0 g of Bupranolol Hydrochlo-
of acetic anhydride and acetic acid (100) (2:1) by warming,
Identification (1) Take 0.01 g of Bupranolol Hydrochlo- cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS
ride in a test tube, mix with 25 mg of potassium iodide and (potentiometric titration). Perform a blank determination,
25 mg of oxalic acid dihydrate, cover the mouth of the test and make any necessary correction.
JP XVII Official Monographs / Buprenorphine Hydrochloride 535
Each mL of 0.1 mol/L perchloric acid VS peak area of both solutions by the automatic integration
＝ 30.82 mg of C14H22ClNO2.HCl method: the area of each peak other than buprenorphine ob-
tained from the sample solution is not larger than 1/4 times
the peak area of buprenorphine obtained from the standard
ers.
solution. Furthermore, the total area of the peaks other than
buprenorphine from the sample solution is not larger than
13/20 times the peak area of buprenorphine from the stand-
Buprenorphine Hydrochloride ard solution.
ブプレノルフィン塩酸塩
length: 288 nm).
409C.
Mobile phase: A mixture of methanol, ammonium acetate
C29H41NO4.HCl: 504.10
solution (1 in 100), and acetic acid (100) (6000:1000:1).
(2S )-2-[(5R,6R,7R,14S )-17-(Cyclopropylmethyl)-4,5-
Flow rate: Adjust so that the retention time of buprenor-
epoxy-3-hydroxy-6-methoxy-6,14-ethanomorphinan-7-yl]-
phine is about 17 minutes.
3,3-dimethylbutan-2-ol monohydrochloride
[53152-21-9]
retention time of buprenorphine, beginning after the solvent
peak.
Buprenorphine Hydrochloride, when dried, con-
tains not less than 98.5z and not more than 101.0z
of buprenorphine hydrochloride (C29H41NO4.HCl).
Description Buprenorphine Hydrochloride occurs as white Confirm that the peak area of buprenorphine obtained from
to yellowish white, crystals or a crystalline powder. 20 mL of this solution is equivalent to 7 to 13z of that ob-
It is freely soluble in methanol and in acetic acid (100), tained from 20 mL of the standard solution.
and sparingly soluble in water and in ethanol (99.5). System performance: When the procedure is run with 20
Melting point: about 2689C (with decomposition). mL of the standard solution under the above operating con-
factor of the peak of buprenorphine are not less than 6500
solution of Buprenorphine Hydrochloride (1 in 5000) as
same wavelengths.
area of buprenorphine is not more than 2.0z.
Buprenorphine Hydrochloride as directed in the potassium Loss on drying <2.41> Not more than 1.0z (1 g, 1159C,
chloride disk method under Infrared Spectrophotometry 3 hours).
the same wave numbers. Assay Weigh accurately about 0.5 g of Buprenorphine Hy-
(3) A solution of Buprenorphine Hydrochloride (1 in drochloride, previously dried, dissolve in 5 mL of acetic acid
100) responds to the Qualitative Tests <1.09> for chloride. (100), add 50 mL of acetic anhydride, and titrate <2.50> with
D : －92 – －989 (after drying,
1.0 g of Buprenorphine Hydrochloride in 200 mL of water is
Purity (1) Clarity and color of solution—A solution ob-
ers.
tained by dissolving 0.1 g of Buprenorphine Hydrochloride
in 10 mL of water is clear and colorless.
Buprenorphine Hydrochloride according to Method 4, and
(3) Related substances—Dissolve 0.10 g of Buprenor-
phine Hydrochloride in 20 mL of the mobile phase, and use
536 Busulfan / Official Monographs JP XVII
Busulfan Butenafine Hydrochloride

ブスルファンブテナフィン塩酸塩
C6H14O6S2: 246.30
Tetramethylenedimethanesulfonate
[55-98-1]
C23H27N.HCl: 353.93
N-[4-(1,1-Dimethylethyl)benzyl]-N-methyl-1-(naphthalen-
Busulfan contains not less than 98.5z of busulfan
1-yl)methylamine monohydrochloride
(C6H14O6S2), calculated on the dried basis.
[101827-46-7]
Description Busulfan occurs as a white crystalline powder.
It is slightly soluble in diethyl ether, very slightly soluble in Butenafine Hydrochloride, when dried, contains
ethanol (95), and practically insoluble in water. not less than 99.0z and not more than 101.0z of
butenafine hydrochloride (C23H27N.HCl).
Identification (1) To 0.1 g of Busulfan add 10 mL of
water and 5 mL of sodium hydroxide TS, dissolve by heat- Description Butenafine Hydrochloride occurs as white,
ing, and use this solution as the sample solution. crystals or crystalline powder.
(i) To 7 mL of the sample solution add 1 drop of potas- It is very soluble in formic acid, freely soluble in methanol
sium permanganate TS: the red-purple color of potassium and in ethanol (99.5), and slightly soluble in water.
permanganate TS changes from blue-purple through blue to The pH of a solution dissolved 0.20 g of Butenafine Hy-
green. drochloride in 100 mL of water by warming and cooled is 3.0
(ii) Acidify 7 mL of the sample solution with dilute sul- to 4.0.
furic acid, and add 1 drop of potassium permanganate TS: Melting point: about 2149 C (with decomposition).
the color of potassium permanganate TS remains.
solution of Butenafine Hydrochloride in methanol (1 in
Busulfan, previously dried, as directed in the potassium bro-
same wave numbers.
Melting point <2.60> 115 – 1189C Butenafine Hydrochloride, previously dried, as directed in
Purity (1) Sulfate <1.14>—To 1.0 g of Busulfan add 40
mL of water, and dissolve by heating. Cool in ice for 15
minutes, and filter. Wash the residue with 5 mL of water,
combine the washings with the filtrate, and add 1 mL of
(3) A solution of Butenafine Hydrochloride in dilute
dilute hydrochloric acid and water to make 50 mL. Perform
ethanol (1 in 200) responds to the Qualitative Tests <1.09> (1)
the test using this solution as the test solution. Prepare the
for chloride.
control solution with 0.40 mL of 0.005 mol/L sulfuric acid
VS (not more than 0.019z). Purity (1) Heavy metals <1.07>—Dissolve 2.0 g of
(2) Heavy metals <1.07>—Proceed with 1.0 g of Busulfan Butenafine Hydrochloride in 20 mL of ethanol (99.5), add 2
according to Method 2, and perform the test. Prepare the mL of dilute acetic acid and ethanol (99.5) to make 50 mL,
control solution with 2.0 mL of Standard Lead Solution (not and perform the test using this solution as the test solution.
more than 20 ppm). The control solution: To 2.0 mL of Standard Lead Solution
add 2 mL of dilute acetic acid, and add ethanol (99.5) to
(2) Related substances—Dissolve 30 mg of Butenafine
Residue on ignition <2.44> Not more than 0.1z (1 g). Hydrochloride in 50 mL of a mixture of water and aceto-
nitrile for liquid chromatography (3:2), and use this solution
Assay Weigh accurately about 0.2 g of Busulfan, add 40
mL of water, and boil gently under a reflux condenser for 30
add a mixture of water and acetonitrile for liquid chroma-
minutes. Cool, and titrate <2.50> with 0.1 mol/L sodium hy-
tography (3:2) to make exactly 50 mL. Pipet 1 mL of this
droxide VS (indicator: 3 drops of phenolphthalein TS).
solution, add a mixture of water and acetonitrile for liquid
Each mL of 0.1 mol/L sodium hydroxide VS chromatography (3:2) to make exactly 20 mL, and use this
＝ 12.32 mg of C6H14O6S2 solution as the standard solution. Perform the test with
ers.
method: the area of the peak, having the relative retention
time of about 0.16 to butenafine, obtained from the sample
solution is not larger than 3/10 times the peak area of
butenafine obtained from the standard solution, and the
JP XVII Official Monographs / Butenafine Hydrochloride Cream 537
area of the peak other than butenafine and the peak men-
tioned above from the sample solution is not larger than the Butenafine Hydrochloride Cream
peak area of butenafine from the standard solution.
Operating conditions— ブテナフィン塩酸塩クリーム
length: 217 nm).
Butenafine Hydrochloride Cream contains not less
amount of butenafine hydrochloride (C23H27N.HCl:
353.93).
409 C. Method of preparation Prepare as directed under Creams,
Mobile phase A: Diluted 0.5 mol/L ammonium acetate TS with Butenafine Hydrochloride.
(1 in 1000).
Identification To an amount of Butenafine Hydrochloride
Mobile phase B: Acetonitrile for liquid chromatography.
Cream, equivalent to 20 mg of Butenafine Hydrochloride,
add 20 mL of acetonitrile, and warm on a water bath to melt
the bases. Shake thoroughly, add an appropriate amount of
sodium chloride, and allow to stand for 30 minutes in an ice
Time after injection Mobile phase A Mobile phase B cold water keeping not exceeding 09C to separate out the
of sample (min) (volz) (volz) bases. Centrifuge, collect the supernatant liquid, add an ap-
propriate amount of sodium chloride to the liquid, allow to
0 – 10 60 → 20 40 → 80
stand for 1 hour in an ice cold water keeping not exceeding
10 – 60 20 80
09 C, and filter while cooling. To 1 mL of the filtrate add
methanol to make 20 mL, and determine the absorption
Flow rate: 0.4 mL per minute. spectrum of this solution as directed under Ultraviolet-
Time span of measurement: For 60 minutes after injec- visible Spectrophotometry <2.24>: it exhibits maxima be-
tion, beginning after the solvent peak. tween 272 nm and 276 nm, between 281 nm and 285 nm, be-
System suitability— tween 311 nm and 315 nm, and between 316 nm and 320 nm,
Test for required detectability: Pipet 2 mL of the standard and a shoulder between 289 nm and 299 nm.
solution, and add a mixture of water and acetonitrile for liq-
uid chromatography (3:2) to make exactly 10 mL. Confirm Assay Weigh accurately a quantity of Butenafine Hydro-
that the peak area of butenafine obtained with 10 mL of this chloride Cream, equivalent to about 5 mg of butenafine hy-
solution is equivalent to 14 to 26z of that obtained with 10 drochloride (C23H27N.HCl), add 20 mL of methanol, and
mL of the standard solution. add exactly 10 mL of the internal standard solution. Warm
System performance: When the procedure is run with 10 this in a water bath for 5 minutes, and shake vigorously for
mL of the standard solution under the above operating con- 20 minutes. Then, cool in an ice bath for 15 minutes, centri-
ditions, the number of theoretical plates and the symmetry fuge, and filter the supernatant liquid with a membrane filter
factor of the peak of butenafine are not less than 20,000 and with a pore size not exceeding 0.45 mm. Discard the first 5
0.9 to 1.2, respectively. mL of the filtrate, and use the subsequent filtrate as the
System repeatability: When the test is repeated 6 times sample solution. Separately, weigh accurately about 25 mg
with 10 mL of the standard solution under the above operat- of butenafine hydrochloride for assay, previously dried in
ing conditions, the relative standard deviation of the peak vacuum at 609 C for 3 hours using phosphorus (V) oxide as
area of butenafine is not more than 2.0z. desiccant, and dissolve in methanol to make exactly 100 mL.
Loss on drying <2.41> Not more than 0.1z (1 g, in vacu- nal standard solution, and use this solution as the standard
um, phosphorus (V) oxide, 609C, 3 hours). solution. Perform the test with 5 mL each of the sample solu-
Residue on ignition <2.44> Not more than 0.1z (1 g). tion and standard solution as directed under Liquid Chroma-
Assay Weigh accurately about 0.3 g of Butenafine Hydro- calculate the ratios, QT and QS, of the peak area of butena-
chloride, previously dried, dissolve in 5 mL of formic acid, fine to that of the internal standard.
mol/L perchloric acid VS (potentiometric titration). Per- Amount (mg) of butenafine hydrochloride (C23H27N.HCl)
form a blank determination in the same manner, and make ＝ MS × QT/QS × 1/5
any necessary correction. MS: Amount (mg) of butenafine hydrochloride for assay
Each mL of 0.1 mol/L perchloric acid VS taken
＝ 35.39 mg of C23H27N.HCl Internal standard solution—A solution of diphenyl in meth-
Containers and storage Containers—Tight containers. anol (3 in 2000).
length: 282 nm).
409C.
Mobile phase: A mixture of acetonitrile and diluted 0.5
mol/L ammonium acetate TS (1 in 500) (4:1).
538 Butenafine Hydrochloride Solution / Official Monographs JP XVII
Flow rate: Adjust so that the retention time of butenafine Column: A stainless steel column 3.0 mm in inside diame-
is about 2.5 minutes. ter and 5 cm in length, packed with octylsilanized silica gel
System suitability— for liquid chromatography (3 mm in particle diameter).
System performance: When the procedure is run with 5 mL Column temperature: A constant temperature of about
of the standard solution under the above operating condi- 409C.
tions, the internal standard and butenafine are eluted in this Mobile phase: A mixture of acetonitrile and diluted 0.5
order with the resolution between these peaks being not less mol/L ammonium acetate TS (1 in 500) (4:1).
than 6. Flow rate: Adjust so that the retention time of butenafine
System repeatability: When the test is repeated 6 times is about 2.5 minutes.
with 5 mL of the standard solution under the above operating System suitability—
conditions, the relative standard deviation of the ratio of the System performance: When the procedure is run with 5 mL
peak area of butenafine to that of the internal standard is of the standard solution under the above operating condi-
not more than 1.0z. tions, the internal standard and butenafine are eluted in this
than 6.
Butenafine Hydrochloride Solution peak area of butenafine to that of the internal standard is
not more than 1.0z.
ブテナフィン塩酸塩液
Butenafine Hydrochloride Solution is a liquid for
external use.
105.0z of the labeled amount of butenafine hydro- Butenafine Hydrochloride Spray
chloride (C23H27N.HCl: 353.93). ブテナフィン塩酸塩スプレー
Method of preparation Prepare as directed under Liquids
and Solutions for Cutaneous Application, with Butenafine
Butenafine Hydrochloride Spray contains not less
Hydrochloride.
Identification To an amount of Butenafine Hydrochloride amount of butenafine hydrochloride (C23H27N.HCl:
Solution, equivalent to 10 mg of Butenafine Hydrochloride, 353.93).
add methanol to make 200 mL, and determine the absorp-
Method of preparation Prepare as directed under Pump
Sprays for Cutaneous Application, with Butenafine Hydro-
visible Spectrophotometry <2.24>: it exhibits maxima be-
chloride.
tween 272 nm and 276 nm, between 281 nm and 285 nm, be-
tween 311 nm and 315 nm, and between 316 nm and 320 nm, Identification To an amount of Butenafine Hydrochloride
and a shoulder between 289 nm and 299 nm. Spray, equivalent to 10 mg of Butenafine Hydrochloride,
Assay To an exact volume of Butenafine Hydrochloride
Solution, equivalent to about 20 mg of butenafine hydro-
visible Spectrophotometry <2.24>: it exhibits maxima be-
chloride (C23H27N.HCl), add methanol to make exactly 50
tween 272 nm and 276 nm, between 281 nm and 285 nm,
mL. Pipet 5 mL of this solution, add exactly 4 mL of the in-
between 311 nm and 315 nm, and between 316 nm and 320
ternal standard solution, then add methanol to make 25 mL,
nm, and a shoulder between 289 nm and 299 nm.
weigh accurately about 20 mg of butenafine hydrochloride Assay To an exact volume of Butenafine Hydrochloride
for assay, previously dried in vacuum at 609C for 3 hours Spray, equivalent to about 20 mg of butenafine hydrochlo-
using phosphorus (V) oxide as desiccant, and dissolve in ride (C23H27N.HCl), add methanol to make exactly 50 mL.
methanol to make exactly 50 mL. Pipet 5 mL of this solu- Pipet 5 mL of this solution, add exactly 4 mL of the internal
tion, add exactly 4 mL of the internal standard solution, standard solution, then add methanol to make 25 mL, and
then add methanol to make 25 mL, and use this solution as use this solution as the sample solution. Separately, weigh
the standard solution. Perform the test with 5 mL each of the accurately about 20 mg of butenafine hydrochloride for
sample solution and standard solution as directed under Liq- assay, previously dried in vacuum at 609C for 3 hours using
uid Chromatography <2.01> according to the following con- phosphorus (V) oxide as desiccant, and dissolve in methanol
ditions, and calculate the ratios, QT and QS, of the peak area to make exactly 50 mL. Pipet 5 mL of this solution, add ex-
of butenafine to that of the internal standard. actly 4 mL of the internal standard solution, then add meth-
anol to make 25 mL, and use this solution as the standard
Amount (mg) of butenafine hydrochloride (C23H27N.HCl)
solution. Perform the test with 5 mL each of the sample solu-
＝ M S × QT / QS
MS: Amount (mg) of butenafine hydrochloride for assay tography <2.01> according to the following conditions, and
taken calculate the ratios, QT and QS, of the peak area of butena-
fine to that of the internal standard.
Internal standard solution—A solution of diphenyl in meth-
anol (3 in 2000). Amount (mg) of butenafine hydrochloride (C23H27N.HCl)
Operating conditions— ＝ M S × QT / QS
MS: Amount (mg) of butenafine hydrochloride for assay
length: 282 nm).
JP XVII Official Monographs / Butropium Bromide 539
taken as directed under Ultraviolet-visible Spectrophotometry

Internal standard solution—A solution of diphenyl in meth-
trum 2: both spectra exhibit similar intensities of absorption
anol (3 in 2000).
at the same wavelengths.
(3) A solution of Butropium Bromide in methanol (1 in
20) responds to the Qualitative Tests <1.09> (1) for bromide.
length: 282 nm).
Column: A stainless steel column 3.0 mm in inside diame- Optical rotation <2.49> [a]20
D : －14.0 – －17.09(after dry-
ter and 5 cm in length, packed with octylsilanized silica gel ing, 0.5 g, methanol, 20 mL, 100 mm).
Purity (1) Heavy metals <1.07>—Dissolve 1.0 g of Butro-
pium Bromide in 40 mL of ethanol (95), add 2 mL of dilute
409 C.
acetic acid and water to make 50 mL. Perform the test, using
Mobile phase: A mixture of acetonitrile and diluted 0.5
this solution as the test solution. Prepare the control solution
mol/L ammonium acetate TS (1 in 500) (4:1).
with 2.0 mL of Standard Lead Solution (not more than 20
Flow rate: Adjust so that the retention time of butenafine
ppm).
is about 2.5 minutes.
(2) Related substances—Dissolve 50 mg of Butropium
Bromide in 10 mL of the mobile phase, and use this solution
tions, the internal standard and butenafine are eluted in this
actly 5 mL each of the sample solution and standard solution
than 6.
to the following conditions. Determine each peak area of
both solutions by the automatic integration method: the
peak area, having the relative retention time about 0.5 to
peak area of butenafine to that of the internal standard is
butropium from the sample solution is not larger than 1/4
not more than 1.0z.
times the peak area from the standard solution, and the total
Containers and storage Containers—Tight containers. area of all peaks other than the peak eluted first, the peak,
Storage—Light-resistant. having the relative retention time to butropium about 0.5
and butropium peak from the sample solution is not larger
than the peak area from the standard solution.
Butropium Bromide Operating conditions—
ブトロピウム臭化物 length: 220 nm).
cle diameter).
409C.
Mobile phase: Dissolve 1.15 g of sodium lauryl sulfate in
C28H38BrNO4: 532.51
1000 mL of a mixture of acetonitrile and 0.005 mol/L sulfu-
(1R,3r,5S )-8-(4-Butoxybenzyl)-3-[(2S )-hydroxy-2-
ric acid (3:2).
phenylpropanoyloxy]-8-methyl-8-azoniabicyclo[3.2.1]octane
Flow rate: Adjust so that the retention time of butropium
bromide
is about 5 minutes.
[29025-14-7]
Selection of column: Dissolve 0.50 g of Butropium
Bromide in 9 mL of ethanol (99.5) and 1 mL of 0.1 mol/L
Butropium Bromide, when dried, contains not less
potassium hydroxide-ethanol TS, and heat at 709C for 15
than 98.0z of butropium bromide (C28H38BrNO4).
minutes. After cooling, to 1 mL of this solution add the mo-
Description Butropium Bromide occurs as white, crystals bile phase to make 100 mL. Proceed with 5 mL of this solu-
or crystalline powder. tion under the above operating conditions, and calculate the
It is very soluble in formic acid, freely soluble in metha- resolution. Use a column giving elution of the peak of butro-
nol, soluble in ethanol (95), slightly soluble in water, and pium and the peak having a ratio of the retention time about
practically insoluble in diethyl ether and in acetic anhydride. 0.7 to butropium with the resolution between these peaks
being not less than 2.5.
Identification (1) To 1 mg of Butropium Bromide add 3
drops of fuming nitric acid, and evaporate on a water bath
that the peak height of the butropium obtained from 5 mL of
to dryness. Dissolve the residue in 1 mL of N, N-dimethylfor-
the standard solution is between 10 mm and 30 mm.
mamide, and add 5 to 6 drops of tetraethylammonium hy-
Time span of measurement: About twice as long as the
droxide TS: a red-purple color develops.
retention time of butropium.
Butropium Bromide in methanol (1 in 100,000) as directed Loss on drying <2.41> Not more than 1.0z (1 g, 1059C,
under Ultraviolet-visible Spectrophotometry <2.24>, and 3 hours).
compare the spectrum with the Reference Spectrum 1: both
wavelengths. Separately, determine the absorption spectrum Assay Weigh accurately about 0.8 g of Butropium
of a solution of Butropium Bromide in methanol (1 in 5000) Bromide, previously dried, dissolve in 5 mL of formic acid,
540 Butyl Parahydroxybenzoate / Official Monographs JP XVII
add 100 mL of acetic anhydride, and titrate <2.50> with 0.1 (4) Related substances—Dissolve 50 mg of Butyl Parahy-
mol/L perchloric acid-dioxane VS (potentiometric titration). droxybenzoate in 2.5 mL of methanol, and add the mobile
Perform a blank determination, and make any necessary phase to make exactly 50 mL. Pipet 10 mL of this solution,
correction. add the mobile phase to make exactly 100 mL, and use this
solution as the sample solution. Pipet 1 mL of the sample
Each mL of 0.1 mol/L perchloric acid-dioxane VS
＝ 53.25 mg of C28H38BrNO4
Pipet 1 mL of this solution, add the mobile phase to make
Containers and storage Containers—Well-closed contain- exactly 10 mL, and use this solution as the standard solution.
ers. Perform the test with exactly 10 mL each of the sample solu-
Storage—Light-resistant. tion and standard solution as directed under Liquid Chroma-
determine each peak area by the automatic integration
Butyl Parahydroxybenzoate method: the peak area of parahydroxybenzoic acid having a
relative retention time of about 0.1 to butyl parahydroxyben-
パラオキシ安息香酸ブチル zoate obtained from the sample solution is not larger than
the peak area of butyl parahydroxybenzoate obtained from
the standard solution (0.5z). For the area of the peak of
parahydroxybenzoic acid multiply the relative response fac-
tor, 1.4. Furthermore, the area of the peak other than butyl
parahydroxybenzoate and parahydroxybenzoic acid from
the sample solution is not larger than the peak area of butyl
C11H14O3: 194.23
parahydroxybenzoate from the standard solution (0.5z),
Butyl 4-hydroxybenzoate
and the total area of the peaks other than butyl parahy-
[94-26-8]
droxybenzoate is not larger than 2 times the peak area of
This monograph is harmonized with the European Phar- butyl parahydroxybenzoate from the standard solution
macopoeia and the U.S. Pharmacopeia. The parts of the text (1.0z). For this calculation the peak area not larger than
that are not harmonized are marked with symbols ( ) 1/5 times the peak area of butyl parahydroxybenzoate from
the standard solution is excluded (0.1z).
Butyl Parahydroxybenzoate contains not less than Operating conditions—
98.0z and not more than 102.0z of butyl parahy- Detector, column, column temperature, mobile phase, and
droxybenzoate (C11H14O3). flow rate: Proceed as directed in the operating conditions in
Description the Assay.
Butyl Parahydroxybenzoate occurs as color-
less crystals or white crystalline powder.
retention time of butyl parahydroxybenzoate.
It is very soluble in methanol, freely soluble in ethanol (95)
and in acetone, and practically insoluble in water.
Identification Determine the infrared absorption spectrum suitability in the Assay.
of Butyl Parahydroxybenzoate as directed in the potassium Test for required detectability: To exactly 2 mL of the
bromide disk method under Infrared Spectrophotometry standard solution add the mobile phase to make exactly 10
<2.25>, and compare the spectrum with the Reference mL. Confirm that the peak area of butyl parahydroxybenzo-
Spectrum or the spectrum of Butyl Parahydroxybenzoate ate obtained with 10 mL of this solution is equivalent to 14 to
RS: both spectra exhibit similar intensities of absorption at 26z of that obtained with 10 mL of the standard solution.
the same wave numbers. System repeatability: When the test is repeated 6 times

Melting point <2.60> 68 – 719
C
Purity (1) Clarity and color of solution—Dissolve 1.0 g area of butyl parahydroxybenzoate is not more than 2.0z.
of Butyl Parahydroxybenzoate in ethanol (95) to make 10
mL: the solution is clear and not more intensely colored than
the following control solution. Assay Weigh accurately about 50 mg each of Butyl Parahy-
Control solution: To 5.0 mL of Cobalt (II) Chloride CS, droxybenzoate and Butyl Parahydroxybenzoate RS, dissolve
12.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) separately in 2.5 mL each of methanol, and add the mobile
Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to phase to make exactly 50 mL. Pipet 10 mL each of these so-
make 1000 mL. lutions, add the mobile phase to make exactly 100 mL, and
(2) Acidity—To 2 mL of the solution of Butyl Parahy- use these solutions as the sample solution and the standard
droxybenzoate obtained in (1) add 3 mL of ethanol (95), add solution, respectively. Perform the test with exactly 10 mL
5 mL of freshly boiled and cooled water and 0.1 mL of each of the sample solution and standard solution as directed
bromocresol green-sodium hydroxide-ethanol TS, then add under Liquid Chromatography <2.01> according to the fol-
0.1 mol/L sodium hydroxide VS until the solution shows a lowing conditions, and determine the peak areas, AT and AS,
blue color: the volume of 0.1 mol/L sodium hydroxide VS of butyl parahydroxybenzoate in each solution.
used does not exceed 0.1 mL.
(3) Amount (mg) of butyl parahydroxybenzoate (C11H14O3)
Heavy metals <1.07>—Dissolve 1.0 g of Butyl Par-
＝ M S × A T / AS
ahydroxybenzoate in 25 mL of acetone, add 2 mL of dilute
acetic acid and water to make 50 mL, and perform the test MS: Amount (mg) of Butyl Parahydroxybenzoate RS
using this solution as the test solution. Prepare the control taken
solution as follows: to 2.0 mL of Standard Lead Solution
add 25 mL of acetone, 2 mL of dilute acetic acid, and water
to make 50 mL (not more than 20 ppm).
JP XVII Official Monographs / Cadralazine 541
length: 272 nm). Melting point: about 1659C (with decomposition).

solution of Cadralazine in 0.05 mol/L sulfuric acid TS (1 in
359 C.
Mobile phase: A mixture of methanol and potassium dihy-
drogen phosphate solution (17 in 2500) (1:1).
Flow rate: 1.3 mL per minute.
Cadralazine, previously dried, as directed in the potassium
System performance: Dissolve 5 mg each of Butyl Parahy-
droxybenzoate, propyl parahydroxybenzoate and parahy-
droxybenzoic acid in the mobile phase to make exactly 100
mL. Pipet 1 mL of this solution, add the mobile phase to
make exactly 10 mL, and use this solution as the solution Purity (1) Chloride <1.03>—Dissolve 0.40 g of Cadrala-
for system suitability test (1). Separately, dissolve 5 mg of zine in 15 mL of methanol, add 6 mL of dilute nitric acid
isobutyl parahydroxybenzoate in the mobile phase to make and water to make 50 mL. Perform the test using this solu-
exactly 100 mL. Pipet 0.5 mL of this solution, add the stand- tion as the test solution. Prepare the control solution with
ard solution to make exactly 50 mL, and use this solution as 0.40 mL of 0.01 mol/L hydrochloric acid VS by adding 15
the solution for system suitability test (2). When the proce- mL of methanol, 6 mL of dilute nitric acid, and water to
dure is run with 10 mL each of the solution for system suita- make 50 mL (not more than 0.036z).
bility test (1) and (2) under the above operating conditions, (2) Heavy metals <1.07>—Proceed with 1.0 g of Cadrala-
parahydroxybenzoic acid, propyl parahydroxybenzoate, zine according to Method 4, and perform the test. Prepare
isobutyl parahydroxybenzoate and butyl parahydroxybenzo- the control solution with 2.0 mL of Standard Lead Solution
ate are eluted in this order, the relative retention times of (not more than 20 ppm).
parahydroxybenzoic acid, propyl parahydroxybenzoate and (3) Related substances—Dissolve 50 mg of Cadralazine
isobutyl parahydoxybenzoate to butyl parahydroxybenzoate in 20 mL of 0.05 mol/L sulfuric acid TS, add water to 100
are about 0.1, about 0.5 and about 0.9, respectively, the mL, and use this solution as the sample solution. Pipet 1 mL
resolution between the peaks of propyl parahydroxybenzoate of the sample solution, add water to make exactly 200 mL,
and butyl parahydroxybenzoate is not less than 5.0, and the and use this solution as the standard solution. Perform the
resolution between the peaks of isobutyl parahydroxybenzo- test with exactly 10 mL each of the sample solution and
ate and butyl parahydroxybenzoate is not less than 1.5. standard solution as directed under Liquid Chromatography
System repeatability: When the test is repeated 6 times <2.01> according to the following conditions. Determine each
with 10 mL of the standard solution under the above operat- peak area of both solutions by the automatic integration
ing conditions, the relative standard deviation of the peak method: the area of the peak, having the relative retention
area of butyl parahydroxybenzoate is not more than 0.85z. time of about 2.1 to cadralazine, obtained from the sample
Containers solution is not larger than the peak area of cadralazine ob-
and storage Containers—Well-closed contain-
tained from the standard solution, and the area of the peak
ers.
other than cadralazine and the peak mentioned above is not
larger than 2/5 times the peak area of cadralazine from the
standard solution. Furthermore, the total area of the peaks
Cadralazine other than cadralazine from the sample solution is not larger
than 2 times the peak area of cadralazine from the standard
カドララジン
solution. For the areas of the peaks, having the relative
retention time of about 0.49 and about 2.1 to cadralazine,
multiply their relative response factors, 0.65 and 1.25, re-
spectively.
length: 250 nm).
C12H21N5O3: 283.33 Column: A stainless steel column 4.6 mm in inside diame-
Ethyl 3-(6-{ethyl[(2RS)-2-hydroxypropyl]amino}pyridazin- ter and 15 cm in length, packed with octadecylsilanized silica
3-yl)carbazate gel for liquid chromatography (5 mm in particle diameter).
[64241-34-5] Column temperature: A constant temperature of about
409C.
Cadralazine, when dried, contains not less than Mobile phase: Dissolve 13.6 g of sodium acetate trihydrate
98.5z and not more than 101.0z of cadralazine in 800 mL of water, adjust the pH to 5.8 with dilute acetic
(C12H21N5O3). acid, and add water to make 1000 mL. To 860 mL of this so-
lution add 140 mL of acetonitrile.
Description Cadralazine occurs as a pale yellow to light
Flow rate: Adjust so that the retention time of cadralazine
yellow crystalline powder.
It is freely soluble in acetic acid (100), soluble in methanol,
sparingly soluble in ethanol (99.5), and slightly soluble in
retention time of cadralazine.
water.
It dissolves in 0.05 mol/L sulfuric acid TS.
A solution of Cadralazine in methanol (1 in 40) shows no
solution, and add water to make exactly 25 mL. Confirm
optical rotation.
that the peak area of cadralazine obtained from 10 mL of this
542 Cadralazine Tablets / Official Monographs JP XVII
solution is equivalent to 15 to 25z of that obtained from 10 MS: Amount (mg) of cadralazine for assay taken
in 30 minutes of Cadralazine Tablets is not less than 80z.
factor of the peak of cadralazine are not less than 4000 and
Start the test with 1 tablet of Cadralazine Tablets, with-
filter with a pore size not exceeding 0.5 mm. Discard the first
10 mL of the filtrate, pipet V mL of the subsequent filtrate,
area of cadralazine is not more than 4.0z.
Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, about 5.6 mg of cadralazine (C12H21N5O3), and use this solu-
3 hours). tion as the sample solution. Separately, weigh accurately
about 30 mg of cadralazine for assay, previously dried at
1059C for 3 hours, and dissolve in water to make exactly 200
Assay Weigh accurately about 0.5 g of Cadralazine, previ- mL. Pipet 4 mL of this solution, add water to make exactly
ously dried, dissolve in 50 mL of acetic acid (100), and titrate 100 mL, and use this solution as the standard solution. De-
<2.50> with 0.1 mol/L perchloric acid VS (potentiometric termine the absorbances, AT and AS, of the sample solution
titration). Perform a blank determination in the same man- and standard solution at 254 nm as directed under Ultravio-
ner, and make any necessary correction. let-visible Spectrophotometry <2.24>.
Each mL of 0.1 mol/L perchloric acid VS Dissolution rate (z) with respect to the labeled amount
＝ 28.33 mg of C12H21N5O3 of cadralazine (C12H21N5O3)
＝ MS × AT/AS × V?/V × 1/C × 18
ers. MS: Amount (mg) of cadralazine for assay taken
C: Labeled amount (mg) of cadralazine (C12H21N5O3) in 1
tablet
Cadralazine Tablets Assay To 10 Cadralazine Tablets add 70 mL of 0.05 mol/L
sulfuric acid TS, shake well to disintegrate, add 0.05 mol/L
カドララジン錠
sulfuric acid TS to make exactly 200 mL. Centrifuge this so-
lution, pipet a volume of the supernatant liquid, equivalent
Cadralazine Tablets contain not less than 95.0z to about 2.5 mg of cadralazine (C12H21N5O3), add exactly 5
and not more than 105.0z of the labeled amount of mL of the internal standard solution, add water to make 25
cadralazine (C12H21N5O3: 283.33). mL, and use this solution as the sample solution. Separately,
weigh accurately about 25 mg of cadralazine for assay,
previously dried at 1059C for 3 hours, and dissolve in 0.05
with Cadralazine.
mol/L of sulfuric acid TS to make exactly 50 mL. Pipet 5
Identification To a quantity of powdered Cadralazine mL of this solution, add exactly 5 mL of the internal stand-
Tablets, equivalent to 20 mg of Cadralazine, add 50 mL of ard solution, add water to make 25 mL, and use this solution
0.05 mol/L sulfuric acid TS, shake well, and centrifuge. To as the standard solution. Perform the test with 5 mL each of
1 mL of the supernatant liquid add 0.05 mol/L sulfuric acid the sample solution and standard solution as directed under
TS to make 50 mL. Determine the absorption spectrum of Liquid Chromatography <2.01> according to the following
this solution as directed under Ultraviolet-visible Spectro- conditions, and calculate the ratios, QT and QS, of the peak
photometry <2.24>: it exhibits a maximum between 247 nm area of cadralazine to that of the internal standard.
and 251 nm.
Amount (mg) of cadralazine (C12H21N5O3)
Uniformity of dosage units <6.02> Perform the test accord- ＝ MS × QT/QS × 1/10
MS: Amount (mg) of cadralazine for assay taken
To 1 tablet of Cadralazine Tablets add 30 mL of 0.05 Internal standard solution—A solution of p-toluenesul-
mol/L sulfuric acid TS, shake well to disintegrate, and add fonamide in acetonitrile (1 in 50).
0.05 mol/L sulfuric acid TS to make exactly 50 mL. Centri- Operating conditions—
fuge this solution, pipet 3 mL of the supernatant liquid, add Detector: An ultraviolet absorption photometer (wave-
0.05 mol/L sulfuric acid TS to make exactly V mL so that length: 250 nm).
each mL contains about 6 mg of cadralazine (C12H21N5O3), Column: A stainless steel column 4.6 mm in inside diame-
and use this solution as the sample solution. Separately, ter and 15 cm in length, packed with octadecylsilanized silica
weigh accurately about 20 mg of cadralazine for assay, gel for liquid chromatography (5 mm in particle diameter).
previously dried at 1059C for 3 hours, and dissolve in 0.05 Column temperature: A constant temperature of about
mol/L sulfuric acid TS to make exactly 100 mL. Pipet 3 mL 409C.
of this solution, add 0.05 mol/L sulfuric acid TS to make Mobile phase: Dissolve 13.6 g of sodium acetate trihydrate
exactly 100 mL, and use this solution as the standard solu- in 800 mL of water, adjust the pH to 5.8 with dilute acetic
tion. Determine the absorbances, AT and AS, of the sample acid, and add water to make 1000 mL. To 860 mL of this so-
solution and standard solution at 249 nm as directed under lution add 140 mL of acetonitrile.
Ultraviolet-visible Spectrophotometry <2.24>. Flow rate: Adjust so that the retention time of cadralazine
Amount (mg) of cadralazine (C12H21N5O3)
＝ MS × AT/AS × V/200
JP XVII Official Monographs / Caffeine Hydrate 543
System suitability— (2) Sulfate <1.14>—To 40 mL of the sample solution ob-

System performance: When the procedure is run with 5 mL tained in (1) add 1 mL of dilute hydrochloric acid and water
of the standard solution under the above operating condi- to make 50 mL. Perform the test using this solution as the
tions, cadralazine and the internal standard are eluted in this test solution. Prepare the control solution with 0.40 mL of
order with the resolution between these peaks being not less 0.005 mol/L sulfuric acid VS (not more than 0.024z).
than 3. (3) Heavy metals <1.07>—Proceed with 2.0 g of Anhy-
System repeatability: When the test is repeated 6 times drous Caffeine according to Method 2, and perform the test.
with 5 mL of the standard solution under the above operating Prepare the control solution with 2.0 mL of Standard Lead
conditions, the relative standard deviation of the ratio of the Solution (not more than 10 ppm).
peak area of cadralazine to that of the internal standard is (4) Related substances—Dissolve 0.10 g of Anhydrous
not more than 1.0z. Caffeine in 10 mL of chloroform, and use this solution as
add chloroform to make exactly 100 mL. Pipet 1 mL of this
ers.
solution, add chloroform to make exactly 10 mL, and use
Anhydrous Caffeine phy <2.03>. Spot 10 mL each of the sample solution and
無水カフェイン
with a mixture of chloroform and ethanol (95) (9:1) to a dis-
ultraviolet light (main wavelength: 254 nm): the spots other
than the principal spot from the sample solution are not
C8H10N4O2: 194.19 test using 0.5 g of Anhydrous Caffeine: the solution is not
1,3,7-Trimethyl-1H-purine-2,6(3H,7H )-dione more colored than Matching Fluid D.
[58-08-2]
4 hours).
Anhydrous Caffeine, when dried, contains not less
than 98.5z of caffeine (C8H10N4O2). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Description Anhydrous Caffeine occurs as white, crystals Assay Weigh accurately about 0.4 g of Anhydrous
or powder. It is odorless, and has a bitter taste. Caffeine, previously dried, dissolve in 70 mL of a mixture of
It is freely soluble in chloroform, sparingly soluble in acetic anhydride and acetic acid (100) (6:1), and titrate <2.50>
water, in acetic acid (100) and in acetic anhydride, and with 0.1 mol/L perchloric acid VS until the solution changes
slightly soluble in ethanol (95) and in diethyl ether. from purple through green to yellow (indicator: 3 drops of
The pH of a solution of 1.0 g of Anhydrous Caffeine in crystal violet TS). Perform a blank determination, and make
100 mL of water is between 5.5 and 6.5. any necessary correction.
Identification (1) To 2 mL of a solution of Anhydrous Each mL of 0.1 mol/L perchloric acid VS
Caffeine (1 in 500) add tannic acid TS dropwise: a white ＝ 19.42 mg of C8H10N4O2
precipitate, which dissolves upon the dropwise addition of
tannic acid TS, is produced.
(2) To 0.01 g of Anhydrous Caffeine add 10 drops of
hydrogen peroxide TS and 1 drop of hydrochloric acid, and
evaporate on a water bath to dryness: the residue acquires a Caffeine Hydrate
yellow-red color. Invert the residue over a vessel containing
カフェイン水和物
2 to 3 drops of ammonia TS: the color turns red-purple, and
disappears upon the addition of 2 to 3 drops of sodium hy-
droxide TS.
(3) Dissolve 0.01 g of Anhydrous Caffeine in water to
make 50 mL. To 5 mL of this solution add 3 mL of diluted
acetic acid (31) (3 in 100) and 5 mL of pyridine (1 in 10),
mix, add 2 mL of diluted sodium hypochlorite TS (1 in 5),
and allow to stand for 1 minute. Add 2 mL of sodium thio- C8H10N4O2.H2O: 212.21
sulfate TS and 5 mL of sodium hydroxide TS to the solution: 1,3,7-Trimethyl-1H-purine-2,6(3H,7H )-dione
a yellow color develops. monohydrate
[5743-12-4]
Purity (1) Chloride <1.03>—Dissolve 2.0 g of Anhydrous Caffeine Hydrate, when dried, contains not less
Caffeine in 80 mL of hot water, cool rapidly to 209C, add than 98.5z of caffeine (C8H10N4O2: 194.19).
Description Caffeine Hydrate occurs as white, soft crystals
solution. To 40 mL of the sample solution add 6 mL of
or powder. It is odorless, and has a slightly bitter taste.
dilute nitric acid and water to make 50 mL. Perform the test
It is freely soluble in chloroform, sparingly soluble in
using this solution as the test solution. Prepare the control
water, in acetic acid (100) and in acetic anhydride, slightly
solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS
soluble in ethanol (95), and very slightly soluble in diethyl
544 Caffeine and Sodium Benzoate / Official Monographs JP XVII
ether. purple through green to yellow (indicator: 3 drops of crystal
The pH of a solution of 1.0 g of Caffeine Hydrate in 100 violet TS). Perform a blank determination, and make any
mL of water is between 5.5 and 6.5. necessary correction.
It effloresces in dry air.
Identification (1) To 2 mL of a solution of Caffeine Hy- ＝ 19.42 mg of C8H10N4O2
drate (1 in 500) add tannic acid TS dropwise: a white precipi-
tate, which dissolves upon the dropwise addition of tannic
acid TS, is produced.
(2) To 0.01 g of Caffeine Hydrate add 10 drops of
hydrogen peroxide TS and 1 drop of hydrochloric acid, and Caffeine and Sodium Benzoate
evaporate to dryness on a water bath: the residue acquires a
安息香酸ナトリウムカフェイン
yellow-red color. Invert the residue over a vessel containing
2 to 3 drops of ammonia TS: the color turns red-purple, and
disappears upon the addition of 2 to 3 drops of sodium hy- Caffeine and Sodium Benzoate, when dried, con-
droxide TS. tains not less than 48.0z and not more than 50.0z of
(3) Dissolve 0.01 g of Caffeine Hydrate in water to make caffeine (C8H10N4O2: 194.19), and not less than 50.0z
50 mL. To 5 mL of this solution add 3 mL of diluted acetic and not more than 52.0z of sodium benzoate
acid (31) (3 in 100) and 5 mL of a solution of pyridine (1 in (C7H5NaO2: 144.10).
10), mix, add 2 mL of diluted sodium hypochlorite TS (1 in
Description Caffeine and Sodium Benzoate occurs as a
5), and allow to stand for 1 minute. Add 2 mL of sodium
white powder. It is odorless, and has a slightly bitter taste.
thiosulfate TS and 5 mL of sodium hydroxide TS to the solu-
It is freely soluble in water, soluble in acetic acid (100) and
tion: a yellow color develops.
in acetic anhydride, sparingly soluble in ethanol (95), and
Melting point <2.60> 235 – 2389C (after drying). practically insoluble in diethyl ether.
Purity (1) Chloride <1.03>—Dissolve 2.0 g of Caffeine Identification (1) Dissolve 1 g of Caffeine and Sodium
Hydrate in 80 mL of hot water, cool rapidly to 209C, add Benzoate in 10 mL of water in a separator, add 1 drop of
water to make 100 mL, and use this solution as the sample phenolphthalein TS, and add carefully 0.01 mol/L sodium
solution. To 40 mL of the sample solution add 6 mL of hydroxide VS dropwise until a faint red color develops. Ex-
dilute nitric acid and water to make 50 mL. Perform the test tract with three 20-mL portions of chloroform by thorough
using this solution as the test solution. Prepare the control shaking, and separate the chloroform layer from the water
solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS layer. [Use the water layer for test (2).] Filter the combined
(not more than 0.011z). chloroform extracts, evaporate the filtrate to dryness on a
(2) Sulfate <1.14>—To 40 mL of the sample solution ob- water bath, and proceed the following tests with the residue:
tained in (1) add 1 mL of dilute hydrochloric acid and water (i) To 2 mL of a solution of the residue (1 in 500) add
to make 50 mL. Perform the test using this solution as the tannic acid TS dropwise: a white precipitate, which dissolves
test solution. Prepare the control solution with 0.40 mL of upon the dropwise addition of tannic acid TS, is produced.
0.005 mol/L sulfuric acid VS (not more than 0.024z). (ii) To 0.01 g of the residue add 10 drops of hydrogen
(3) Heavy metals <1.07>—Proceed with 2.0 g of Caffeine peroxide TS and 1 drop of hydrochloric acid, evaporate to
Hydrate according to Method 2, and perform the test. Pre- dryness on a water bath: the residue acquires a yellow-red
pare the control solution with 2.0 mL of Standard Lead So- color. Invert the residue over a vessel containing 2 to 3 drops
lution (not more than 10 ppm). of ammonia TS: the color turns red-purple, and disappears
(4) Related substances—Dissolve 0.10 g of Caffeine Hy- upon the addition of 2 to 3 drops of sodium hydroxide TS.
drate in 10 mL of chloroform, and use this solution as the (iii) Dissolve 0.01 g of the residue in water to make 50
sample solution. Pipet 1 mL of the sample solution, and add mL. To 5 mL of this solution add 3 mL of diluted acetic acid
chloroform to make exactly 100 mL. Pipet 1 mL of this solu- (31) (3 in 100) and 5 mL of a solution of pyridine (1 in 10),
tion, add chloroform to make exactly 10 mL, and use this mix, add 2 mL of diluted sodium hypochlorite TS (1 in 5),
solution as the standard solution. Perform the test as di- and allow to stand for 1 minute. Add 2 mL of sodium thio-
rected under Thin-layer Chromatography <2.03>. Spot 10 mL sulfate TS and 5 mL of sodium hydroxide TS to the solution:
each of the sample solution and standard solution on a plate a yellow color develops.
of silica gel with fluorescent indicator for thin-layer chroma- (2) To 5 mL of the water layer obtained in (1) add 5 mL
tography. Develop the plate with a mixture of chloroform of water: the solution responds to the Qualitative Tests
and ethanol (95) (9:1) to a distance of about 10 cm, and air- <1.09> (2) for benzoate.
dry the plate. Examine under ultraviolet light (main wave- (3) Heat Caffeine and Sodium Benzoate: white fumes
length: 254 nm): the spots other than the principal spot from are evolved. Ignite furthermore, and to the residue add hy-
the sample solution are not more intense than the spot from drochloric acid: bubbles are produced, and the solution re-
the standard solution. sponds to the Qualitative Tests <1.09> (1) for sodium salt.
test using 0.5 g of Caffeine Hydrate: the solution is not more
of Caffeine and Sodium Benzoate in 5 mL of water: the so-
colored than Matching Fluid D.
lution is clear and colorless.
Loss on drying <2.41> 0.5 – 8.5z (1 g, 809C, 4 hours). (2) Alkalinity—Dissolve 1.0 g of Caffeine and Sodium
Benzoate in 20 mL of water, and add 1 or 2 drops of phenol-
phthalein TS: no red color develops.
Assay Weigh accurately about 0.4 g of Caffeine Hydrate, (3) Chloride <1.03>—Dissolve 0.5 g of Caffeine and So-
previously dried, dissolve in 70 mL of a mixture of acetic an- dium Benzoate in 10 mL of water, and add 30 mL of ethanol
hydride and acetic acid (100) (6:1), and titrate <2.50> with 0.1 (95), 6 mL of dilute nitric acid and water to make 50 mL.
mol/L perchloric acid VS until the solution changes from Perform the test using this solution as the test solution. Pre-
JP XVII Official Monographs / Calcitonin Salmon 545
pare the control solution with 0.70 mL of 0.01 mol/L hydro- lence point to the second equivalence point (potentiometric
chloric acid VS, 30 mL of ethanol (95) and water to make 50 titration).
mL (not more than 0.050z).
(4) Chlorinated compounds—Dissolve 1.0 g of Caffeine
＝ 19.42 mg of C8H10N4O2
and Sodium Benzoate in 40 mL of water, add 10 mL of
dilute sulfuric acid, and extract with two 20-mL portions of Containers and storage Containers—Well-closed contain-
diethyl ether. Allow the combined diethyl ether extracts to ers.
evaporate at room temperature to dryness. Place this residue
and 0.7 g of calcium carbonate in a crucible, mix with a
small amount of water, and dry. Ignite at about 6009C, dis- Calcitonin Salmon
solve the residue in 20 mL of dilute nitric acid, and filter.
Wash the residue with 15 mL of water, combine the filtrate カルシトニンサケ
and the washings, and add water to make 50 mL. To this so-
lution add 0.5 mL of silver nitrate TS: the solution is not
more turbid than the following control solution to which 0.5
mL of silver nitrate TS has been added. C145H240N44O48S2: 3431.85
Control solution: Dissolve 0.7 g of calcium carbonate in [47931-85-1]
20 mL of dilute nitric acid, and filter. Wash the residue with
15 mL of water, combine the filtrate and the washings, and Calcitonin Salmon is a synthetic polypeptide con-
add 1.2 mL of 0.01 mol/L hydrochloric acid VS and water sisting of 32 amino acid residues. It is a hormone with
to make 50 mL. a blood calcium lowering effect.
(5) Heavy metals <1.07>—Dissolve 2.0 g of Caffeine and It contains not less than 4000 Units of calcitonin
Sodium Benzoate in 47 mL of water, add slowly, with salmon per 1 mg of peptide.
vigorous stirring, 3 mL of dilute hydrochloric acid, and
Description Calcitonin Salmon occurs as a white powder.
filter. Discard the first 5 mL of the filtrate, neutralize the
It is freely soluble in water.
subsequent 25 mL of the filtrate with ammonia TS, and add
It dissolves in dilute acetic acid.
2 mL of dilute acetic acid and water to make 50 mL. Per-
Dissolve 20 mg of Calcitonin Salmon in 2 mL of water:
the pH of the solution is between 5.0 and 7.0.
It is hygroscopic.
by adding 2 mL of dilute acetic acid and water to make 50
mL (not more than 20 ppm). Identification Dissolve 1 mg of Calcitonin Salmon in 1 mL
(6) Arsenic <1.11>—Prepare the test solution with 1.0 g of dilute acetic acid. Determine the absorption spectrum of
of Caffeine and Sodium Benzoate according to Method 1, this solution as directed under Ultraviolet-visible Spectro-
and perform the test (not more than 2 ppm). photometry <2.24>, and compare the spectrum with the Ref-
(7) Phthalic acid—To 0.10 g of Caffeine and Sodium erence Spectrum: both spectra exhibit similar intensities of
Benzoate add 1 mL of water and 1 mL of resorcinol-sulfuric absorption at the same wavelengths.
acid TS, and heat the mixture in an oil bath heated at a tem-
Absorbance <2.24> E 11zcm (275 nm): 3.3 – 4.0 (1 mg, dilute
perature between 1209C and 1259C to evaporate the water,
acetic acid, 1 mL).
then heat the residue for further 90 minutes, cool, and dis-
solve in 5 mL of water. To 1 mL of the solution add 10 mL Optical rotation <2.49> [a]20 D : －24 – －329(25 mg, diluted
of a solution of sodium hydroxide (43 in 500), shake, then acetic acid (100) (1 in 2), 10 mL, 100 mm).
examine under light at a wavelength between 470 nm and 490
Constituent amino acids Weigh accurately about 1 mg of
nm: the green fluorescence of the solution is not more in-
Calcitonin Salmon, put in a test tube for hydrolysis, dissolve
tense than that of the following control solution.
in 0.5 mL of diluted hydrochloric acid (1 in 2), freeze in a
Control solution: Dissolve 61 mg of potassium hydrogen
dry ice-acetone bath, seal the tube under reduced pressure,
phthalate in water to make exactly 1000 mL. Pipet exactly 1
and heat at 110 ± 29C for 24 hours. After cooling, open the
mL of the solution, add 1 mL of resorcinol-sulfuric acid TS,
tube, evaporate the hydrolyzate to dryness under reduced
and proceed as directed above.
pressure, dissolve the residue in exactly 5 mL of 0.02 mol/L
(8) Readily carbonizable substances <1.15>—Proceed
hydrochloric acid TS, and use this solution as the sample
with 0.5 g of Caffeine and Sodium Benzoate, and perform
solution. Separately, weigh accurately about 27 mg of L-
the test: the solution is not more colored than Matching
aspartic acid, about 24 mg of L-threonine, about 21 mg of L-
Fluid A.
serine, about 29 mg of L-glutamic acid, about 23 mg of L-
Loss on drying <2.41> Not more than 3.0z (2 g, 809C, proline, about 15 mg of glycine, about 18 mg of L-alanine,
4 hours). about 23 mg of L-valine, about 48 mg of L-cystine, about 30
mg of methionine, about 26 mg of L-isoleucine, about 26 mg
Assay (1) Sodium benzoate—Weigh accurately about
of L-leucine, about 36 mg of L-tyrosine, about 33 mg of
0.2 g of Caffeine and Sodium Benzoate, previously dried,
phenylalanine, about 37 mg of L-lysine hydrochloride, about
dissolve by warming in 50 mL of a mixture of acetic anhy-
42 mg of L-histidine hydrochloride monohydrate and about
dride and acetic acid (100) (6:1), cool, and titrate <2.50> with
42 mg of L-arginine hydrochloride, dissolve them in 10 mL
0.1 mol/L perchloric acid-dioxane VS to the first equiva-
of 1 mol/L hydrochloric acid TS, and add water to make
lence point (potentiometric titration). Perform a blank deter-
exactly 100 mL. Pipet 5 mL of this solution, add 0.02 mol/L
mination, and make any nccessary correction.
hydrochloric acid TS to make exactly 50 mL, and use this so-
Each mL of 0.1 mol/L perchloric acid-dioxane VS lution as the standard solution. Perform the test with exactly
＝ 14.41 mg of C7H5NaO2 10 mL each of the sample solution and standard solution as
(2) Caffeine—Continue the titration <2.50> in (1) with
the following conditions: 13 peaks of amino acids appear on
0.1 mol/L perchloric acid-dioxane VS from the first equiva-
546 Calcitonin Salmon / Official Monographs JP XVII
the chromatogram obtained from the sample solution, and System suitability—
their respective molar ratios with respect to leucine (＝5) are System performance: When the procedure is run with 10
1.9 – 2.3 for lysine, 0.8 – 1.1 for histidine, 0.9 – 1.1 for argi- mL of the standard solution under the above operating con-
nine, 1.9 – 2.1 for aspartic acid, 4.5 – 4.9 for threonine, 3.2 – ditions, aspartic acid, threonine, serine, glutamic acid, pro-
3.8 for serine, 2.8 – 3.1 for glutamic acid, 1.9 – 2.4 for proline, glycine, alanine, cystine, valine, methionine, isoleucine,
line, 2.7 – 3.3 for glycine, 1.5 – 2.5 for 1/2 cystine, 0.9 – 1.0 leucine, tyrosine, phenylalanine, lysine, histidine and argi-
for valine, and 0.8 – 1.0 for tyrosine. nine are eluted in this order with the resolutions between the
Operating conditions— peaks of threonine and serine, glycine and alanine, and
Detector: A visible spectrophotometer (wavelength: 440 isoleucine and leucine being not less than 1.2, 1.0 and 1.2,
nm and 570 nm). respectively.
Column: A stainless steel column 4.6 mm in inside diame- System repeatability: When the test is repeated 3 times
ter and 6 cm in length, packed with strongly acidic ion-ex- with 10 mL of the standard solution under the above operat-
change resin for liquid chromatography (Na type) composed ing conditions, the relative standard deviations of the peak
with a sulfonated polystyrene copolymer (3 mm in particle di- areas of aspartic acid, proline, valine and arginine are not
ameter). more than 2.0z, respectively.
Peptide content Calculate the peptide content in Calcitonin
579 C.
Salmon by the following equation using amino acid analysis
Chemical reaction bath temperature: A constant tempera-
values (mmol/mL) obtained in the Constituent amino acids:
ture of about 1309C.
it is not less than 80.0z.
Color developing time: About 1 minute.
Mobile phase: Prepare mobile phases A, B, C, D and E Peptide content (z) ＝ 3431.85 × 5/M × A/11 × 100
according to the following table.
A: Total (mmol/mL) of the amino acid analysis values of
valine, leucine, glycine and proline
Mobile Mobile Mobile Mobile Mobile M: Amount (mg) of Calcitonin Salmon taken
phase A phase B phase C phase D phase E
11: Total of the theoretical residue numbers of valine, leu-
Citric acid cine, glycine and proline per one mole of calcitonin
monohydrate 19.80 g 22.00 g 12.80 g 6.10 g —
salmon
Trisodium citrate
dihydrate 6.19 g 7.74 g 13.31 g 26.67 g —
Purity (1) Acetic acid—Weigh accurately about 10 mg of
Sodium hydroxide — — — — 8.00 g Calcitonin Salmon, dissolve in water to make exactly 10 mL,
Sodium chlo- and use this solution as the sample solution. Separately,
ride 5.66 g 7.07 g 3.74 g 54.35 g — weigh accurately about 1 g of acetic acid (100), and dissolve
Ethanol (99.5) 130.0 mL 20.0 mL 4.0 mL — 100.0 mL in water to make exactly 100 mL. Pipet 2 mL of this solu-
Benzyl alcohol — — — 5.0 mL — tion, add water to make exactly 200 mL, and use this solu-
Thiodiglycol 5.0 mL 5.0 mL 5.0 mL — — tion as the standard solution. Perform the test with exactly
Lauromacrogol 100 mL each of the sample solution and standard solution as
solution (1 in 4) 4.0 mL 4.0 mL 4.0 mL 4.0 mL 4.0 mL
Caprylic acid 0.1 mL 0.1 mL 0.1 mL 0.1 mL 0.1 mL the following conditions. Determine the peak areas, AT and
a sufficient a sufficient a sufficient a sufficient a sufficient AS, of acetic acid in each solution, and calculate the amount
Water amount amount amount amount amount of acetic acid by the following equation: the amount of
Total volume 1000 mL 1000 mL 1000 mL 1000 mL 1000 mL acetic acid is not more than 7.0z.
Amount (z) of acetic acid (CH3COOH)
Flowing of mobile phase: Control the gradient by mixing ＝ MS/MT × AT/AS × 1/10
the mobile phases A, B, C, D and E as directed in the follow-
MS: Amount (mg) of acetic acid (100) taken
ing table.
MT: Amount (mg) of Calcitonin Salmon taken
Time after Mobile Mobile Mobile Mobile Mobile Operating conditions—

injection of phase A phase B phase C phase D phase E Detector: An ultraviolet absorption photometer (wave-
sample (min) (volz) (volz) (volz) (volz) (volz) length: 210 nm).
0 – 1.5 100 0 0 0 0 ter and 25 cm in length, packed with octadecylsilanized silica
1.5 – 4 0 100 0 0 0 gel for liquid chromatography (5 mm in particle diameter).
4 – 12 0 0 100 0 0 Column temperature: A constant temperature of about
12 – 26 0 0 0 100 0 409C.
26 – 30 0 0 0 0 100 Mobile phase A: To 0.7 mL of phosphoric acid add 900
mL of water, adjust the pH to 3.0 with 8 mol/L sodium hy-
Reaction reagent: Mix 407 g of lithium acetate dihydrate, droxide TS, and add water to make 1000 mL.
245 mL of acetic acid (100) and 801 mL of 1-methoxy-2- Mobile phase B: Methanol.
propanol, add water to make 2000 mL, stir for 10 minutes Flowing of mobile phase: Control the gradient by mixing
while passing nitrogen, and use this solution as solution A. the mobile phases A and B as directed in the following table.
Separately, to 1957 mL of 1-methoxy-2-propanol add 77 g of
ninhydrin and 0.134 g of sodium borohydride, stir for 30
minutes while passing nitrogen, and use this solution as solu-
tion B. Mix solution A and solution B before use.
Flow rate of mobile phase: About 0.4 mL per minute.
Flow rate of reaction reagent: About 0.35 mL per minute.
JP XVII Official Monographs / Calcitonin Salmon 547
Water <2.48> Not more than 10.0z (5 mg, coulometric

Time after injection Mobile phase A Mobile phase B titration).
of sample (min) (volz) (volz)
Assay (i) Test animals: Select healthy albino rats weigh-
0–5 95 5 ing between 55 and 180 g, fasted for 24 hours before the test
5 – 10 95 → 50 5 → 50 but allowed to drink water ad libitum.
10 – 20 50 50 (ii) Standard solutions: Dissolve a quantity of Calcitonin
20 – 22 50 → 95 50 → 5 Salmon RS in acetic acid buffer solution containing 0.1z
22 – 30 95 5 bovine serum albumin, and prepare a high-dose standard
solution SH and a low-dose standard solution SL containing
Flow rate: Adjust so that the retention time of acetic acid exactly 0.050 and 0.025 Units per mL, respectively.
is about 4 minutes. (iii) Sample solutions: According to the labeled units,
System suitability— weigh accurately a suitable amount of Calcitonin Salmon,
System performance: When the procedure is run with 100 and dissolve in acetic acid buffer solution containing 0.1z
mL of the standard solution under the above operating con- bovine serum albumin, and prepare a high-dose sample solu-
ditions, the number of theoretical plates and the symmetry tion TH and the low-dose sample solution TL having Units
factor of the peak of acetic acid are not less than 3000 and equal to the standard solutions in equal volumes, respec-
not more than 2.0, respectively. tively.
System repeatability: When the test is repeated 6 times (iv) Dose for injection: Inject 0.3 mL per animal.
with 100 mL of the standard solution under the above operat- (v) Procedure: Divide the test animals at random into 4
ing conditions, the relative standard deviation of the peak groups, A, B, C and D, with not less than 8 animals and
areas of acetic acid is not more than 2.0z. equal numbers in each group. Inject SH, SL, TH and TL into
(2) Related substances—Dissolve 2 mg of Calcitonin the tail vein or subcutaneously into the neck of each animal
Salmon in 2 mL of dilute acetic acid, and use this solution as of the respective groups. At 1 hour after the injection, collect
the sample solution. Perform the test with 20 mL of the sam- blood from the abdominal aorta in a way that minimizes the
ple solution as directed under Liquid Chromatography suffering of the animals, allow the blood samples to stand
<2.01> according to the following conditions. Determine each at room temperature for about 30 minutes, and centrifuge
peak area from the sample solution by the automatic integra- at 3000 revolutions per minute for 10 minutes to separate
tion method, and calculate the amount of them by the area serum.
percentage method: the total amount of the peaks other than (vi) Serum calcium determination: Pipet 0.1 mL of the
calcitonin salmon is not more than 3z. serum, add exactly 6.9 mL of strontium TS, mix well, and
Operating conditions— use this solution as the sample solution for calcium determi-
Detector: An ultraviolet absorption photometer (wave- nation. Separately, pipet a suitable volume of Standard Cal-
length: 210 nm). cium Solution for Atomic Absorption Spectrophotometry,
Column: A stainless steel column 3.9 mm in inside diame- dissolve in strontium TS to make a solution so that each mL
ter and 30 cm in length, packed with octadecylsilanized silica contains 0.2 to 3 mg of calcium (Ca: 40.08), and use this
gel for liquid chromatography (10 mm in particle diameter). solution as the standard solution for calcium determination.
Column temperature: A constant temperature of about Perform the test as directed under Atomic Absorption Spec-
259 C. trometry <2.23> according to the following conditions, and
Mobile phase: A mixture of 1z trimethylamine-phosphate calculate the calcium content of the sample solution for
buffer solution (pH 3.0) and acetonitrile (27:13). calcium determination from the calibration curve obtained
Flow rate: Adjust so that the retention time of calcitonin from the absorbance of the standard solution for calcium
salmon is about 9 minutes. determination.
Time span of measurement: About 2 times as long as the Amount (mg) of Calcium (Ca) in 100 mL of the serum
retention time of calcitonin salmon, beginning after the sol- ＝ Calcium content (ppm) in the sample solution for
vent peak. calcium determination × 7
Test for required detectability: To 1 mL of the sample so- Gas: Combustible gas—Acetylene.
lution add the mobile phase to make 100 mL, and use this Supporting gas—Air.
solution as the solution for system suitability test. Pipet 1 Lamp: Calcium hollow-cathode lamp.
mL of the solution for system suitability test, and add the Wavelength: 422.7 nm.
mobile phase to make exactly 10 mL. Confirm that the peak (vii) Calculation: Amounts of calcium in the serum ob-
area of calcitonin salmon obtained from 20 mL of this solu- tained with SH, SL, TH and TL are symbolized as y1, y2, y3
tion is equivalent to 5 to 15z of that obtained from 20 mL of and y4, respectively. Sum up y1, y2, y3 and y4 on each set to
the solution for system suitability test. obtain Y1, Y2, Y3 and Y4, respectively.
System performance: Dissolve 5 mg of methyl parahy-
droxybenzoate and 7 mg of ethyl parahydroxybenzoate in Units per mg of peptide ＝ antilog M × b/a × 1/c × 5
100 mL of acetonitrile. When the procedure is run with 20 M ＝ 0.3010 × (Ya/Yb)
mL of this solution under the above operating conditions Ya ＝－Y1 － Y2 ＋ Y3 ＋ Y4
methyl parahydroxybenzoate and ethyl parahydroxybenzo- Yb ＝ Y1 － Y2 ＋ Y3 － Y4
ate are eluted in this order with the resolution between these
peaks being not less than 5. a: Amount (mg) of Calcitonin Salmon taken
System repeatability: When the test is repeated 6 times b: Total volume (mL) of the high-dose sample solution
with 20 mL of the solution for system suitability test under prepared by dissolving Calcitonin Salmon in acetic acid
the above operating conditions, the relative standard devia- buffer solution containing 0.1z bovine serum albumin.
tion of the peak areas of calcitonin salmon is not more than c: Peptide content (z)
2.0z. F? computed by the following equation should be smaller
548 Precipitated Calcium Carbonate / Official Monographs JP XVII
than F1 shown in the table against n with which s 2 is calcu- nitrate TS, and ignite the residue together with the filter
lated. Calculate L (P ＝ 0.95) by use of the following equa- paper: the mass of the residue is not more than 10.0 mg.
tion: L should be not more than 0.20. If F? exceeds F1, or if (2) Heavy metals <1.07>—Mix 2.0 g of Precipitated Cal-
L exceeds 0.20, repeat the test, increasing the number of cium Carbonate with 5 mL of water, add slowly 6 mL of
animals or arranging the assay conditions so that F? is not dilute hydrochloric acid, and evaporate on a water bath to
more than F1 and L is not more than 0.20. dryness. Dissolve the residue in 50 mL of water, and filter.
To 25 mL of the filtrate add 2 mL of dilute acetic acid, 1
F? ＝ (－Y1 ＋ Y2 ＋ Y3 － Y4)2/4fs 2
drop of ammonia TS and water to make 50 mL, and per-
f: Number of the test animals of each group.
s 2 ＝ {Sy 2 － (Y/f )}/n
the control solution as follows: evaporate 3 mL of hydro-
Sy 2: The sum of squares of y1, y2, y3 and y4 in each group.
chloric acid on a water bath to dryness, and add 2.0 mL of
Y ＝ Y 21 ＋ Y 22 ＋ Y 23 ＋ Y 24
Standard Lead Solution, 2 mL of dilute acetic acid and water
n ＝ 4( f － 1)
to make 50 mL (not more than 20 ppm).
L ＝ 2 (C － 1)(CM 2 ＋ 0.09062)
(3) Barium—Mix 1.0 g of Precipitated Calcium Car-
C ＝ Y 2b/(Y 2b － 4fs 2t 2)
bonate with 10 mL of water, add dropwise 4 mL of hydro-
t 2: Value shown in the following table against n used to
chloric acid with stirring, boil for 5 minutes, cool, add water
calculate s 2.
to make 40 mL, and filter. With the filtrate, perform the test
as directed under Flame Coloration Test <1.04> (1): no green
n t 2 ＝ F1 n t 2 ＝ F1 n t 2 ＝ F1 color appears.
1 161.45 13 4.667 25 4.242 (4) Magnesium and alkali metals—Dissolve 1.0 g of
2 18.51 14 4.600 26 4.225 Precipitated Calcium Carbonate in a mixture of 20 mL of
3 10.129 15 4.543 27 4.210 water and 10 mL of dilute hydrochloric acid, boil, neutralize
4 7.709 16 4.494 28 4.196 with ammonia TS, and add ammonium oxalate TS until
5 6.608 17 4.451 29 4.183 precipitation of calcium oxalate is completed. Heat the mix-
6 5.987 18 4.414 30 4.171 ture on a water bath for 1 hour, cool, dilute with water to
7 5.591 19 4.381 40 4.085 100 mL, shake well, and filter. To 50 mL of the filtrate add
8 5.318 20 4.351 60 4.001 0.5 mL of sulfuric acid, evaporate to dryness, and ignite at
9 5.117 21 4.325 120 3.920 6009C to constant mass: the mass of the residue is not more
10 4.965 22 4.301 ∞ 3.841 than 5.0 mg.
11 4.844 23 4.279 (5) Arsenic <1.11>—Moisten 0.40 g of Precipitated Cal-
12 4.747 24 4.260 cium Carbonate with 1 mL of water, then dissolve in 4 mL
of dilute hydrochloric acid, use this solution as the test solu-
Containers and storage Containers—Hermetic containers. tion, and perform the test (not more than 5 ppm).
Storage—Light-resistant, not exceeding 109C.
4 hours).
Precipitated Calcium Carbonate Assay Weigh accurately about 0.12 g of Precipitated Cal-
cium Carbonate, previously dried, and dissolve in 20 mL of
沈降炭酸カルシウム water and 3 mL of dilute hydrochloric acid. Add 80 mL of
water, 15 mL of a solution of potassium hydroxide (1 in 10)
and 0.05 g of NN indicator, and titrate <2.50> immediately
CaCO3: 100.09
with 0.05 mol/L disodium dihydrogen ethylenediamine
tetraacetate VS until the color of the solution changes from
Precipitated Calcium Carbonate, when dried, con-
red-purple to blue.
tains not less than 98.5z of calcium carbonate
(CaCO3). Each mL of 0.05 mol/L disodium dihydrogen
Description Precipitated Calcium Carbonate occurs as a
＝ 5.005 mg of CaCO3
white, fine crystalline powder. It is odorless and tasteless.
It is practically insoluble in water, but its solubility in Containers and storage Containers—Tight containers.
water is increased in the presence of carbon dioxide.
It is practically insoluble in ethanol (95) and in diethyl
ether. Precipitated Calcium Carbonate
It dissolves with effervescence in dilute acetic acid, in
dilute hydrochloric acid and in dilute nitric acid. Fine Granules
Identification (1) Dissolve 0.5 g of Precipitated Calcium 沈降炭酸カルシウム細粒
Carbonate in 10 mL of dilute hydrochloric acid, boil, then
cool, and neutralize with ammonia TS: the solution responds
to the Qualitative Tests <1.09> for calcium salt. Precipitated Calcium Carbonate Fine Granules con-
(2) Precipitated Calcium Carbonate responds to the tain not less than 95.0z and not more than 105.0z of
Qualitative Tests <1.09> (1) for carbonate. the labeled amount of calcium carbonate (CaCO3:
100.09).
Purity (1) Acid-insoluble substances—To 5.0 g of
Precipitated Calcium Carbonate add 50 mL of water, then
add 20 mL of hydrochloric acid dropwise with stirring, boil ules, with Precipitated Calcium Carbonate.
for 5 minutes, cool, add water to make 200 mL, and filter Identification (1) To a quantity of powdered Precipitated
through filter paper for quantitative analysis. Wash the Calcium Carbonate Fine Granules, equivalent to 0.5 g of
residue until the last washing shows no turbidity with silver Precipitated Calcium Carbonate, add 10 mL of dilute hydro-
JP XVII Official Monographs / Precipitated Calcium Carbonate Tablets 549
chloric acid, shake thoroughly, and filter. Boil the filtrate, 0.12 g of calcium carbonate (CaCO3), add 20 mL of water
then cool, and neutralize with ammonia TS: the solution re- and 3 mL of dilute hydrochloric acid, and agitate for 15
sponds to the Qualitative Tests <1.09> (1), (2) and (3) for cal- minutes with the aid of ultrasonic waves. Then, add 80 mL
cium salt. of water, 15 mL of a solution of potassium hydroxide (1 in
(2) Powdered Precipitated Calcium Carbonate Fine 10) and 50 mg of NN indicator, and titrate <2.50> immedi-
Granules responds to the Qualitative Tests <1.09> (1) for car- ately with 0.05 mol/L disodium dihydrogen ethylenediamine
bonate. tetraacetate VS until the color of the solution changes from
red-purple to blue.
Uniformity of dosage units <6.02> The granules in single-
dose packages meet the requirement of the Mass variation Each mL of 0.05 mol/L disodium dihydrogen
test. ethylenediamine tetraacetate VS
＝ 5.005 mg of CaCO3
tions per minute according to the Paddle method, using 900 Containers and storage Containers—Well-closed contain-
mL of 1st fluid for dissolution test as the dissolution me- ers.
dium, the dissolution rate in 10 minutes of Precipitated Cal-
cium Carbonate Fine Granules is not less than 80z.
Start the test with an accurately weighed amount of Precipitated Calcium Carbonate
Precipitated Calcium Carbonate Fine Granules, equivalent
to about 0.5 g of calcium carbonate (CaCO3), withdraw not Tablets
less than 20 mL of the medium at the specified minute after
沈降炭酸カルシウム錠
starting the test, and filter through a membrane filter with a
pore size not exceeding 0.45 mm. Discard the first 10 mL of
the filtrate, pipet 2 mL of the subsequent filtrate, add the Precipitated Calcium Carbonate Tablets contain not
mobile phase to make exactly 20 mL, and use this solution as less than 95.0z and not more than 105.0z of the
the sample solution. Separately, weigh accurately about 28 labeled amount of calcium carbonate (CaCO3:
mg of calcium carbonate for assay, previously dried at 100.09).
1809C for 4 hours, and dissolve in the dissolution medium to
with Precipitated Calcium Carbonate.
the standard solution. Perform the test with exactly 20 mL Identification (1) To a quantity of powdered Precipitated
each of the sample solution and standard solution as directed Calcium Carbonate Tablets, equivalent to 0.5 g of Precipi-
under Liquid Chromatography <2.01> according to the fol- tated Calcium Carbonate, add 10 mL of dilute hydrochloric
lowing conditions, and determine the peak areas, AT and AS, acid, shake throughly, and filter, if necessary. Boil, then
of calcium in each solution. cool, and neutralize with ammonia TS: the solution responds
to the Qualitative Tests <1.09> (1), (2) and (3) for calcium
salt.
of calcium carbonate (CaCO3)
(2) Powdered Precipitated Calcium Carbonate Tablets
＝ MS/MT × AT/AS × 1/C × 1800
responds to the Qualitative Tests <1.09> (1) for carbonate.
MS: Amount (mg) of calcium carbonate for assay taken
MT: Amount (mg) of Precipitated Calcium Carbonate
Fine Granules taken
C: Labeled amount (mg) of calcium carbonate (CaCO3) in Disintegration <6.09> Apply to the preparation intended to
1g be used as antacid.
Perform the test using the disk: it meets the requirement.
Detector: An electric conductivity detector. Dissolution <6.10> Apply to the preparation intended to be
Column: A polyether ether ketone tube 4.6 mm in inside used as hyperphosphatemia.
diameter and 10 cm in length, packed with slightly acidic When the test is performed at 50 revolutions per minute
ion-exchange silica gel for liquid chromatography (7 mm in according to the Paddle method, using 900 mL of 1st fluid
particle diameter). for dissolution test as the dissolution medium, the dissolu-
Column temperature: A constant temperature of about tion rate in 10 minutes of Precipitated Calcium Carbonate
409 C. Tablets is not less than 80z.
Mobile phase: A mixture of a solution of tartaric acid (3 in Start the test with 1 tablet of Precipitated Calcium Car-
2000) and a solution of dipicolinic acid (1 in 3000) (1:1). bonate Tablets, withdraw not less than 20 mL of the medium
Flow rate: Adjust so that the retention time of calcium is at the specified minute after starting the test, and filter
about 8 minutes. through a membrane filter with a pore size not exceeding
System suitability— 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL
System performance: When the procedure is run with 20 of the subsequent filtrate, add the mobile phase to make ex-
mL of the standard solution under the above operating con- actly V? mL so that each mL contains about 56 mg of calcium
ditions, sodium and calcium are eluted in this order with the carbonate (CaCo3), and use this solution as the sample solu-
resolution between these peaks being not less than 4.5. tion. Separately, weigh accurately about 28 mg of calcium
System repeatability: When the test is repeated 5 times carbonate for assay, previously dried at 1809C for 4 hours,
with 20 mL of the standard solution under the above operat- and dissolve in the dissolution medium to make exactly 100
ing conditions, the relative standard deviation of the peak mL. Pipet 10 mL of this solution, add the mobile phase to
area of calcium is not more than 2.0z. make exactly 50 mL, and use this solution as the standard
Assay Weigh accurately a quantity of powdered Precipitat-
ed Calcium Carbonate Fine Granules, equivalent to about
550 Calcium Chloride Hydrate / Official Monographs JP XVII
ditions, and determine the peak areas, AT and AS, of calcium Calcium Chloride Hydrate
in each solution.
塩化カルシウム水和物
of calcium carbonate (CaCO3)
＝ MS × AT/AS × V?/V × 1/C × 180 CaCl2.2H2O: 147.01
MS: Amount (mg) of calcium carbonate for assay taken
Calcium Chloride Hydrate contains not less than
C: Labeled amount (mg) of calcium carbonate (CaCO3) in
96.7z and not more than 103.3z of calcium chloride
1 tablet
hydrate (CaCl2.2H2O).
Description Calcium Chloride Hydrate occurs as white,
Detector: An electric conductivity detector.
granules or masses. It is odorless.
Column: A polyether ether ketone tube 4.6 mm in inside
It is very soluble in water, and soluble in ethanol (95), and
diameter and 10 cm in length, packed with slightly acidic
practically insoluble in diethyl ether.
ion-exchange silica gel for liquid chromatography (7 mm in
It is deliquescent.
particle diameter).
Column temperature: A constant temperature of about Identification A solution of Calcium Chloride Hydrate
409 C. (1 in 10) responds to the Qualitative Tests <1.09> for calcium
Mobile phase: A mixture of a solution of tartaric acid (3 in salt and for chloride.
2000) and a solution of dipicolinic acid (1 in 3000) (1:1).
pH <2.54> The pH of a solution of 1.0 g of Calcium Chlo-
Flow rate: Adjust so that the retention time of calcium is
ride Hydrate in 20 mL of freshly boiled and cooled water is
about 8 minutes.
System performance: When the procedure is run with 20 Purity (1) Clarity and color of solution—A solution of
mL of the standard solution under the above operating con- 1.0 g of Calcium Chloride Hydrate in 20 mL of water is clear
ditions, sodium and calcium are eluted in this order with the and colorless.
resolution between these peaks being not less than 4.5. (2) Sulfate <1.14>—Take 1.0 g of Calcium Chloride Hy-
System repeatability: When the test is repeated 5 times drate, and perform the test. Prepare the control solution
with 20 mL of the standard solution under the above operat- with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more
ing conditions, the relative standard deviation of the peak than 0.024z).
area of calcium is not more than 2.0z. (3) Hypochlorite—Dissolve 0.5 g of Calcium Chloride
Hydrate in 5 mL of water, add 2 to 3 drops of dilute hydro-
Acid-neutralizing capacity <6.04> Apply to the preparation
chloric acid and 2 to 3 drops of zinc iodide-starch TS: no
intended to be used as antacid.
blue color develops immediately.
Weigh accurately and powder not less than 40 Precipitated
(4) Heavy metals <1.07>—Proceed with 2.0 g of Calcium
Calcium Carbonate Tablets. Perform the test with an accu-
Chloride Hydrate according to Method 1, and perform the
rately weighed amount of the powder, equivalent to about
0.25 g of Calcium Carbonate: the amount of 0.1 mol/L hy-
drochloric acid VS consumed per 1 g of Precipitated Cal-
(5) Iron, aluminum or phosphate—Dissolve, in a Nessler
cium Carbonate is not less than 190 mL.
tube, 1.0 g of Calcium Chloride Hydrate in 20 mL of water
Assay Weigh accurately and powder not less than 20 and 1 drop of dilute hydrochloric acid, boil, then cool, add
Precipitated Calcium Carbonate Tablets. To an accurately 3 drops of ammonia TS, and heat the solution to boil: no
weighed portion of the powder, equivalent to about 0.12 g of turbidity or precipitate is produced.
calcium carbonate (CaCO3), add 20 mL of water, 3 mL of (6) Barium—Dissolve 0.5 g of Calcium Chloride Hydrate
dilute hydrochloric acid, and agitate, if necessary, for 15 in 5 mL of water, add 2 drops of dilute hydrochloric acid
minutes with the aid of ultrasonic waves. Then, add 80 mL and 2 mL of potassium sulfate TS, and allow to stand for 10
of water, 15 mL of a solution of potassium hydroxide (1 in minutes: no turbidity is produced.
10) and 50 mg of NN indicator, and titrate <2.50> immedi- (7) Arsenic <1.11>—Prepare the test solution with 1.0 g
ately with 0.05 mol/L disodium dihydrogen ethylenediamine of Calcium Chloride Hydrate according to Method 1, and
tetraacetate VS until the color of the solution changes from perform the test (not more than 2 ppm).
red-purple to blue.
Assay Weigh accurately about 0.4 g of Calcium Chloride
Each mL of 0.05 mol/L disodium dihydrogen Hydrate, and dissolve in water to make exactly 200 mL.
ethylenediamine tetraacetate VS Measure exactly 20 mL of this solution, add 40 mL of water,
＝ 5.005 mg of CaCO3 2 mL of 8 mol/L potassium hydroxide TS and 0.1 g of NN
indicator, and titrate <2.50> immediately with 0.02 mol/L
disodium dihydrogen ethylenediamine tetraacetate VS until
the color of the solution changes from red-purple to blue.
＝ 2.940 mg of CaCl2.2H2O
JP XVII Official Monographs / Calcium Folinate 551
It is sparingly soluble in water, and practically insoluble in

Calcium Chloride Injection methanol and in ethanol (99.5).
塩化カルシウム注射液
solution of Calcium Folinate (1 in 100,000) as directed under
Calcium Chloride Injection is an aqueous injection. the spectrum with the Reference Spectrum or the spectrum
It contains not less than 95.0z and not more than of a solution of Calcium Folinate RS prepared in the same
105.0z of the labeled amount of calcium chloride manner as the sample solution: both spectra exhibit similar
(CaCl2: 110.98). intensities of absorption at the same wavelengths.
The concentration of Calcium Chloride Injection is (2) Determine the infrared absorption spectrum of Cal-
expressed as the quantity of calcium chloride (CaCl2). cium Folinate as directed in the potassium bromide disk
tion, with Calcium Chloride Hydrate.
Description Calcium Chloride Injection is a clear, colorless numbers.
liquid. (3) A solution of Calcium Folinate (1 in 100) responds to
the Qualitative Tests <1.09> (2) and (3) for calcium salt.
Identification Calcium Chloride Injection responds to the
Qualitative Tests <1.09> for calcium salt and for chloride. Optical rotation <2.49> [a]20
D : ＋14 – ＋199(0.1 g calculated
pH <2.54> 4.5 – 7.5
pH <2.54> To 1.25 g of Calcium Folinate add 50 mL of
freshly boiled and cooled water, and warm to 409C, if neces-
Extractable volume <6.05> It meets the requirement. sary, to dissolve: the pH of this solution is between 6.8 and
8.0.
to Method 1: it meets the requirement. Purity (1) Clarity and color of solution—To 1.25 g of
Calcium Folinate add 50 mL of freshly boiled and cooled
water, and warm to 409 C, if necessary, to dissolve: the solu-
ment.
tion is clear, and the absorbance at 420 nm of it, determined
Sterility <4.06> Perform the test according to the Mem- as directed under Ultraviolet-visible Spectrophotometry
brane filtration method: it meets the requirement. <2.24>, is not more than 0.25.
(2) Heavy metals <1.07>—Proceed with 0.40 g of Cal-
Assay Measure exactly a volume of Calcium Chloride
cium Folinate according to Method 2, and perform the test.
Injection, equivalent to about 0.4 g of calcium chloride
(CaCl2), and proceed as directed in the Assay under Calcium
Chloride Hydrate.
(3) Related substances—Dissolve 10 mg of Calcium
Each mL of 0.02 mol/L disodium dihydrogen Folinate in 25 mL of water, and use this solution as the sam-
ethylenediamine tetraacetate VS ple solution. Pipet 2 mL of the sample solution, add water to
＝ 2.220 mg of CaCl2 make exactly 200 mL, and use this solution as the standard
tegration method: the area of the peak other than folinate
Calcium Folinate obtained from the sample solution is not larger than the peak
area of folinate obtained from the standard solution, and the
Calcium Leucovorin total area of the peaks other than folinate from the sample
solution is not larger than 5 times the peak area of folinate
ホリナートカルシウム
the Assay.
retention time of folinate, beginning after the solvent peak.
C20H21CaN7O7: 511.50 System performance: Proceed as directed in the system
Monocalcium N-(4-{[(2-amino-5-formyl-4-oxo-1,4,5,6,7,8- suitability in the Assay.
hexahydropteridin-6-yl)methyl]amino}benzoyl)-L-glutamate Test for required detectability: Pipet 5 mL of the standard
[1492-18-8] solution, and add water to make exactly 50 mL. Confirm
that the peak area of folinate obtained from 20 mL of this so-
Calcium Folinate contains not less than 95.0z lution is equivalent to 7 to 13z of that obtained from 20 mL
and not more than 102.0z of calcium folinate of the standard solution.
(C20H21CaN7O7), calculated on the anhydrous basis. System repeatability: When the test is repeated 6 times
Description Calcium Folinate occurs as a white to light yel-
low crystalline powder.
area of folinate is not more than 2.0z.
552 Calcium Gluconate Hydrate / Official Monographs JP XVII
Water <2.48> Not less than 7.0z and not more than 17.0z (99.5).
Identification (1) To separately 10 mg each of Calcium
Assay Weigh accurately about 10 mg each of Calcium Gluconate Hydrate and calcium gluconate for thin-layer
Folinate and Calcium Folinate RS (separately determine the chromatography add 1 mL of water, dissolve by warming,
water <2.48> in the same manner as Calcium Folinate), dis- and use these solutions as the sample solution and the stand-
solve in water to make them exactly 25 mL. Pipet 5 mL each ard solution, respectively. Perform the test with these solu-
of these solutions, add the mobile phase to make exactly 25 tions as directed under Thin-layer Chromatography <2.03>.
mL, and use these solutions as the sample solution and the Spot 5 mL each of the sample solution and standard solution
standard solution, respectively. Perform the test with exactly on a plate of silica gel for thin-layer chromatography. De-
20 mL each of the sample solution and standard solution as velop the plate with a mixture of ethanol (95), water, ammo-
directed under Liquid Chromatography <2.01> according to nia solution (28) and ethyl acetate (5:3:1:1) to a distance of
the following conditions, and determine the peak areas, AT about 10 cm, air-dry the plate, and heat the plate at 1109C
and AS, of folinate in each solution. for 20 minutes. After cooling, spray evenly hexaammonium
heptamolybdate-cerium (IV) sulfate TS on the plate, air-dry,
Amount (mg) of calcium folinate (C20H21CaN7O7)
and heat at 1109 C for 10 minutes: the spots with the sample
＝ M S × AT / AS
solution and the standard solution are the same in the R f
MS: Amount (mg) of Calcium Folinate RS taken, calcu- value and color tone.
lated on the anhydrous basis (2) A solution of Calcium Gluconate Hydrate (1 in 40)
responds to the Qualitative Tests <1.09> (1), (2) and (3) for
calcium salt.
length: 254 nm). Optical rotation <2.49> [a]20D : ＋6 – ＋119 (after drying,
Column: A stainless steel column 4.6 mm in inside diame- 0.5 g, water, warming, after cooling, 25 mL, 100 mm).
pH <2.54> Dissolve 1.0 g of Calcium Gluconate Hydrate in
20 mL of water by warming: the pH of the solution is be-
tween 6.0 and 8.0.
459 C.
Mobile phase: A mixture of disodium hydrogen phosphate Purity (1) Clarity and color of solution—Dissolve 1.0 g
dodecahydrate solution (287 in 100,000), methanol and of Calcium Gluconate Hydrate in 50 mL of water by warm-
tetrabutylammonium hydroxide TS (385:110:4), adjusted to ing: the solution is clear and colorless.
pH7.5 with phosphoric acid. (2) Chloride <1.03>—Take 0.40 g of Calcium Gluconate
Flow rate: Adjust so that the retention time of folinate is Hydrate, and perform the test. Prepare the control solution
about 10 minutes. with 0.80 mL of 0.01 mol/L hydrochloric acid VS (not more
System suitability— than 0.071z).
System performance: Dissolve 10 mg each of Calcium (3) Sulfate <1.14>—Take 1.0 g of Calcium Gluconate
Folinate and folic acid in 100 mL of the mobile phase. When Hydrate, and perform the test. Prepare the control solution
the procedure is run with 20 mL of this solution under the with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than
above operating conditions, folinate and folic acid are eluted 0.048z).
in this order with the resolution between these peaks being (4) Heavy metals <1.07>—Dissolve 1.0 g of Calcium
not less than 10. Gluconate Hydrate in 30 mL of water and 2 mL of dilute
System repeatability: When the test is repeated 6 times acetic acid by warming, cool, and add water to make 50 mL.
with 20 mL of the standard solution under the above operat- Perform the test using this solution as the test solution. Pre-
ing conditions, the relative standard deviation of the peak pare the control solution with 2.0 mL of Standard Lead So-
area of folinate is not more than 1.0z. lution, 2 mL of dilute acetic acid, and water to make 50 mL
(5) Arsenic <1.11>—Dissolve 0.6 g of Calcium Gluconate
Hydrate in 5 mL of water by warming, add 5 mL of dilute
sulfuric acid and 1 mL of bromine TS, and concentrate on a
water bath to 5 mL. Perform the test with this solution as the
Calcium Gluconate Hydrate test solution (not more than 3.3 ppm).
(6) Sucrose and reducing sugars—To 0.5 g of Calcium
グルコン酸カルシウム水和物
Gluconate Hydrate add 10 mL of water and 2 mL of dilute
hydrochloric acid, and boil the solution for 2 minutes. After
cooling, add 5 mL of sodium carbonate TS, allow to stand
for 5 minutes, add water to make 20 mL, and filter. To 5 mL
of the filtrate add 2 mL of Fehling's TS, and boil for 1
minute: no orange-yellow to red precipitate is formed imme-
C12H22CaO14.H2O: 448.39
diately.
Monocalcium di-D-gluconate monohydrate
[299-28-5] Loss on drying <2.41> Not more than 1.0z (1 g, 809C,
2 hours).
Calcium Gluconate Hydrate, when dried, contains
Assay Weigh accurately about 0.4 g of Calcium Gluconate
not less than 99.0z and not more than 104.0z of cal-
Hydrate, previously dried, dissolve in 100 mL of water, add
cium gluconate hydrate (C12H22CaO14.H2O).
2 mL of 8 mol/L potassium hydroxide TS and 0.1 g of NN
Description Calcium Gluconate Hydrate occurs as a white, indicator, and titrate <2.50> immediately with 0.05 mol/L
crystalline powder or granules. disodium dihydrogen ethylenediamine tetraacetate VS until
It is soluble in water, and practically insoluble in ethanol the color of the solution changes from red-purple to blue.
JP XVII Official Monographs / Calcium Lactate Hydrate 553
Each mL of 0.05 mol/L disodium dihydrogen add water to make exactly 100 mL. Measure exactly 10 mL
ethylenediamine tetraacetate VS of this solution, add 90 mL of water and 1.5 mL of 8 mol/L
＝ 22.42 mg of C12H22CaO14.H2O potassium hydroxide TS, shake, allow to stand for 3 to 5
minutes, and then add 0.1 g of NN indicator. Titrate <2.50>
immediately with 0.05 mol/L disodium dihydrogen ethylene-
ers.
diamine tetraacetate VS, until the red-purple color of the so-
lution changes to blue.
Calcium Hydroxide Each mL of 0.05 mol/L disodium dihydrogen

Slaked Lime ＝ 3.705 mg of Ca(OH)2
水酸化カルシウム
Ca(OH)2: 74.09 Calcium Lactate Hydrate

Calcium Hydroxide contains not less than 90.0z of 乳酸カルシウム水和物
calcium hydrate [Ca(OH)2].
Description Calcium Hydroxide occurs as a white powder.
It has a slightly bitter taste.
It is slightly soluble in water, very slightly soluble in boil-
C6H10CaO6.5H2O: 308.29
ing water, and practically insoluble in ethanol (95) and in
Monocalcium bis[(2RS )-2-hydroxypropanonate]
diethyl ether.
pentahydrate
It dissolves in dilute acetic acid, in dilute hydrochloric acid
[63690-56-2]
and in dilute nitric acid.
It absorbs carbon dioxide from air.
Calcium Lactate Hydrate, when dried, contains not
Identification (1) Mix Calcium Hydroxide with 3 to 4 less than 97.0z of calcium lactate (C6H10CaO6:
times its mass of water: the mixture is slushy and is alkaline. 218.22).
(2) Dissolve 1 g of Calcium Hydroxide in 30 mL of dilute
Description Calcium Lactate Hydrate occurs as white,
acetic acid, and boil. After cooling, neutralize with ammonia
powder or granules. It is odorless, and has a slightly acid
TS: the solution responds to the Qualitative tests <1.09> (2)
taste.
and (3) for calcium salt.
A 1 g portion of it dissolves gradually in 20 mL of water,
Purity (1) Acid-insoluble substances—To 5 g of Calcium and it is slightly soluble in ethanol (95), and practically in-
Hydroxide add 100 mL of water, add hydrochloric acid soluble in diethyl ether.
dropwise with strring until the solution becomes acidic, and It is partly efflorescent at ordinary temperature, and yields
further add 1 mL of hydrochloric acid. Boil this solution for the anhydride at 1209C.
5 minutes, cool, and filter through a tared glass filter (G4).
Identification A solution of Calcium Lactate Hydrate (1 in
Wash the residue with boiling water until the last washing
20) responds to the Qualitative Tests <1.09> for calcium salt
exhibits no turbidity upon addition of silver nitrate TS, and
and for lactate.
dry at 1059C to constant mass: the mass is not more than 25
mg. Purity (1) Clarity of solution—Dissolve 1.0 g of Calcium
(2) Heavy metals <1.07>—Dissolve 1.0 g of Calcium Hy- Lactate Hydrate in 20 mL of water by warming: the solution
droxide in 10 mL of dilute hydrochloric acid, evaporate on a is clear.
water bath to dryness, dissolve the residue in 40 mL of (2) Acidity or alkalinity—To the solution obtained in (1)
water, and filter. To 20 mL of the filtrate add 2 mL of dilute add 2 drops of phenolphthalein TS: no red color is pro-
acetic acid and water to make 50 mL, and perform the test duced. Then add 0.50 mL of 0.1 mol/L sodium hydroxide
using this solution as the test solution. Prepare the control VS: a red color develops.
solution as follows: evaporate 5 mL of dilute hydrochloric (3) Heavy metals <1.07>—Dissolve 1.0 g of Calcium Lac-
acid on a water bath to dryness, and add 2 mL of dilute tate Hydrate in 30 mL of water and 5 mL of dilute acetic
acetic acid, 2.0 mL of Standard Lead Solution and water to acid by warming, cool, add water to make 50 mL, and per-
make 50 mL (not more than 40 ppm). form the test using this solution as the test solution. Prepare
(3) Magnesium and alkali metals—Dissolve 1.0 g of Cal- the control solution from 2.0 mL of Standard Lead Solution
cium Hydroxide in a mixture of 20 mL of water and 10 mL and 2 mL of dilute acetic acid, and dilute with water to 50
of dilute hydrochloric acid, boil, neutralize with ammonia mL (not more than 20 ppm).
TS, and precipitate calcium oxalate completely by adding (4) Magnesium or alkali metals—Dissolve 1.0 g of Cal-
dropwise ammonium oxalate TS. Heat the mixture on a cium Lactate Hydrate in 40 mL of water, add 0.5 g of am-
water bath for 1 hour, cool, dilute with water to 100 mL, monium chloride, boil, then add 20 mL of ammonium oxa-
shake, and filter. To 50 mL of the filtrate add 0.5 mL of sul- late TS. Heat the mixture on a water bath for 1 hour, cool,
furic acid, evaporate to dryness, and ignite at 6009C to con- dilute with water to 100 mL, and filter. To 50 mL of the fil-
stant mass: the mass of the residue does not exceed 24 mg. trate add 0.5 mL of sulfuric acid, evaporate to dryness, and
(4) Arsenic <1.11>—Dissolve 0.5 g of Calcium Hydroxide ignite between 4509C and 5509C to constant mass: the mass
in 5 mL of dilute hydrochloric acid, and perform the test of the residue is not more than 5 mg.
with this solution as the test solution (not more than 4 ppm). (5) Arsenic <1.11>—Dissolve 0.5 g of Calcium Lactate
Hydrate in 2 mL of water and 3 mL of hydrochloric acid,
Assay Weigh accurately about 1 g of Calcium Hydroxide,
and perform the test with this solution as the test solution
dissolve by adding 10 mL of dilute hydrochloric acid, and
554 Calcium Oxide / Official Monographs JP XVII
(not more than 4 ppm). for 1 to 2 minutes, neutralize with ammonia TS, add drop-
(6) Volatile fatty acid—Warm 1.0 g of Calcium Lactate wise an excess of hot ammonium oxalate TS, heat the mix-
Hydrate with 2 mL of sulfuric acid: an odor of acetic acid or ture on a water bath for 2 hours, cool, add water to make
butyric acid is not perceptible. 200 mL, mix thoroughly, and filter. Evaporate 50 mL of the
filtrate with 0.5 mL of sulfuric acid to dryness, and heat the
Loss on drying <2.41> 25.0 – 30.0z (1 g, 809C, 1 hour at
residue strongly at 6009C to constant mass: the mass of the
first, then 1209C, 4 hours).
residue is not more than 15 mg.
Assay Weigh accurately about 0.5 g of Calcium Lactate
Loss on ignition <2.43> Not more than 10.0z (1 g, 9009C,
Hydrate, previously dried, add water, dissolve by heating on
constant mass).
a water bath, cool, and add water to make exactly 100 mL.
Pipet 20 mL of this solution, then 80 mL of water and 1.5 Assay Weigh accurately about 0.7 g of Calcium Oxide, pre-
mL of 8 mol/L potassium hydroxide TS, and allow to stand viously incinerated at 9009C to constant mass and cooled in
for 3 to 5 minutes. Add 0.1 g of NN indicator, and titrate a desiccator (silica gel), and dissolve in 50 mL of water and 8
<2.50> immediately with 0.02 mol/L disodium dihydrogen mL of diluted hydrochloric acid (1 in 3) by heating. Cool,
ethylenediamine tetraacetate VS until the color of the solu- and add water to make exactly 250 mL. Pipet 10 mL of the
tion changes from red to blue. solution, add 50 mL of water, 2 mL of 8 mol/L potassium
hydroxide TS and 0.1 g of NN indicator, and titrate <2.50>
with 0.02 mol/L disodium dihydrogen ethylenediamine
tetraacetate VS, until the red-purple color of the solution
＝ 4.364 mg of C6H10CaO6
changes to blue.
＝ 1.122 mg of CaO
Calcium Oxide
Quick Lime
酸化カルシウム Calcium Pantothenate
パントテン酸カルシウム
CaO: 56.08
Calcium Oxide, when incinerated, contains not less

than 98.0z of calcium oxide (CaO).
Description Calcium Oxide occurs as hard, white masses,
containing a powder. It is odorless. C18H32CaN2O10: 476.53
It is very slightly soluble in boiling water, and practically Monocalcium bis{3-[(2R)-2,4-dihydroxy-3,3-
insoluble in ethanol (95). dimethylbutanoylamino]propanoate}
One gram of Calcium Oxide dissolves almost completely [137-08-6]
in 2500 mL of water.
It slowly absorbs moisture and carbon dioxide from air. Calcium Pantothenate contains not less than 98.0z
and not more than 102.0z of calcium pantothenate
Identification (1) Moisten Calcium Oxide with water:
(C18H32CaN2O10), calculated on the dried basis.
heat is generated and a white powder is obtained. Mix the
powder with about 5 times its mass of water: the mixture is Description Calcium Pantothenate occurs as a white pow-
alkaline. der.
(2) Dissolve 1 g of Calcium Oxide in 20 mL of water by It is freely soluble in water, and practically insoluble in
adding a few drops of acetic acid (31): the solution responds ethanol (99.5).
to the Qualitative Tests <1.09> for calcium salt. The pH of a solution prepared by dissolving 1.0 g of Cal-
cium Pantothenate in 20 mL of water is between 7.0 and 9.0.
Purity (1) Acid-insoluble substances—Disintegrate 5.0 g
It is hygroscopic.
of Calcium Oxide with a small amount of water, add 100 mL
of water, add dropwise hydrochloric acid with stirring until
the solution becomes acidic, and further add 1 mL of hydro- Identification (1) Determine the infrared absorption spec-
chloric acid. Boil the solution for 5 minutes, cool, filter trum of previously dried Calcium Pantothenate as directed
through a glass filter (G4), wash the residue with boiling in the potassium bromide disk method under Infrared Spec-
water until no turbidity is produced when silver nitrate TS is trophotometry <2.25>, and compare the spectrum with the
added to the last washing, and dry at 1059 C to constant Reference Spectrum or the spectrum of dried Calcium
mass: the mass of the residue is not more than 10.0 mg. Pantothenate RS: both spectra exhibit similar intensities of
(2) Carbonate—Disintegrate 1.0 g of Calcium Oxide absorption at the same wave numbers. If any difference
with a small amount of water, mix thoroughly with 50 mL of appears between the spectra, dissolve the sample and the
water, allow to stand for a while, remove most of the super- Reference Standard separately in water, evaporate water,
natant milky liquid by decantation, and add an excess of dry the residues in vacuum for 24 hours using silica gel as a
dilute hydrochloric acid to the residue: no vigorous efferves- desiccant, and perform the test using these residues.
cence is produced. (2) A solution of Calcium Pantothenate (1 in 10) re-
(3) Magnesium and alkali metals—Dissolve 1.0 g of Cal- sponds to the Qualitative Tests <1.09> (1), (2) and (3) for cal-
cium Oxide in 75 mL of water by adding dropwise hydro- cium salt.
chloric acid, and further add 1 mL of hydrochloric acid. Boil
JP XVII Official Monographs / Calcium Paraaminosalicylate Hydrate 555
Optical rotation <2.49> [a]20D : ＋25.0 – ＋28.59(1 g calcu- 100 mL, and use these solutions as the sample solution and
lated on the dried basis, water, 20 mL, 100 mm). the standard solution, respectively. Perform the test with ex-
Calcium Pantothenate according to Method 1, and perform
cording to the following conditions, and determine the peak
areas, AT and AS, of pantothenic acid in each solution.
(2) Related substances—Dissolve 0.30 g of Calcium Pan- Amount (mg) of calcium pantothenate (C18H32CaN2O10)
tothenate in 20 mL of water, and use this solution as the ＝ M S × AT / AS
MS: Amount (mg) of Calcium Pantothenate RS taken,
calculated on the dried basis
of the sample solution and standard solution as directed Operating conditions—
under Liquid Chromatography <2.01> according to the fol- Detector: An ultraviolet absorption photometer (wave-
lowing conditions, and determine each peak area by the au- length: 210 nm).
tomatic integration method: the area of the peak, having the Column: A stainless steel column 4.6 mm in inside diame-
relative retention time of about 0.6 to pantothenic acid ob- ter and 25 cm in length, packed with octadecylsilanized silica
tained from the sample solution is not larger than 1.2 times gel for liquid chromatography (5 mm in particle diameter).
the peak area of pantothenic acid obtained from the stand- Column temperature: A constant temperature of about
ard solution, the area of the peak, having the relative reten- 409C.
tion time of about 0.8 is not larger than the peak area of Mobile phase: Dissolve 0.81 g of sodium 1-heptanesul-
pantothenic acid from the standard solution, the area of the fonate and 1.36 g of potassium dihydrogen phosphate in
peak, having the relative retention time of about 1.5 is not water to make 1000 mL, and adjust to pH 2.1 with phos-
larger than 3/5 times the peak area of pantothenic acid from phoric acid. To 980 mL of this solution add 10 mL of aceto-
the standard solution, and the area of the peak other than nitrile and 10 mL of methanol.
pantothenic acid and the peaks mentioned above is not Flow rate: Adjust so that the retention time of pantothenic
larger than 3/10 times the peak area of pantothenic acid acid is about 17 minutes.
from the standard solution. Additionally, the total area of System suitability—
the peaks other than pantothenic acid from the sample solu- System performance: When the procedure is run with 10
tion is not larger than 2.4 times the peak area of pantothenic mL of the standard solution under the above operating con-
acid from the standard solution. For the areas of the peaks, ditions, the number of theoretical plates and the symmetry
having the relative retention time of about 0.6 and about 0.8 factor of the peak of pantothenic acid are not less than
to pantothenic acid, multiply their relative response factors, 10,000 and not more than 1.5, respectively.
19 and 13, respectively. System repeatability: When the test is repeated 6 times
Operating conditions— with 10 mL of the standard solution under the above operat-
Detector, column, column temperature, mobile phase, and ing conditions, the relative standard deviation of the peak
flow rate: Proceed as directed in the operating conditions in area of pantothenic acid is not more than 1.0z.
the Assay.
retention time of pantothenic acid, beginning after the sol-
vent peak.
System suitability— Calcium Paraaminosalicylate
Test for required detectability: To exactly 2 mL of the Hydrate
standard solution add water to make exactly 10 mL. Con-
firm that the peak area of pantothenic acid obtained with 10 Pas-calcium Hydrate
mL of this solution is equivalent to 14 to 26z of that with 10
mL of the standard solution. パラアミノサリチル酸カルシウム水和物
factor of the peak of pantothenic acid are not less than
10,000 and not more than 1.5, respectively.
C7H5CaNO3･3 1/2 H2O: 254.25
Monocalcium 4-amino-2-oxidobenzoate hemiheptahydrate
area of pantothenic acid is not more than 2.0z.
(3) Alkaloids—Dissolve 50 mg of Calcium Pantothenate
in 5 mL of water, add 0.5 mL of hexaammonium heptamo-
Calcium Paraaminosalicylate Hydrate contains not
lybdate TS and 0.5 mL of a solution of phosphoric acid (1 in
less than 97.0z and not more than 103.0z of calcium
10): no white turbidity is produced.
paraaminosalicylic acid (C7H5CaNO3: 191.20), calcu-
Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, lated on the anhydrous basis.
4 hours).
Description Calcium Paraaminosalicylate Hydrate occurs
Assay Weigh accurately about 20 mg each of Calcium Pan- as a white to slightly colored powder. It has a slightly bitter
tothenate and Calcium Pantothenate RS (separately deter- taste.
mine the loss on drying <2.41> in the same conditions as Cal- It is very slightly soluble in water, and practically insoluble
cium Pantothenate), dissolve each in water to make exactly in methanol and in ethanol (99.5).
556 Calcium Paraaminosalicylate Granules / Official Monographs JP XVII
It is gradually colored to brown by light. mL of potassium iodide TS, and shake gently. After 5
minutes, titrate <2.50> the produced iodine with 0.1 mol/L
Identification (1) To 50 mg of Calcium Paraaminosalicy-
sodium thiosulfate VS (indicator: 1 mL of starch TS). Per-
late Hydrate add 100 mL of water, shake well, and filter. To
form a blank determination.
10 mL of the filtrate add 1 mL of 1 mol/L hydrochloric acid
TS, shake, and add 1 drop of iron (III) chloride TS: a red- Each mL of 0.05 mol/L bromine VS
purple color develops. ＝ 3.187 mg of C7H5CaNO3
(2) Determine the infrared absorption spectrum of Cal-
cium Paraaminosalicylate Hydrate as directed in the potas-
tion at the same wave numbers. Calcium Paraaminosalicylate
(3) To 3 g of Calcium Paraaminosalicylate Hydrate add Granules
15 mL of ammonium chloride TS and 15 mL of water, heat
on a water bath until almost dissolved, and filter after cool- Pas-calcium Granules
ing: the filtrate responds to the Qualitative Tests <1.09> (1),
(2) and (3) for calcium salt. パラアミノサリチル酸カルシウム顆粒
Purity (1) Chloride <1.03>—Dissolve 1.0 g of Calcium
Paraaminosalicylate Hydrate in 15 mL of dilute nitric acid, Calcium Paraaminosalicylate Granules contain not
and add water to make 50 mL. Perform the test using this less than 95.0z and not more than 105.0z of the
solution as the test solution. Prepare the control solution labeled amount of calcium paraaminosalicylate hy-
with 0.70 mL of 0.01 mol/L hydrochloric acid VS (not more drate (C7H5CaNO3.31/2H2O: 254.25).
than 0.025z).
ules, with Calcium Paraaminosalicylate Hydrate.
Paraaminosalicylate Hydrate according to method 3, and
perform the test. Prepare the control solution with 2.0 mL of Identification Powder Calcium Paraaminosalicylate Gran-
Standard Lead Solution (not more than 20 ppm). ules, weigh a portion of the powder, equivalent to 50 mg of
(3) Arsenic <1.11>—Dissolve 0.40 g of Calcium Para- Calcium Paraaminosalicylate Hydrate, add 100 mL of
aminosalicylate Hydrate in 20 mL of 0.1 mol/L hydrochloric water, shake, and filter. To 10 mL of the filtrate add 1 mL
acid TS by warming on a water bath, use this solution as the of 1 mol/L hydrochloric acid TS, shake, and add 1 drop of
test solution, and perform the test (not more than 5 ppm). iron (III) chloride TS: a red-purple color develops.
(4) 3-Aminophenol—To 0.10 g of Calcium Para-
aminosalicylate Hydrate add 5 mL of 0.1 mol/L disodium
dihydrogen ethylenediamine tetraacetate TS, previously
cooled in ice-water, and dissolve by shaking vigorously. Add
in 60 minutes of Calcium Paraaminosalicylate Granules is
immediately 3 mL of ammonia-ammonium chloride buffer
not less than 75z.
solution (pH 11.0) previously cooled in ice water, and shake.
Start the test with an accurately weighed amount of Cal-
Add 2 mL of 4-amino-N, N-diethylaniline sulfate TS, shake,
cium Paraaminosalicylate Granules, equivalent to about
add 10.0 mL of cyclohexane and 4 mL of diluted potassium
0.25 g of calcium paraaminosalicylate hydrate (C7H5CaNO3.
hexacyanoferrate (III) TS (1 in 10), and shake immediately
3 1/2 H2O), withdraw not less than 20 mL of the medium at
for 20 seconds. Centrifuge this solution, wash the separated
cyclohexane layer with two 5-mL portions of diluted ammo-
nia TS (1 in 14), add 1 g of anhydrous sodium sulfate, shake,
Discard the first 10 mL of the filtrate, pipet 5 mL of the sub-
and allow to stand for 5 minutes: the clear cyclohexane layer
sequent filtrate, add water to make exactly 100 mL, and use
is not more colored than the following control solution.
Control solution: Dissolve 50 mg of 3-aminophenol in
rately about 28 mg of calcium paraaminosalicylate hydrate
water, and dilute with water to exactly 500 mL. Measure ex-
for assay (separately determine the water <2.48> in the same
actly 20 mL of this solution, and add water to make exactly
manner as Calcium Paraaminosalicylate Hydrate), and dis-
100 mL. Take 5.0 mL of this solution, add 3 mL of ammo-
solve in water to make exactly 100 mL. Pipet 5 mL of this
nia-ammonium chloride buffer solution (pH 11.0) previously
solution, add water to make exactly 100 mL, and use this so-
cooled in ice-water, and treat this solution in the same man-
ner as the sample.
Water <2.48> 23.3 – 26.3z (0.1 g, volumetric titration, solution as directed under Ultraviolet-visible Spectropho-
direct titration). tometry <2.24>.
Assay Weigh accurately about 0.2 g of Calcium Para- Dissolution rate (z) with respect to the labeled amount
aminosalicylate Hydrate, dissolve in 60 mL of water and of calcium paraaminosalicylate hydrate
0.75 mL of dilute hydrochloric acid by warming on a water (C7H5CaNO3.3 1/2 H2O)
bath. After cooling, add water to make exactly 100 mL, and ＝ MS/MT × AT/AS × 1/C × 900 × 1.330
use this solution as the sample solution. Measure exactly 30
MS: Amount (mg) of calcium paraaminosalicylate hydrate
mL of the sample solution, transfer to an iodine flask, and
for assay taken, calculated on the anhydrous basis
add exactly 25 mL of 0.05 mol/L bromine VS and 20 mL of
MT: Amount (g) of Calcium Paraaminosalicylate Gran-
a solution of potassium bromide (1 in 4). Add immediately
ules taken
14 mL of a mixture of acetic acid (100) and hydrochloric
C: Labeled amount (mg) of calcium paraaminosalicylate
acid (5:2), stopper the flask immediately, and allow to stand
hydrate (C7H5CaNO3.3 1/2 H2O) in 1 g
for 10 minutes with occasional shaking. Add cautiously 6
JP XVII Official Monographs / Anhydrous Dibasic Calcium Phosphate 557
Assay Powder Calcium Paraaminosalicylate Granules, protecting from light. Compare the opalescence developed in
weigh accurately a portion of the powder, equivalent to both solutions against a black background by viewing down-
about 0.2 g of calcium paraaminosalicylate hydrate ward or transversely. The opalescence developed in the test
(C7H5CaNO3.3 1/2 H2O), add 60 mL of water and 0.75 mL of solution is not more than that of the control solution. (not
dilute hydrochloric acid, and dissolve by heating on a water more than 0.25z)
bath. After cooling, add water to make exactly 100 mL, and (3) Sulfate—Dissolve 0.50 g of Anhydrous Dibasic Cal-
filter. Pipet 30 mL of the filtrate, transfer to an iodine flask, cium Phosphate in 5 mL of water and 5 mL of dilute hydro-
and proceed as directed in the Assay under Calcium Para- chloric acid, add water to make 100 mL, and filter, if neces-
aminosalicylate Hydrate. sary. Put 20 mL of this solution in a Nessler tube, add 1 mL
of dilute hydrochloric acid, and add water to make 50 mL,
Each mL of 0.05 mol/L bromine VS
and use this as the test solution. Transfer 1.0 mL of 0.005
＝ 4.238 mg of C7H5CaNO3.3 1/2 H2O
mol/L sulfuric acid VS to another Nessler tube, add 1 mL of
Containers and storage Containers—Tight containers. dilute hydrochloride acid and water to make 50 mL, and use
Storage—Light-resistant. this solution as the control solution. Add 2 mL of barium
chloride TS to the test solution and the control solution, mix
well, and allow to stand for 10 minutes. Compare the white
Anhydrous Dibasic Calcium turbidity produced in both solutions against a black back-
ground by viewing downward or transversely. The turbidity
Phosphate produced in the test solution is not thicker than that of the
control solution. (not more than 0.48z)
無水リン酸水素カルシウム
(4) Carbonate—Mix 1.0 g of Anhydrous Dibasic Cal-
cium Phosphate with 5 mL of freshly boiled and cooled
CaHPO4: 136.06 water, and add immediately 2 mL of hydrochloric acid: no
[7757-93-9] effervescence occurs.
(5) Heavy metals <1.07>—Dissolve 0.65 g of Anhydrous
This monograph is harmonized with the European Phar- Dibasic Calcium Phosphate in a mixture of 5 mL of water
macopoeia and the U.S. Pharmacopeia. The parts of the text and 5 mL of dilute hydrochloric acid by warming, cool, and
that are not harmonized are marked with symbols ( ). add ammonia TS until precipitates begin to form in the solu-
tion. Dissolve the precipitates by adding a small amount of
Anhydrous Dibasic Calcium Phosphate contains dilute hydrochloric acid dropwise, add 10 mL of hydrochlo-
not less than 98.0z and not more than 103.0z of ric acid-ammonium acetate buffer solution (pH 3.5) and
dibasic calcium phosphate (CaHPO4). water to make 50 mL, and perform the test using this solu-
Description tion as the test solution. Prepare the control solution as
Anhydrous Dibasic Calcium Phosphate oc-
follows: to 10 mL of hydrochloric acid-ammonium acetate
curs as white, crystalline powder or granules.
buffer solution (pH 3.5) add 2.0 mL of Standard Lead Solu-
tion and water to make 50 mL (not more than 31 ppm).
It dissolves in dilute hydrochloric acid and in dilute nitric
(6) Barium—Heat 0.5 g of Anhydrous Dibasic Calcium
acid.
Phosphate with 10 mL of water, add 1 mL of hydrochloric
Identification (1) Dissolve 0.1 g of Anhydrous Dibasic acid dropwise with stirring, and filter after cooling, if neces-
Calcium Phosphate in 10 mL of 2 mol/L hydrochloric acid sary. Add 2 mL of potassium sulfate TS to this solution, and
TS by warming, add 2.5 mL of ammonia TS dropwise with allow to stand for 10 minutes: no turbidity forms.
shaking, and add 5 mL of ammonium oxalate TS: a white (7) Arsenic <1.11>—Dissolve 1.0 g of Anhydrous Di-
precipitate is produced. basic Calcium Phosphate in 5 mL of dilute hydrochloric
(2) Dissolve 0.1 g of Anhydrous Dibasic Calcium Phos- acid, and perform the test with this solution as the test solu-
phate in 5 mL of dilute nitric acid, and add 2 mL of hexaam- tion (not more than 2 ppm).
monium heptamolybdate TS after warming at 709 C for 1 to
Loss on ignition <2.43> Not less than 6.6z and not more
2 minutes: a yellow precipitate is produced.
than 8.5z (1 g, 800 – 8259C, constant mass).
Purity (1) Acid-insoluble substances—Dissolve 5.0 g of
Assay Weigh accurately about 0.4 g of Anhydrous Dibasic
Anhydrous Dibasic Calcium Phosphate in 40 mL of water
Calcium Phosphate, dissolve in 12 mL of dilute hydrochloric
and 10 mL of hydrochloric acid, and boil gently for 5
acid by warming on a water bath, if necessary, and add
minutes. After cooling, collect the insoluble substance using
a filter paper for assay. Wash with water until no more tur-
add exactly 25 mL of 0.02 mol/L disodium dihydrogen
bidity of the washings is produced when silver nitrate TS is
ethylenediamine tetraacetate VS, 50 mL of water and 5 mL
added. Ignite to incinerate the residue and the filter paper at
of ammonia-ammonium chloride buffer solution (pH 10.7),
600 ± 509C: the mass is not more than 10 mg (not more
and titrate <2.50> the excess disodium dihydrogen ethylene-
than 0.2z).
diamine tetraacetate with 0.02 mol/L zinc sulfate VS (indica-
(2) Chloride—To 0.20 g of Anhydrous Dibasic Calcium
tor: 25 mg of eriochrome black T-sodium chloride indica-
Phosphate add 20 mL of water and 13 mL of dilute nitric
tor). Perform a blank determination in the same manner.
acid, dissolve by warming, if necessary, add water to make
100 mL, and filter, if necessary. Put 50 mL of this solution Each mL of 0.02 mol/L disodium dihydrogen
in a Nessler tube, and use this as the test solution. Transfer ethylenediamine tetraacetate VS
0.70 mL of 0.01 mol/L hydrochloric acid VS to another ＝ 2.721 mg of CaHPO4
Nessler tube, add 6 mL of dilute nitric acid and water to Containers and storage Containers—Well-closed contain-
make 50 mL, and use this solution as the control solution.
ers.
Add 1 mL of silver nitrate TS to the test solution and the
control solution, mix well, and allow to stand for 5 minutes
558 Dibasic Calcium Phosphate Hydrate / Official Monographs JP XVII
ground by viewing downward or transversely. The turbidity
Dibasic Calcium Phosphate Hydrate produced in the test solution is not thicker than that of the
control solution. (not more than 0.48z)
リン酸水素カルシウム水和物 (4) Carbonate—Mix 1.0 g of Dibasic Calcium Phosphate
Hydrate with 5 mL of freshly boiled and cooled water, and
add immediately 2 mL of hydrochloric acid: no efferves-
CaHPO4.2H2O: 172.09
cence occurs.
[7789-77-7] (5) Heavy metals <1.07>—Dissolve 0.65 g of Dibasic
Calcium Phosphate Hydrate in a mixture of 5 mL of water
and 5 mL of dilute hydrochloric acid by warming, cool, and
add ammonia TS until precipitates begin to form in the solu-
dilute hydrochloric acid dropwise, add 10 mL of hydrochlo-
Dibasic Calcium Phosphate Hydrate contains not
ric acid-ammonium acetate buffer solution (pH 3.5) and
less than 98.0z and not more than 105.0z of dibasic
water to make 50 mL, and perform the test using this solu-
calcium phosphate hydrate (CaHPO4.2H2O).
tion as the test solution. Prepare the control solution as

Description Dibasic Calcium Phosphate Hydrate occurs follows: to 10 mL of hydrochloric acid-ammonium acetate
as a white crystalline powder. buffer solution (pH 3.5) add 2.0 mL of Standard Lead Solu-
It is practically insoluble in water and in ethanol (99.5). tion and water to make 50 mL (not more than 31 ppm).
It dissolves in dilute hydrochloric acid and in dilute nitric (6) Barium—Heat 0.5 g of Dibasic Calcium Phosphate
acid. Hydrate with 10 mL of water, add 1 mL of hydrochloric acid
dropwise with stirring, and filter after cooling, if necessary.
Identification (1) Dissolve 0.1 g of Dibasic Calcium
Add 2 mL of potassium sulfate TS to this solution, and
Phosphate Hydrate in 10 mL of 2 mol/L hydrochloric acid
allow to stand for 10 minutes: no turbidity forms.
TS by warming, add 2.5 mL of ammonia TS dropwise with (7) Arsenic <1.11>—Dissolve 1.0 g of Dibasic Calcium
shaking, and add 5 mL of ammonium oxalate TS: a white
Phosphate Hydrate in 5 mL of dilute hydrochloric acid, and
precipitate is produced.
perform the test with this solution as the test solution (not
(2) Dissolve 0.1 g of Dibasic Calcium Phosphate Hy-
more than 2 ppm).
drate in 5 mL of dilute nitric acid, and add 2 mL of hexaam-
monium heptamolybdate TS after warming at 709 C for 1 to Loss on ignition <2.43> Not less than 24.5z and not more
2 minutes: a yellow precipitate is produced. than 26.5z (1 g, 800 – 8259
C, constant mass).
Purity (1) Acid-insoluble substance—Dissolve 5.0 g of Assay Weigh accurately about 0.4 g of Dibasic Calcium
Dibasic Calcium Phosphate Hydrate in 40 mL of water and Phosphate Hydrate, dissolve in 12 mL of dilute hydrochloric
10 mL of hydrochloric acid, and boil gently for 5 minutes. acid by warming on a water bath, if necessary, and add
After cooling, collect the insoluble substance using a filter water to make exactly 200 mL. Pipet 20 mL of this solution,
paper for assay. Wash with water until no more turbidity of add exactly 25 mL of 0.02 mol/L disodium dihydrogen
the washing is produced when silver nitrate TS is added. ethylenediamine tetraacetate VS, 50 mL of water and 5 mL
Ignite to incinerate the residue and filter paper at 600 ± of ammonia-ammonium chloride buffer solution (pH 10.7),
509 C: the mass is not more than 10 mg (not more than and titrate <2.50> the excess disodium dihydrogen ethylene-
0.2z). diamine tetraacetate with 0.02 mol/L zinc sulfate VS (indica-
(2) Chloride—To 0.20 g of Dibasic Calcium Phosphate tor: 25 mg of eriochrome black T-sodium chloride indica-
Hydrate add 20 mL of water and 13 mL of dilute nitric acid, tor). Perform a blank determination in the same manner.
dissolve by warming, if necessary, add water to make 100
mL, and filter, if necessary. Put 50 mL of this solution in a
Nessler tube, and use this as the test solution. Transfer 0.70
＝ 3.442 mg of CaHPO4.2H2O
mL of 0.01 mol/L hydrochloric acid VS to another Nessler
tube, add 6 mL of dilute nitric acid and water to make 50 Containers and storage Containers—Well-closed contain-
mL, and use this solution as the control solution. Add 1 mL ers.
of silver nitrate TS to the test solution and the control solu-
tion, mix well, and allow to stand for 5 minutes protecting
from light. Compare the opalescence developed in both solu- Monobasic Calcium Phosphate
tions against a black background by viewing downward or
transversely. The opalescence developed in the test solution Hydrate
is not more than that of the control solution. (not more than
リン酸二水素カルシウム水和物
0.25z)
(3) Sulfate—Dissolve 0.50 g of Dibasic Calcium Phos-
phate Hydrate in 5 mL of water and 5 mL of dilute hydro- Ca(H2PO4)2.H2O: 252.07
chloric acid, add water to make 100 mL, and filter, if neces-
sary. Put 20 mL of this solution in a Nessler tube, add 1 mL Monobasic Calcium Phosphate Hydrate, when
of dilute hydrochloric acid, and add water to make 50 mL, dried, contains not less than 90.0z of monobasic cal-
and use this as the test solution. Transfer 1.0 mL of 0.005 cium phosphate hydrate [Ca(H2PO4)2.H2O].
mol/L sulfuric acid VS to another Nessler tube, add 1 mL of
Description Monobasic Calcium Phosphate Hydrate oc-
dilute hydrochloride acid and water to make 50 mL, and use
curs as white, crystals or crystalline powder. It is odorless
this solution as the control solution. Add 2 mL of barium
and has an acid taste.
chloride TS to the test solution and the control solution, mix
well, and allow to stand for 10 minutes. Compare the white
turbidity produced in both solutions against a black back-
JP XVII Official Monographs / Calcium Polystyrene Sulfonate 559
It dissolves in dilute hydrochloric acid and in dilute nitric Each mL of 0.02 mol/L disodium dihydrogen
acid. ethylenediamine tetraacetate VS
It is slightly deliquescent. ＝ 5.041 mg of Ca(H2PO4)2.H2O
Identification (1) Dissolve 0.1 g of Monobasic Calcium Containers and storage Containers—Tight containers.
Phosphate Hydrate in 10 mL of diluted hydrochloric acid
(1 in 6) by warming, add 2.5 mL of ammonia TS dropwise
with shaking, and add 5 mL of ammonium oxalate TS: a Calcium Polystyrene Sulfonate
white precipitate is produced.
(2) Dissolve 0.1 g of Monobasic Calcium Phosphate Hy- ポリスチレンスルホン酸カルシウム
drate in 5 mL of dilute nitric acid, and add 2 mL of hexaam-
monium heptamolybdate TS after warming for 1 to 2
Calcium Polystyrene Sulfonate is a cation exchange
minutes at 709C: a yellow precipitate is produced.
resin prepared as the calcium form of the sulfonated
Purity (1) Clarity and color of solution—Dissolve 1.0 g styrene divinylbenzene copolymer.
of Monobasic Calcium Phosphate Hydrate in 19 mL of When dried, it contains not less than 7.0z and not
water and 2 mL of diluted hydrochloric acid (3 in 4), and more than 9.0z of calcium (Ca: 40.08).
heat on a water bath for 5 minutes with occasional shaking: Each g of Calcium Polystyrene Sulfonate, when
the solution is clear and colorless. dried, exchanges with 53 to 71 mg of potassium (K:
(2) Dibasic phosphate and acid—Triturate 1.0 g of 39.10).
Monobasic Calcium Phosphate Hydrate with 3 mL of water,
Description Calcium Polystyrene Sulfonate occurs as a
and add 100 mL of water and 1 drop of methyl orange TS: a
pale yellowish white to light yellow powder. It is odorless
red color develops. Then add 1.0 mL of 1 mol/L sodium hy-
and tasteless.
droxide VS: the color changes to yellow.
(3) Chloride <1.03>—Dissolve 1.0 g of Monobasic Cal-
diethyl ether.
cium Phosphate Hydrate in 20 mL of water and 12 mL of
dilute nitric acid, add water to make 100 mL, and filter, if Identification (1) Determine the infrared absorption spec-
necessary. Perform the test using 50 mL of this solution as trum of Calcium Polystyrene Sulfonate, previously dried, as
the test solution. Prepare the control solution with 0.25 mL directed in the potassium bromide disk method under In-
of 0.01 mol/L hydrochloric acid VS (not more than frared Spectrophotometry <2.25>, and compare the spectrum
0.018z). with the Reference Spectrum: both spectra exhibit similar in-
(4) Sulfate <1.14>—Dissolve 1.0 g of Monobasic Calcium tensities of absorption at the same wave numbers.
Phosphate Hydrate in 20 mL of water and 1 mL of hydro- (2) Mix 0.5 g of Calcium Polystyrene Sulfonate with 10
chloric acid, add water to make 100 mL, and filter, if neces- mL of dilute hydrochloric acid, filter, and neutralize the fil-
sary. Perform the test using 50 mL of this solution as the test trate with ammonia TS: the solution responds to the Qualita-
solution. Prepare the control solution with 0.50 mL of 0.005 tive Tests <1.09> for calcium salt.
Purity (1) Ammonium—Place 1.0 g of Calcium Polysty-
(5) Heavy metals <1.07>—Dissolve 0.65 g of Monobasic
rene Sulfonate in a flask, add 5 mL of sodium hydroxide TS,
Calcium Phosphate Hydrate in a mixture of 5 mL of water
cover the flask with a watch glass having a moistened strip of
and 5 mL of dilute hydrochloric acid by warming, cool, and
red litmus paper on the underside, and boil for 15 minutes:
add ammonia TS until precipitates begin to form in the solu-
the gas evolved does not change the red litmus paper to blue
dilute hydrochloric acid dropwise, add 10 mL of hydrochlo-
ric acid-ammonium acetate buffer solution (pH 3.5) and
Polystyrene Sulfonate according to Method 2, and perform
water to make 50 mL, and perform the test using this solu-
tion as the test solution. Prepare the control solution as
follows: to 10 mL of hydrochloric acid-ammonium acetate
buffer solution (pH 3.5) add 2.0 mL of Standard Lead Solu-
of Calcium Polystyrene Sulfonate according to Method 3,
tion and water to make 50 mL (not more than 31 ppm).
and perform the test (not more than 2 ppm).
(6) Arsenic <1.11>—Dissolve 1.0 g of Monobasic Cal-
(4) Styrene—To 10.0 g of Calcium Polystyrene Sulfonate
cium Phosphate Hydrate in 5 mL of dilute hydrochloric
add 10 mL of acetone, shake for 30 minutes, centrifuge, and
acid, and perform the test with this solution as the test solu-
tion (not more than 2 ppm).
dissolve 10 mg of styrene in acetone to make exactly 100 mL.
Loss on drying <2.41> Not more than 3.0z (1 g, silica gel, Pipet 1 mL of this solution, dilute with acetone to make
24 hours). exactly 100 mL, and use this solution as the standard solu-
Assay Weigh accurately about 0.4 g of Monobasic Calcium
solution and standard solution as directed under Gas Chro-
Phosphate Hydrate, previously dried, dissolve in 3 mL of
matography <2.02> according to the following conditions.
dilute hydrochloric acid, and add water to make exactly 100
Determine the peak heights, HT and HS, of styrene in each
mL. Pipet 20 mL of this solution, add exactly 25 mL of 0.02
solution: HT is not larger than HS.
mol/L disodium dihydrogen ethylenediamine tetraacetate
VS, 50 mL of water and 5 mL of ammonia-ammonium chlo-
ride buffer solution (pH 10.7), and titrate <2.50> the excess
disodium dihydrogen ethylenediamine tetraacetate with 0.02
and 2 m in length, having polyethylene glycol 20 M coated at
mol/L zinc acetate VS (indicator: 25 mg of eriochrome black
the ratio of 15z on siliceous earth for gas chromatography
T-sodium chloride indicator). Perform a blank determina-
(150 to 180 mm in particle diameter).
tion.
560 Calcium Polystyrene Sulfonate / Official Monographs JP XVII
909 C.
Carrier gas: Nitrogen.
Flow rate: Adjust so that the retention time of styrene is
about 9 minutes.
System performance: Mix 10 mg of styrene with 1000 mL
of acetone. When the procedure is run with 5 mL of this solu-
tion under the above operating conditions, the number of
theoretical plates and the symmetry factor of the peak of
styrene are not less than 800 and 0.8 to 1.2, respectively.
conditions, the relative standard deviation of the peak
heights of styrene is not more than 5z.
(5) Sodium—Pipet 2 mL of the 50-mL solution obtained
in the Assay (1), add 0.02 mol/L hydrochloric acid TS to
make exactly 500 mL, and use this solution as the sample so-
lution. Separately, weigh accurately 0.2542 g of sodium chlo-
ride, previously dried at 1309C for 2 hours, and dissolve in
0.02 mol/L hydrochloric acid TS to make exactly 1000 mL.
Pipet a suitable volume of this solution, and dilute with 0.02
mol/L hydrochloric acid TS to make a solution containing
1 to 3 mg of sodium (Na: 22.99) per mL, and use these solu-
tions as the standard solutions. Perform the test with the
sample solution and standard solutions according to the
Atomic Absorption Spectrophotometry <2.23> under the fol-
lowing conditions, and determine the amount of sodium in
the sample solution using the calibration curve obtained
from the standard solutions: the amount of sodium is not
more than 1z.
Supporting gas—Air.
Lamp: A sodium hollow-cathode lamp.
Loss on drying <2.41> Not more than 10.0z (1 g, in vacu- Fig. Andreasen pipet
um, 809C, 5 hours).
Microparticles (i) Apparatus: Use an apparatus as shown
in the illustration.
(ii) Procedure: Weigh accurately about 5.5 g of Calcium V: Actual volume (mL) to the mark of 20 cm at which the
Polystyrene Sulfonate, previously dried, add 300 mL of suction part of pipet is inserted
water of 259 C, and mix for 5 minutes. Transfer this turbid
Assay (1) Calcium—Weigh accurately about 1 g of Cal-
solution to the sedimentation tube J, keeping a temperature
cium Polystyrene Sulfonate, previously dried, and disperse
at 259C, add water of 259C to 2 mm below the mark F of 20
in 5 mL of 3 mol/L hydrochloric acid TS. Transfer this mix-
cm of the sedimentation tube J, and then insert the pipet.
ture, and wash out completely with the aid of a small quan-
Open the two-way stopcock C, exhaust air, add exactly
tity of 3 mol/L hydrochloric acid TS to a column 12 mm in
water from the vent-hole D to the mark F of 20 cm, and
inside diameter and 70 mm in length, packed with a pledged
close the two-way stopcock C. Shake the apparatus well ver-
of fine glass wool in the bottom of it, placing a 50-mL volu-
tically and horizontally, disperse Calcium Polystyrene Sul-
metric flask as a receiver under the column. Then collect
fonate in water, and then open the two-way stopcock, and
about 45 mL of eluate, adding 3 mol/L hydrochloric acid TS
allow to stand at 25 ± 19C for 5 hours and 15 minutes.
to the column, and add water to make exactly 50 mL. Pipet
Then, draw exactly the meniscus of the turbid solution in
20 mL of this solution, adjust with ammonia TS to a pH of
sedimentation tube J up to the mark of pipet bulb A by suc-
exactly 10. Titrate <2.50> immediately with 0.05 mol/L diso-
tion, open the two-way stopcock C to the outlet of pipet H,
dium dihydrogen ethylenediamine tetraacetate VS until the
and transfer exactly measured 20 mL of the turbid solution
red-purple color of the solution disappears, and a blue color
to a weighing bottle. Repeat the procedure, and combine
develops (indicator: 0.04 g eriochrome black T-sodium chlo-
exactly measured 20 mL of the turbid solution. Evaporate 20
ride indicator). Perform a blank determination, and make
mL of this turbid solution on a water bath to dryness, dry to
constant mass at 1059 C, and weigh the residue as MS (g).
Pipet 20 mL of used water, and weigh the residue in the same Each mL of 0.05 mol/L disodium dihydrogen
manner as MB (g). Calculate the difference mi (g) between ethylenediamine tetraacetate VS
MS and MB, and calculate the amount of microparticles (S ) ＝ 2.004 mg of Ca
by the following equation: the amount of microparticles is
(2) Potassium exchange capacity—Pipet 50 mL of Stand-
not more than 0.1z.
ard Potassium Stock Solution into a glass-stoppered flask
S (z) ＝ (mi × V )/(20 × MT) × 100 containing about 1 g of dried Calcium Polystyrene Sul-
fonate, accurately weighed, stir for 120 minutes, filter, and
MT: Amount (g) of Calcium Polystyrene Sulfonate taken
JP XVII Official Monographs / Calcium Sodium Edetate Hydrate 561
discard the first 20 mL of the filtrate. Pipet 5 mL of the sub- lution (28) (7 in 50), and add 3 mL of ammonium oxalate
sequent filtrate, and add 0.02 mol/L hydrochloric acid TS to TS: a white precipitate is formed.
make exactly 100 mL. Pipet 10 mL of this solution, add 0.02 (2) Determine the infrared absorption spectrum of Cal-
mol/L hydrochloric acid TS to make exactly 1000 mL, and cium Sodium Edetate Hydrate as directed in the potassium
use this solution as the sample solution. Separately, measure bromide disk method under Infrared Spectrophotometry
exactly a suitable volume of Standard Potassium Stock Solu- <2.25>, and compare the spectrum with the Reference Spec-
tion, dilute with 0.02 mol/L hydrochloric acid TS to make trum: both spectra exhibit similar intensities of absorption at
solutions containing 0.5 to 2.5 mg of potassium (K: 39.10) the same wave numbers.
per mL, and use these solutions as the standard solutions. (3) A solution of Calcium Sodium Edetate Hydrate (1 in
Perform the test with the sample solution and standard solu- 20) responds to the Qualitative Tests <1.09> (2) for sodium
tions as directed under Atomic Absorption Spectrophotome- salt.
try <2.23> according to the following conditions, and deter-
pH <2.54> The pH of a solution of 2.0 g of Calcium So-
mine the amount, Y (mg), of potassium in 1000 mL of the
dium Edetate Hydrate in 10 mL of water is 6.5 to 8.0.
sample solution, using the calibration curve obtained from
the standard solutions. The exchange quantity for potassium Purity (1) Clarity and color of solution—Dissolve
per g of dried Calcium Polystyrene Sulfonate is 53 to 71 mg, 0.25 g of Calcium Sodium Edetate Hydrate in 10 mL of
calculating by the following equation. water: the solution is clear and colorless.
(2) Chloride <1.03>—Dissolve 0.70 g of Calcium Sodium
Exchange quantity (mg) for potassium (K) per g of
Edetate Hydrate in water to make 20 mL. To this solution
dried Calcium Polystyrene Sulfonate
add 30 mL of dilute nitric acid, allow to stand for 30
＝ (X － 100 Y )/M
minutes, and filter. To 10 mL of the filtrate add water to
X: The amount (mg) of potassium in 50 mL of Standard make 50 mL, and perform the test using this solution as the
Potassium Stock Solution before exchange test solution. Prepare the control solution with 0.40 mL of
M: The amount (g) of dried Calcium Polystyrene Sul- 0.01 mol/L hydrochloric acid VS (not more than 0.10z).
fonate taken (3) Heavy metals <1.07>—Proceed with 1.0 g of Cal-
cium Sodium Edetate Hydrate according to Method 2, and
Supporting gas—Air.
Standard Lead Solution (not more than 20 ppm).
Lamp: A potassium hollow-cathode lamp.
(4) Disodium edetate—Dissolve 1.00 g of Calcium So-
dium Edetate Hydrate in 50 mL of water, add 5 mL of am-
Containers and storage Containers—Tight containers. monia-ammonium chloride buffer solution (pH 10.7), and
titrate <2.50> with 0.01 mol/L magnesium chloride VS until
the color of the solution changes from blue to red-purple (in-
Calcium Sodium Edetate Hydrate dicator: 40 mg of eriochrome black T-sodium chloride indi-
cator): the amount of 0.01 mol/L magnesium chloride VS
エデト酸カルシウムナトリウム水和物 consumed is not more than 3.0 mL (not more than 1.0z).

(5) Nitrilotriacetic acid—Conduct this procedure using
light-resistant vessels. Dissolve 0.100 g of Calcium Sodium
Edetate Hydrate in diluting solution to make exactly 25 mL,
and use this solution as the sample solution. Separately, dis-
solve 40.0 mg of nitrilotriacetic acid in diluting solution to
make exactly 100 mL. Pipet 1 mL of this solution, add 0.1
C10H12CaN2Na2O8.xH2O mL of the sample solution, then add diluting solution to
Disodium [{N, N?-ethane-1,2-diylbis[N- make exactly 100 mL, and use this solution as the standard
(carboxymethyl)glycinato]}(4-)-N, N?,O,O?,ON,ON?]calci- solution. Perform the test with exactly 20 mL each of the
ate(2-) hydrate sample solution and standard solution as directed under Liq-
[23411-34-9] uid Chromatography <2.01> according to the following con-
ditions, and determine the peak areas, AT and AS, of
This monograph is harmonized with the European Phar- nitrilotriacetic acid in each solution: AT is not larger than AS
macopoeia and the U.S. Pharmacopeia. The parts of the text (not more than 0.1z).
that are not harmonized are marked with symbols ( ). Diluting solution: Dissolve 10.0 g of iron (III) sulfate n-
hydrate in 20 mL of 0.5 mol/L sulfuric acid TS and 780 mL
Calcium Sodium Edetate Hydrate contains not less of water, adjust to pH 2.0 with sodium hydroxide TS, and
than 98.0z and not more than 102.0z of calcium dis- add water to make 1000 mL.
odium edetate (C10H12CaN2Na2O8: 374.27), calculated Operating conditions—
on the anhydrous basis. Detector: An ultraviolet absorption photometer (wave-
Description length: 273 nm).
Calcium Sodium Edetate Hydrate occurs as
white, powder or particles.
ter and 10 cm in length, packed with graphite carbon for liq-
uid chromatography (mean pore size: 25 nm, specific sur-
and practically insoluble in ethanol (99.5).
face: 120 m2/g, 5 mm in particle diameter).
It is hygroscopic.
Identification (1) Dissolve 2 g of Calcium Sodium 409C.
Edetate Hydrate in 10 mL of water, add 6 mL of a solution Mobile phase: Dissolve 50.0 mg of iron (III) sulfate n-
of lead (II) nitrate (33 in 1000), shake, and add 3 mL of po- hydrate in 50 mL of 0.5 mol/L sulfuric acid TS, add 750 mL
tassium iodide TS: no yellow precipitate is formed. Make of water, adjust to pH 1.5 with 0.5 mol/L sulfuric acid TS or
this solution alkaline by the addition of diluted ammonia so- sodium hydroxide TS, and add 20 mL of ethylene glycol and
562 Calcium Stearate / Official Monographs JP XVII
water to make 1000 mL. perform the test using this solution as the test solution. Pre-
Flow rate: 1.0 mL per minute (the retention time of pare the control solution by evaporating 2 mL of hydrochlo-
nitrilotriacetic acid is about 5 minutes). ric acid on a water bath to dryness and by adding 2 mL of
System suitability— dilute acetic acid, 2.0 mL of Standard Lead Solution and
Test for required detectability: When perform the test with water to make 50 mL (not more than 20 ppm).
20 mL of the standard solution under the above operating (2) Arsenic <1.11>—To 1.0 g of Calcium Stearate add 5
conditions, the SN ratio of the peak of nitrilotriacetic acid is mL of diluted hydrochloric acid (1 in 2) and 20 mL of chlo-
not less than 50. roform, shake vigorously for 3 minutes, allow to stand, and
System performance: When the procedure is run with 20 separate the water layer. Perform the test with the water
mL of the standard solution under the above operating con- layer as the test solution (not more than 2 ppm).
ditions, nitrilotriacetic acid and edetic acid are eluted in this
3 hours).
than 7.
System repeatability: When the test is repeated 6 times Assay Weigh accurately about 0.5 g of Calcium Stearate,
with 20 mL of the standard solution under the above operat- previously dried, heat gently with caution at first, and then
ing conditions, the relative standard deviation of the peak ignite gradually to ash. Cool, add 10 mL of dilute hydro-
area of nitrilotriacetic acid is not more than 1.0z. chloric acid to the residue, warm for 10 minutes on a water
bath, and transfer the contents to a flask with the aid of
10-mL, 10-mL, and 5-mL portions of hot water. Add so-
direct titration).
dium hydroxide TS until the solution becomes slightly
Assay Weigh accurately about 0.5 g of Calcium Sodium turbid, and then add 25 mL of 0.05 mol/L disodium dihy-
Edetate Hydrate, and dissolve in water to make exactly 200 drogen ethylenediamine tetraacetate VS, 10 mL of ammonia-
mL. Pipet 20 mL of this solution, add 80 mL of water, ammonium chloride buffer solution (pH 10.7), 4 drops of
adjust to pH 2 – 3 with dilute nitric acid, and titrate <2.50> eriochrome black T TS and 5 drops of methyl yellow TS,
with 0.01 mol/L bismuth nitrate VS until the color of the so- and titrate <2.50> rapidly the excess disodium dihydrogen
lution changes from yellow to red (indicator: 2 drops of ethylenediamine tetraacetate with 0.05 mol/L magnesium
xylenol orange TS). chloride VS, until the green color of the solution disappears
and a red color develops. Perform a blank determination.
Each mL of 0.01 mol/L bismuth nitrate VS
＝ 3.743 mg of C10H12CaN2Na2O8 Each mL of 0.05 mol/L disodium dihydrogen
Containers ethylenediamine tetraacetate VS
and storage Containers—Tight containers.
＝ 2.004 mg of Ca
Calcium Stearate ers.
ステアリン酸カルシウム
Camostat Mesilate
Calcium Stearate mainly consists of calcium salts カモスタットメシル酸塩
of stearic acid (C18H36O2: 284.48) and palmitic acid
(C16H32O2: 256.42).
Calcium Stearate, when dried, contains not less than
6.4z and not more than 7.1z of calcium (Ca: 40.08).
Description Calcium Stearate occurs as a white, light,
bulky powder. It feels smooth when touched, and is adhesive
to the skin. It is odorless or has a faint, characteristic odor.
It is practically insoluble in water, in ethanol (95) and in C20H22N4O5.CH4O3S: 494.52
diethyl ether. Dimethylcarbamoylmethyl
4-(4-guanidinobenzoyloxy)phenylacetate
Identification (1) Shake vigorously 3 g of Calcium
monomethanesulfonate
Stearate with 20 mL of diluted hydrochloric acid (1 in 2) and
[59721-29-8]
30 mL of diethyl ether for 3 minutes, and allow to stand: the
separated aqueous layer responds to the Qualitative Tests
<1.09> (1), (2) and (4) for calcium salt.
Camostat Mesilate, when dried, contains not less
than 98.5z of camostat mesilate (C20H22N4O5.
(2) Wash the diethyl ether layer obtained in (1) with 20
CH4O3S).
mL and 10 mL of dilute hydrochloric acid and 20 mL of
water successively, and evaporate the diethyl ether on a Description Camostat Mesilate occurs as white, crystals or
water bath: the residue melts <1.13> at a temperature not crystalline powder.
below 549 C. It is sparingly soluble in water, slightly soluble in ethanol
(95), and practically insoluble in diethyl ether.
Purity (1) Heavy metals <1.07>—Heat gently 1.0 g of
Calcium Stearate with caution at the beginning, and heat Identification (1) To 4 mL of a solution of Camostat
further, gradually raising the temperature, to incineration. Mesilate (1 in 2000) add 2 mL of 1-naphthol TS and 1 mL of
After cooling, add 2 mL of hydrochloric acid, evaporate on diacetyl TS, and allow to stand for 10 minutes: a red color
a water bath to dryness, warm the residue with 20 mL of develops.
water and 2 mL of dilute acetic acid for 2 minutes, cool, (2) Determine the absorption spectrum of a solution of
filter, and wash the residue with 15 mL of water. Combine Camostat Mesilate (1 in 100,000) as directed under Ultravio-
the filtrate and the washings, add water to make 50 mL, and let-visible Spectrophotometry <2.24>, and compare the spec-
JP XVII Official Monographs / d-Camphor 563
trum with the Reference Spectrum or the spectrum of a solu- Flow rate: Adjust so that the retention time of camostat is
tion of Camostat Mesilate RS prepared in the same manner about 10 minutes.
as the sample solution: both spectra exhibit similar intensi- System suitability—
ties of absorption at the same wavelengths. System performance: When the procedure is run with 2 mL
(3) A 0.1 g portion of Camostat Mesilate responds to the of the standard solution under the above operating condi-
Qualitative Tests <1.09> (1) for mesilate. tions, camostat and the internal standard are eluted in this
than 5.
Purity (1) Heavy metals <1.07>—Dissolve 1.0 g of System repeatability: When the test is repeated 6 times
Camostat Mesilate in 40 mL of water by warming, and add 2 with 2 mL of the standard solution under the above operating
mL of dilute acetic acid and water to make 50 mL. Perform conditions, the relative standard deviation of the ratio of the
the test using this solution as the test solution. Prepare the peak area of camostat to that of the internal standard is not
control solution with 2.0 mL of Standard Lead Solution and more than 1.0z.
2 mL of dilute acetic acid (not more than 20 ppm).
(2) Arsenic <1.11>—Dissolve 2.0 g of Camostat Mesilate
in 20 mL of 2 mol/L hydrochloric acid TS by heating in a
water bath, and continue to heat for 20 minutes. After cool-
ing, centrifuge, take 10 mL of the supernatant liquid, and d-Camphor
use this solution as the test solution. Perform the test (not
d-カンフル
more than 2 ppm).
(3) Related substances—Dissolve 30 mg of Camostat
Mesilate in 10 mL of ethanol (95), and use this solution as
ethanol (95) to make exactly 200 mL, and use this solution as
C10H16O: 152.23
(1R,4R)-1,7,7-Trimethylbicyclo[2.2.1]heptan-2-one
[464-49-3]
the plate with a mixture of ethyl acetate, water and acetic
acid (100) (3:1:1) to a distance of about 10 cm, and air-dry
d-Camphor contains not less than 96.0z of d-cam-
the plate. Allow the plate to stand overnight in iodine vapor:
phor (C10H16O).
the spots other than the principal spot from the sample solu-
tion are not more intense than the spot from the standard so- Description d-Camphor occurs as colorless or white, trans-
lution. lucent crystals, crystalline powder or masses. It has a charac-
teristic, agreeable odor, and a slightly bitter taste, followed
by a pleasant, cooling sensation.
1059C, 3 hours).
It is freely soluble in ethanol (95), in diethyl ether and in
Residue on ignition <2.44> Not more than 0.2z (1 g). carbon disulfide, and slightly soluble in water.
It slowly volatilizes at room temperature.
Assay Weigh accurately about 50 mg each of Camostat
Mesilate and Camostat Mesilate RS, previously dried, and Identification Dissolve 0.1 g of d-Camphor in 2 mL of
dissolve each in water to make exactly 50 mL. Pipet 5 mL methanol, add 1 mL of 2,4-dinitrophenylhydradine TS, and
each of these solutions, add exactly 5 mL of the internal heat for 5 minutes on a water bath: an orange-red precipitate
standard solution, and use these solutions as the sample so- is formed.
lution and the standard solution, respectively. Perform the
D : ＋41.0 – ＋43.09(5 g, ethanol
test with 2 mL each of the sample solution and standard solu-
(95), 50 mL, 100 mm).
cording to the following conditions, and calculate the ratios, Melting point <2.60> 177 – 1829C
QT and QS, of the peak area of camostat to that of the inter-
Purity (1) Water—Shake 1.0 g of d-Camphor with 10 mL
nal standard.
of carbon disulfide: the solution is clear.
Amount (mg) of camostat mesilate (C20H22N4O5.CH4O3S) (2) Chlorinated compounds—Mix 0.20 g of finely pow-
＝ M S × Q T / QS dered d-Camphor with 0.4 g of sodium peroxide in a dried
porcelain crucible. Heat the crucible gently by the open
MS: Amount (mg) of Camostat Mesilate RS taken
flame until the incineration is complete. Dissolve the residue
Internal standard solution—A solution of butyl parahy- in 20 mL of warm water, acidify with 12 mL of dilute nitric
droxybenzoate in ethanol (95) (1 in 1500). acid, and filter the solution into a Nessler tube. Wash the
Operating conditions— filter paper with three 5-mL portions of hot water, adding
Detector: An ultraviolet absorption photometer (wave- the washings to the filtrate. After cooling, add water to
length: 265 nm). make 50 mL, then add 1 mL of silver nitrate TS, mix well,
Column: A stainless steel column 4.6 mm in inside diame- and allow to stand for 5 minutes: the turbidity of the solu-
ter and 15 cm in length, packed with octadecylsilanized silica tion does not exceed that of the following control solution.
gel for liquid chromatography (5 mm in particle diameter). Control solution: Prepare in the same manner as described
Column temperature: A constant temperature of about above, using 0.20 mL of 0.01 mol/L hydrochloric acid VS.
259 C. (3) Non-volatile residue—Heat 2.0 g of d-Camphor on a
Mobile phase: A mixture of methanol, a solution of so- water bath until sublimation is complete, then dry the
dium 1-heptane sulfonate (1 in 500), a solution of sodium residue at 1059C for 3 hours: the mass of the residue does
lauryl sulfate (1 in 1000) and acetic acid (100) (200:100:50:1). not exceed 1.0 mg.
564 dl-Camphor / Official Monographs JP XVII
Assay Weigh accurately about 0.1 g each of d-Camphor Optical rotation <2.49> [a]20
D : －1.5 – ＋1.59(5 g, ethanol
and d-Camphor RS, add exactly 5 mL each of the internal (95), 50 mL, 100 mm).
standard solution, dissolve in ethanol (99.5) to make 100
Melting point <2.60> 175 – 1809
C
mL, and use these solutions as the sample solution and the
standard solution. Perform the test with 2 mL each of the Purity (1) Water—Shake 1.0 g of dl-Camphor with 10
sample solution and standard solution as directed under Gas mL of carbon disulfide: the solution is clear.
Chromatography <2.02> according to the following condi- (2) Chlorinated compounds—Mix 0.20 g of finely pow-
tions, and calculate the ratios, QT and QS, of the peak area dered dl-Camphor with 0.4 g of sodium peroxide in a dried
of d-camphor to that of the internal standard. porcelain crucible. Heat the crucible gently by the open
flame until the incineration is complete. Dissolve the residue
Amount (mg) of d-camphor (C10H16O) ＝ MS × QT/QS
in 20 mL of warm water, acidify with 12 mL of dilute nitric
MS: Amount (mg) of d-Camphor RS taken acid, and filter the solution into a Nessler tube. Wash the
filter paper with three 5-mL portions of hot water, adding
Internal standard solution—A solution of methyl salicylate
the washings to the filtrate. After cooling, add water to
in ethanol (99.5) (1 in 25).
make 50 mL, then add 1 mL of silver nitrate TS, mix well,
and allow to stand for 5 minutes: the turbidity of the solu-
tion does not exceed that of the following control solution.
Column: A glass column 3 mm in inside diameter and 3 m
Control solution: Prepare in the same manner as described
in length, which is packed with 10z of polyethylene glycol
above, using 0.20 mL of 0.01 mol/L hydrochloric acid VS.
20 M for gas chromatography supported on 180 to 250 mm
(3) Non-volatile residue—Heat 2.0 g of dl-Camphor on a
mesh silanized siliceous earth for gas chromatography.
water bath until sublimation is complete, then dry the
residue at 1059C for 3 hours: the mass of the residue does
1609C.
not exceed 1.0 mg.
Flow rate: Adjust so that the retention time of d-camphor Assay Weigh accurately about 0.1 g each of dl-Camphor
is about 6 minutes. and dl-Camphor RS, add exactly 5 mL each of the internal
System suitability— standard solution, dissolve in ethanol (99.5) to make 100
System performance: When the procedure is run with 2 mL mL, and use these solutions as the sample solution and
of the standard solution under the above operating condi- standard solution, respectively. Perform the test with 2 mL
tions, d-camphor and the internal standard are eluted in this each of the sample solution and standard solution as directed
order with the resolution between these peaks being not less under Gas Chromatography <2.02> according to the follow-
than 7. ing conditions, and calculate the ratios, QT and QS, of the
System repeatability: When the test is repeated 6 times peak area of dl-camphor to that of the internal standard.
Amount (mg) of dl-camphor (C10H16O) ＝ MS × QT/QS
conditions, the relative standard deviation of the ratios of
the peak area of d-camphor to that of the internal standard MS: Amount (mg) of dl-Camphor RS taken
Internal standard solution—A solution of methyl salicylate
Containers and storage Containers—Tight containers. in ethanol (99.5) (1 in 25).
dl-Camphor Column: A glass column 3 mm in inside diameter and 3 m
in length, which is packed with 10z of polyethylene glycol
dl-カンフル 20 M for gas chromatography supported on 180 to 250 mm
mesh silanized siliceous earth for gas chromatography.
1609C.
Flow rate: Adjust so that the retention time of dl-camphor
is about 6 minutes.
C10H16O: 152.23 System suitability—
(1RS,4RS )-1,7,7-Trimethylbicyclo[2.2.1]heptan-2-one System performance: When the procedure is run with 2 mL
[76-22-2] of the standard solution under the above operating condi-
tions, dl-camphor and the internal standard are eluted in this
dl-Camphor contains not less than 96.0z of dl-cam- order with the resolution between these peaks being not less
phor (C10H16O). than 7.
Description dl-Camphor occurs as colorless or white,
translucent crystals, crystalline powder or masses. It has a
characteristic, agreeable odor, and has a slightly bitter taste
the peak area of dl-camphor to that of the internal standard
followed by a pleasant, cooling sensation.
It is freely soluble in ethanol (95), in diethyl ether and in
carbon disulfide, and slightly soluble in water. Containers and storage Containers—Tight containers.
It slowly volatilizes at room temperature.
Identification Dissolve 0.1 g of dl-Camphor in 2 mL of
methanol, add 1 mL of 2,4-dinitrophenylhydradine TS, and
heat for 5 minutes on a water bath: an orange-red precipitate
is formed.
JP XVII Official Monographs / Candesartan Cilexetil 565
times the peak area of candesartan cilexetil from the stand-

Candesartan Cilexetil ard solution, the area of the peak other than candesartan
cilexetil and the peaks mentioned above from the sample so-
カンデサルタンシレキセチル lution is not smaller than 1/10 times the peak area of can-
desartan cilexetil from the standard solution, and the total
area of the peaks other than candesartan cilexetil from the
candesartan cilexetil from the standard solution.
length: 254 nm).
C33H34N6O6: 610.66
(1RS )-1-(Cyclohexyloxycarbonyloxy)ethyl-2-ethoxy-
259C.
1-{[2?-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl}-
Mobile phase A: A mixture of acetonitrile, water and
1H-benzimidazole-7-carboxylate
acetic acid (100) (57:43:1).
[145040-37-5]
Mobile phase B: A mixture of acetonitrile, water and
acetic acid (100) (90:10:1).
Candesartan Cilexetil contains not less than 99.0z
and not more than 101.0z of candesartan cilexetil
(C33H34N6O6), calculated on the anhydrous basis.
Description Candesartan Cilexetil occurs as white crystals Time after injection Mobile phase A Mobile phase B
or a white crystalline powder. of sample (min) (volz) (volz)
It is soluble in acetic acid (100), sparingly soluble in meth-
anol, slightly soluble in ethanol (99.5), and practically in- 0 – 30 100 → 0 0 → 100
soluble in water.
A solution of Candesartan Cilexetil in methanol (1 in 100) Flow rate: 0.8 mL per minute.
shows no optical rotation. Time span of measurement: For 30 minutes after injec-
Candesartan Cilexetil shows crystal polymorphism. tion, beginning after the solvent peak.
Identification (1) Determine the absorption spectrum of a System suitability—
solution of Candesartan Cilexetil in methanol (1 in 50,000) Test for required detectability: Pipet 2 mL of the standard
as directed under Ultraviolet-visible Spectrophotometry solution, and add a mixture of acetonitrile and water (3:2) to
<2.24>, and compare the spectrum with the Reference Spec- make exactly 20 mL. Confirm that the peak area of can-
trum: both spectra exhibit similar intensities of absorption at desartan cilexetil obtained with 10 mL of this solution is
the same wavelengths. equivalent to 7 to 13z of that obtained with 10 mL of the
(2) Determine the infrared absorption spectrum of Can- standard solution.
desartan Cilexetil as directed in the potassium bromide disk System performance: When the procedure is run with 10
method under Infrared Spectrophotometry <2.25>, and com- mL of the standard solution under the above operating con-
pare the spectrum with the Reference Spectrum: both spectra ditions, the number of theoretical plates and the symmetry
exhibit similar intensities of absorption at the same wave factor of the peak of candesartan cilexetil are not less than
numbers. If any difference appears between the spectra, 12,000 and not more than 1.5, respectively.
recrystallize the sample and the RS according to the method System repeatability: When the test is repeated 6 times
otherwise specified, filter and dry the crystals, and perform with 10 mL of the standard solution under the above operat-
the test with the crystals. ing conditions, the relative standard deviation of the peak
area of candesartan cilexetil is not more than 2.0z.
Candesartan Cilexetil according to Method 4, and perform Water <2.48> Not more than 0.3z (0.5 g, coulometric
the test. Prepare the control solution with 2.0 mL of Stand- titration).
ard Lead Solution (not more than 20 ppm). Residue on ignition <2.44> Not more than 0.1z (1 g).
(2) Related substances—Dissolve 20 mg of Candesartan
Cilexetil in 50 mL of a mixture of acetonitrile and water Assay Weigh accurately about 0.5 g of Candesartan Cilex-
(3:2), and use this solution as the sample solution. Pipet 1 etil, dissolve in 60 mL of acetic acid (100), and titrate <2.50>
mL of the sample solution, add a mixture of acetonitrile and with 0.1 mol/L perchloric acid VS (potentiometric titration).
water (3:2) to make exactly 100 mL, and use this solution as Perform a blank determination in the same manner, and
the standard solution. Perform the test with exactly 10 mL make any necessary correction.
each of the sample solution and standard solution as directed Each mL of 0.1 mol/L perchloric acid VS
under Liquid Chromatography <2.01> according to the fol- ＝ 61.07 mg of C33H34N6O6
tomatic integration method: the area of the peaks, having Containers and storage Containers—Well-closed contain-
the relative retention time of about 0.4 and about 2.0 to can- ers.
desartan cilexetil, obtained from the sample solution is not
larger than 1/5 times the peak area of candesartan cilexetil
obtained from the standard solution, the area of the peak,
having the relative retention time of about 0.5 to candesar-
tan cilexetil, from the sample solution is not larger than 3/10
566 Candesartan Cilexetil Tablets / Official Monographs JP XVII
Candesartan Cilexetil Tablets Time after injection

of sample (min)
Mobile phase A
(volz)
Mobile phase B
(volz)
カンデサルタンシレキセチル錠
0 – 30 100 → 0 0 → 100
Candesartan Cilexetil Tablets contain not less than Flow rate: 0.8 mL per minute.
95.0z and not more than 105.0z of the labeled Time span of measurement: For 30 minutes after injec-
amount of candesartan cilexetil (C33H34N6O6: 610.66). tion, beginning after the solvent peak.
Method of preparation Prepare as directed under Tablets, System suitability—
with Candesartan Cilexetil. Test for required detectability: Pipet 2 mL of the standard
solution, and add a mixture of acetonitrile and water (3:2) to
Identification Powder Candesartan Cilexetil Tablets. To a make exactly 20 mL. Confirm that the peak area of can-
portion of the powder, equivalent to 1 mg of Candesartan desartan cilexetil obtained with 10 mL of this solution is
Cilexetil, add 50 mL of methanol, shake vigorously for 10 equivalent to 7 to 13z of that obtained with 10 mL of the
minutes, and filter. Determine the absorption spectrum of standard solution.
the filtrate as directed under Ultraviolet-visible Spectropho- System performance: When the procedure is run with 10
tometry <2.24>: it exhibits absorption maxima between 252 mL of the standard solution under the above operating con-
nm and 256 nm and between 302 nm and 307 nm. ditions, the number of theoretical plates and the symmetry
Purity Related substances—Powder not less than 10 Can- factor of the peak of candesartan cilexetil are not less than
desartan Cilexetil Tablets. To a portion of the powder, 12,000 and not more than 1.5, respectively.
equivalent to 6 mg of Candesartan Cilexetil, add 15 mL of a System repeatability: When the test is repeated 6 times
mixture of acetonitrile and water (3:2), shake vigorously for with 10 mL of the standard solution under the above operat-
10 minutes, and centrifuge. Filter the supernatant liquid ing conditions, the relative standard deviation of the peak
through a membrane filter with a pore size not exceeding area of candesartan cilexetil is not more than 2.0z.
0.45 mm. Discard the first 3 mL of the filtrate, and use the Uniformity of dosage units <6.02> Perform the test accord-
subsequent filtrate as the sample solution. Pipet 1 mL of the ing to the following method: it meets the requirement of the
sample solution, add a mixture of acetonitrile and water Content uniformity test.
(3:2) to make exactly 100 mL, and use this solution as the To 1 tablet of Candesartan Cilexetil Tablets add 30 mL of
standard solution. Perform the test with exactly 10 mL each a mixture of acetonitrile and water (3:2), shake vigorously
of the sample solution and standard solution as directed for 20 minutes, then add a mixture of acetonitrile and water
under Liquid Chromatography <2.01> according to the fol- (3:2) to make exactly V mL so that each mL contains about
lowing conditions, and determine each peak area by the au- 40 mg of candesartan cilexetil (C33H34N6O6), centrifuge, and
tomatic integration method: the area of the peak having the use the supernatant liquid as the sample solution. Separately,
relative retention time of about 0.5 to candesartan cilexetil weigh accurately about 50 mg of candesartan cilexetil for
obtained from the sample solution is not larger than 1.5 assay (separately determine the water <2.48> in the same
times the peak area of candesartan cilexetil obtained from manner as Candesartan Cilexetil), and dissolve in aceto-
the standard solution, the area of the peak having the rela- nitrile to make exactly 50 mL. Pipet 4 mL of this solution,
tive retention time of about 0.8, about 1.1 and about 1.5 to add a mixture of acetonitrile and water (3:2) to make exactly
candesartan cilexetil from the sample solution is not larger 100 mL, and use this solution as the standard solution. De-
than 1/2 times the peak area of candesartan cilexetil from termine the absorbances, AT and AS, of the sample solution
the standard solution, the area of the peak having the rela- and standard solution at 305 nm as directed under Ultravio-
tive retention time of about 2.0 to candesartan cilexetil from let-visible Spectrophotometry <2.24>.
the sample solution is not larger than the peak area of can-
desartan cilexetil from the standard solution, the area of the Amount (mg) of candesartan cilexetil (C33H34N6O6)
peak other than candesartan cilexetil, the peak having the ＝ MS × AT/AS × V/1250
relative retention time of about 0.4 to candesartan cilexetil MS: Amount (mg) of candesartan cilexetil for assay taken,
and the peaks mentioned above from the sample solution is calculated on the anhydrous basis
smaller than 1/10 times the peak area of candesartan cilexetil
from the standard solution, and the total area of the peaks Dissolution <6.10> When the test is performed at 50 revolu-
other than candesartan cilexetil from the sample solution is tions per minute according to the Paddle method, using 900
not larger than 4 times the peak area of candesartan cilexetil mL of a solution of polysorbate 20 (1 in 100) as the dissolu-
from the standard solution. tion medium, the dissolution rate in 45 minutes of Candesar-
Operating conditions— tan Cilexetil Tablets is not less than 75z.
Detector: An ultraviolet absorption photometer (wave- Start the test with 1 tablet of Candesartan Cilexetil
length: 254 nm). Tablets, withdraw not less than 20 mL of the medium at the
Column: A stainless steel column 3.9 mm in inside diame- specified minute after starting the test, and filter through a
ter and 15 cm in length, packed with octadecylsilanized silica membrane filter with a pore size not exceeding 0.45 mm. Dis-
gel for liquid chromatography (4 mm in particle diameter). card the first 5 mL of the filtrate, pipet V mL of the subse-
Column temperature: A constant temperature of about quent filtrate, add the dissolution medium to make exactly
259 C. V? mL so that each mL contains about 2.2 mg of candesartan
Mobile phase A: A mixture of acetonitrile, water and cilexetil (C33H34N6O6), and use this solution as the sample so-
acetic acid (100) (57:43:1). lution. Separately, weigh accurately about 50 mg of can-
Mobile phase B: A mixture of acetonitrile, water and desartan cilexetil for assay (separately determine the water
acetic acid (100) (90:10:1). <2.48> in the same manner as Candesartan Cilexetil), and dis-
Flowing of mobile phase: Control the gradient by mixing solve in acetonitrile to make exactly 50 mL. Pipet 5 mL of
the mobile phases A and B as directed in the following table. this solution, add acetonitrile to make exactly 50 mL. Pipet 1
JP XVII Official Monographs / Candesartan Cilexetil and Amlodipine Besylate Tablets 567
mL of this solution, add the dissolution medium to make ex- acid (100) (57:43:1).
actly 50 mL, and use this solution as the standard solution. Flow rate: Adjust so that the retention time of candesar-
Perform the test with exactly 50 mL each of the sample solu- tan cilexetil is about 13 minutes.
tion and standard solution as directed under Liquid Chroma- System suitability—
tography <2.01>, and determine the peak areas, AT and AS, System performance: When the procedure is run with 10
of candesartan cilexetil in each solution. mL of the standard solution under the above operating con-
ditions, the internal standard and candesartan cilexetil are
of candesartan cilexetil (C33H34N6O6)
＝ MS × AT/AS × V?/V × 1/C × 18/5
MS: Amount (mg) of candesartan cilexetil for assay taken, with 10 mL of the standard solution under the above operat-
calculated on the anhydrous basis ing conditions, the relative standard deviation of the ratio of
C: Labeled amount (mg) of candesartan cilexetil the peak area of candesartan cilexetil to that of the internal
(C33H34N6O6) in 1 tablet standard is not more than 1.0z.
Operating conditions— Containers and storage Containers—Tight containers.
Assay.
System suitability— Candesartan Cilexetil and
mL of the standard solution under the above operating con- Amlodipine Besylate Tablets
カンデサルタンシレキセチル･アムロジピンベシル酸塩錠
factor of the peak of candesartan cilexetil are not less than
7000 and not more than 1.5, respectively.
System repeatability: When the test is repeated 6 times Candesartan Cilexetil and Amlodipine Besylate
with 50 mL of the standard solution under the above operat- Tablets contain not less than 95.0z and not more
ing conditions, the relative standard deviation of the peak than 105.0z of the labeled amount of candesartan
area of candesartan cilexetil is not more than 2.0z. cilexetil (C33H34N6O6: 610.66) and amlodipine besylate
(C20H25ClN2O5.C6H6O3S: 567.05).
Assay Weigh accurately the mass of not less than 20 Can-
desartan Cilexetil Tablets, and powder. Weigh accurately a Method of preparation Prepare as directed under Tablets,
portion of the powder, equivalent to about 6 mg of can- with Candesartan Cilexetil and Amlodipine Besylate.
desartan cilexetil (C33H34N6O6), add exactly 15 mL of the in-
Identification (1) Shake thoroughly a quantity of pow-
ternal standard solution, then add a mixture of acetonitrile
dered Candesartan Cilexetil and Amlodipine Besylate
and water (3:2) to make 150 mL, shake vigorously for 10
Tablets, equivalent to 8 mg of Candesartan Cilexetil, with 20
minutes, and allow to stand. Filter the supernatant liquid
mL of 0.01 mol/L hydrochloric acid TS, and centrifuge.
Remove the supernatant liquid, to the residue add 20 mL of
0.45 mm. Discard the first 5 mL of the filtrate, and use the
0.01 mol/L hydrochloric acid TS, shake thoroughly, and
centrifuge. Remove the supernatant liquid, to the residue
accurately about 50 mg of candesartan cilexetil for assay
add 40 mL of methanol, shake thoroughly, and filter
Candesartan Cilexetil), dissolve in acetonitrile to make ex-
0.45 mm. To 5 mL of the filtrate add methanol to make 50
of the internal standard solution, then add a mixture of
acetonitrile and water (3:2) to make 100 mL, and use this so-
<2.24>: it exhibits maxima between 252 nm and 256 nm, and
lution as the standard solution. Perform the test with 10 mL
(2) Shake thoroughly a quantity of powdered Candesar-
tan Cilexetil and Amlodipine Besylate Tablets, equivalent to
2.5 mg of Amlodipine Besylate, with 20 mL of 0.01 mol/L
the peak area of candesartan cilexetil to that of the internal
hydrochloric acid TS, and centrifuge. Filter the supernatant
standard.
liquid through a membrane filter with a pore size not
Amount (mg) of candesartan cilexetil (C33H34N6O6) exceeding 0.45 mm. To 5 mL of the filtrate add methanol to
＝ MS × QT/QS × 3/25 make 25 mL, and determine the absorption spectrum of this
MS: Amount (mg) of candesartan cilexetil for assay taken,
tometry <2.24>: it exhibits maxima between 236 nm and 240
calculated on the anhydrous basis
nm, and between 360 nm and 364 nm.
Internal standard solution—A solution of acenaphthene in
Purity Related substances—Shake vigorously for 20
acetonitrile (1 in 800).
minutes a quantity of powdered Candesartan Cilexetil and
Amlodipine Besylate Tablets, equivalent to 8 mg of Can-
desartan Cilexetil, with 20 mL of diluting solution, and filter
length: 254 nm).
this solution through a membrane filter with a pore size not
exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and
use the subsequent filtrate as the sample solution. Pipet 1
mL of the sample solution, add diluting solution to make ex-
259 C.
Mobile phase: A mixture of acetonitrile, water and acetic
568 Candesartan Cilexetil and Amlodipine Besylate Tablets / Official Monographs JP XVII
tography <2.01> according to the following conditions. De- (1) Candesartan cilexetil—To 1 tablet of Candesartan
termine each peak area by the automatic integration method: Cilexetil and Amlodipine Besylate Tablets add exactly 20 mL
the area of the peak, having the relative retention time of of diluting solution, shake for 20 minutes to disintegrate the
about 0.8 to candesartan cilexetil, obtained from the sample tablet, and filter through a membrane filter with a pore size
solution is not larger than 1.5 times the peak area of can- not exceeding 0.45 mm. Discard the first 5 mL of the filtrate,
desartan cilexetil obtained from the standard solution, the pipet V mL of the subsequent filtrate, add exactly V?/5 mL
area of the peaks, having a relative retention time of about of the internal standard solution, then add diluting solution
0.9, about 1.1 and about 1.2 from the sample solution is not to make V? mL so that each mL contains about 0.16 mg of
larger than 1/2 times the peak area of candesartan cilexetil candesartan cilexetil (C33H34N6O6), and use this solution as
from the standard solution, the area of the peak, having a the sample solution. Then, proceed as directed in the Assay
relative retention time of about 1.4 from the sample solu- (1).
tion, is not larger than the peak area of candesartan cilexetil
Amount (mg) of candesartan cilexetil (C33H34N6O6)
from the standard solution, and the area of the peak other
＝ MS × QT/QS × V?/V × 2/25
than candesartan cilexetil and the peaks mentioned above
from the sample solution is smaller than 1/10 times the peak MS: Amount (mg) of candesartan cilexetil for assay taken,
area of candesartan cilexetil from the standard solution. Fur- calculated on the anhydrous basis
thermore, the total area of the peaks other than candesartan
cilexetil from the sample solution is not larger than 4 times
droxybenzoate in diluting solution (1 in 2500).
the peak area of candesartan cilexetil from the standard so-
Diluting solution: To 3.5 mL of triethylamine add water
lution.
to make 500 mL, and adjust to pH 3.0 with phosphoric acid.
Diluting solution: To 3.5 mL of triethylamine add water
To 400 mL of this solution add 600 mL of acetonitrile.
to make 500 mL, and adjust to pH 3.0 with phosphoric acid.
(2) Amlodipine besylate—To 1 tablet of Candesartan
Cilexetil and Amlodipine Besylate Tablets add exactly 20 mL
of diluting solution, shake for 20 minutes to disintegrate the
tablet, and filter through a membrane filter with a pore size
length: 253 nm).
not exceeding 0.45 mm. Discard the first 5 mL of the filtrate,
pipet V mL of the subsequent filtrate, add exactly V?/5 mL
of the internal standard solution, then add diluting solution
to make V? mL so that each mL contains about 70 mg of
amlodipine besylate (C20H25ClN2O5.C6H6O3S), and use this
259 C.
solution as the sample solution. Then, proceed as directed in
Mobile phase A: A mixture of water, acetonitrile and
the Assay (2).
trifluoroacetic acid (4000:1000:1).
Mobile phase B: A mixture of acetonitrile, water and Amount (mg) of amlodipine besylate
trifluoroacetic acid (4000:1000:1). (C20H25ClN2O5.C6H6O3S)
Flowing of mobile phase: Control the gradient by mixing ＝ MS × QT/QS × V?/V × 1/25
MS: Amount (mg) of Amlodipine Besylate RS taken, cal-
Time after injection Mobile phase A Mobile phase B
of sample (min) (volz) (volz) Internal standard solution—A solution of butyl parahy-
droxybenzoate in diluting solution (1 in 2500).
0 – 15 100 → 50 0 → 50 Diluting solution: To 3.5 mL of triethylamine add water
15 – 50 50 → 0 50 → 100 to make 500 mL, and adjust to pH 3.0 with phosphoric acid.
50 – 60 0 100 To 400 mL of this solution add 600 mL of acetonitrile.
Dissolution <6.10> (1) Candesartan cilexetil—When the
Flow rate: 1.0 mL per minute. test is performed at 75 revolutions per minute according to
Time span of measurement: For 60 minutes after injec- the Paddle method, using 900 mL of a solution, prepared by
tion, beginning after the solvent peak. dissolving 1 g of polysorbate 80 in 2nd fluid for dissolution
System suitability— test to make 1000 mL, as the dissolution medium, the disso-
Test for required detectability: To exactly 1 mL of the lution rate in 45 minutes of Candesartan Cilexetil and
standard solution add diluting solution to make exactly 50 Amlodipine Besylate Tablets is not less than 80z.
mL. Confirm that the peak area of candesartan cilexetil ob- Start the test with 1 tablet of Candesartan Cilexetil and
tained with 20 mL of this solution is equivalent to 1.4 to Amlodipine Besylate Tablets, withdraw not less than 10 mL
2.6z of that obtained with 20 mL of the standard solution. of the medium at the specified minute after starting the test,
System performance: When the procedure is run with 20 and filter through a membrane filter with a pore size not
mL of the standard solution under the above operating con- exceeding 0.45 mm. Discard the first 5 mL of the filtrate,
ditions, the number of theoretical plates and the symmetry pipet V mL of the subsequent filtrate, add the dissolution
factor of the peak of candesartan cilexetil are not less than medium to make exactly V? mL so that each mL contains
100,000 and not more than 1.5, respectively. about 8.9 mg of candesartan cilexetil (C33H34N6O6), and use
System repeatability: When the test is repeated 6 times this solution as the sample solution. Separately, weigh
with 20 mL of the standard solution under the above operat- accurately about 45 mg of candesartan cilexetil for assay
ing conditions, the relative standard deviation of the peak (separately, determine the water <2.48> in the same manner
area of candesartan cilexetil is not more than 2.0z. as Candesartan Cilexetil), and dissolve in acetonitrile to
Uniformity of dosage units <6.02> Perform the test accord- make exactly 50 mL. Pipet 1 mL of this solution, add the
ing to the following methods: it meets the requirements of dissolution medium to make exactly 100 mL, and use this so-
the Content uniformity test. lution as the standard solution. Perform the test with exactly
JP XVII Official Monographs / Candesartan Cilexetil and Amlodipine Besylate Tablets 569
20 mL each of the sample solution and standard solution as C: Labeled amount (mg) of amlodipine besylate
directed under Liquid Chromatography <2.01> according to (C20H25ClN2O5.C6H6O3S) in 1 tablet
the following conditions, and determine the peak areas, AT
and AS, of candesartan cilexetil in each solution.
Detector, column, column temperature, and mobile phase:
Dissolution rate (z) with respect to the labeled amount Proceed as directed in the operating conditions in the Assay
of candesartan cilexetil (C33H34N6O6) (1).
＝ MS × AT/AS × V?/V × 1/C × 18 Flow rate: Adjust so that the retention time of amlodipine
is about 4 minutes.
C: Labeled amount (mg) of candesartan cilexetil
(C33H34N6O6) in 1 tablet
Operating conditions— factor of the peak of amlodipine are not less than 3000 and
Detector: An ultraviolet absorption photometer (wave- not more than 2.0, respectively.
length: 254 nm). System repeatability: When the test is repeated 6 times
Column: A stainless steel column 4.6 mm in inside diame- with 50 mL of the standard solution under the above operat-
ter and 5 cm in length, packed with octadecylsilanized silica ing conditions, the relative standard deviation of the peak
gel for liquid chromatography (5 mm in particle diameter). area of amlodipine is not more than 1.0z.
Assay (1) Candesartan cilexetil—Weigh accurately the
259 C.
mass of not less than 20 Candesartan Cilexetil and Amlodi-
pine Besylate Tablets, and powder. Weigh accurately a por-
acid (100) (57:43:1).
tion of the powder, equivalent to about 8 mg of candesartan
Flow rate: Adjust so that the retention time of candesar-
cilexetil (C33H34N6O6), add exactly 20 mL of diluting solu-
tan cilexetil is about 6.5 minutes.
tion, shake vigorously for 20 minutes, and filter through a
quent filtrate, add exactly 5 mL of the internal standard
solution, then add diluting solution to make 25 mL, and use
this solution as the sample solution. Separately, weigh
2000 and not more than 1.5, respectively.
(separately, determine the water <2.48> in the same manner
as Candesartan Cilexetil), dissolve in diluting solution to
make exactly 100 mL, and use this solution as the candesar-
tan cilexetil standard stock solution. Pipet 10 mL of the can-
(2) Amlodipine besylate—When the test is performed at
desartan cilexetil standard stock solution, add exactly 5 mL
50 revolutions per minute according to the Paddle method,
of the internal standard solution, then add diluting solution
using 900 mL of 0.05 mol/L acetic acid-sodium acetate
to make 25 mL, and use this solution as the standard solu-
buffer solution (pH 4.0) as the dissolution medium, the dis-
tion. Perform the test with 10 mL each of the sample solution
solution rate in 30 minutes of Candesartan Cilexetil and
Amlodipine Besylate Tablets is not less than 80z.
raphy <2.01> according to the following conditions, and cal-
Start the test with 1 tablet of Candesartan Cilexetil and
culate the ratios, QT and QS, of the peak area of candesartan
Amlodipine Besylate Tablets, withdraw not less than 10 mL
cilexetil to that of the internal standard.
of the medium at the specified minute after starting the test,
and filter through a membrane filter with a pore size not ex- Amount (mg) of candesartan cilexetil (C33H34N6O6)
ceeding 0.45 mm. Discard the first 5 mL of the filtrate, pipet ＝ MS × QT/QS × 1/5
V mL of the subsequent filtrate, add the dissolution medium
to make exactly V? mL so that each mL contains about 3.9
mg of amlodipine besylate (C20H25ClN2O5.C6H6O3S), and use
this solution as the sample solution. Separately, weigh accu- Internal standard solution—A solution of butyl parahy-
rately about 39 mg of Amlodipine Besylate RS (separately, droxybenzoate in diluting solution (1 in 2500).
determine the water <2.48> in the same manner as Amlodi- Diluting solution: To 3.5 mL of triethylamine add water
pine Besylate), and dissolve in acetonitrile to make exactly 50 to make 500 mL, and adjust to pH 3.0 with phosphoric acid.
mL. Pipet 5 mL of this solution, add acetonitrile to make ex- To 400 mL of this solution add 600 mL of acetonitrile.
actly 50 mL. Pipet 5 mL of this solution, add the dissolution Operating conditions—
medium to make exactly 100 mL, and use this solution as the Detector: An ultraviolet absorption photometer (wave-
standard solution. Perform the test with exactly 50 mL each length: 238 nm).
of the sample solution and standard solution as directed Column: A stainless steel column 3.9 mm in inside diame-
under Liquid Chromatography <2.01> according to the fol- ter and 15 cm in length, packed with octadecylsilanized silica
lowing conditions, and determine the peak areas, AT and AS, gel for liquid chromatography (5 mm in particle diameter).
of amlodipine in each solution. Column temperature: A constant temperature of about
259C.
Mobile phase: To 7 mL of triethylamine add water to
of amlodipine besylate (C20H25ClN2O5.C6H6O3S)
make 1000 mL, and adjust to pH 6.5 with phosphoric acid.
＝ MS × AT/AS × V?/V × 1/C × 9
MS: Amount (mg) of Amlodipine Besylate RS taken, cal- Flow rate: Adjust so that the retention time of candesar-
culated on the anhydrous basis tan cilexetil is about 31 minutes.
570 Candesartan Cilexetil and Hydrochlorothiazide Tablets / Official Monographs JP XVII
System suitability— amlodipine and the internal standard is not less than 15.
System performance: Mix 10 mL of the candesartan cilex- System repeatability: When the test is repeated 6 times
etil standard stock solution and 5 mL of the amlodipine be- with 10 mL of the standard solution under the above operat-
sylate standard stock solution prepared in the Assay (2), add ing conditions, the relative standard deviation of the ratio of
5 mL of the internal standard solution, then add diluting so- the peak area of amlodipine to that of the internal standard
lution to make 25 mL. When the procedure is run with 10 mL is not more than 1.0z.
of this solution under the above operating conditions, am-
lodipine, the internal standard and candesartan cilexetil are
eluted in this order and the resolution between the peaks of
the internal standard and candesartan cilexetil is not less
than 15. Candesartan Cilexetil and
System repeatability: When the test is repeated 6 times Hydrochlorothiazide Tablets
ing conditions, the relative standard deviation of the ratio of カンデサルタンシレキセチル･ヒドロクロロチアジド錠
Candesartan Cilexetil and Hydrochlorothiazide
(2) Amlodipine besylate—Weigh accurately the mass of
Tablets contain not less than 95.0z and not more
not less than 20 Candesartan Cilexetil and Amlodipine Besy-
than 105.0z of the labeled amount of candesartan
late Tablets, and powder. Weigh accurately a portion of the
cilexetil (C33H34N6O6: 610.66) and hydrochlorothiazide
powder, equivalent to about 3.5 mg of amlodipine besylate
(C7H8ClN3O4S2: 297.74).
(C20H25ClN2O5.C6H6O3S), add exactly 20 mL of diluting so-
lution, shake vigorously for 20 minutes, and filter through a Method of preparation Prepare as directed under Tablets,
membrane filter with a pore size not exceeding 0.45 mm. Dis- with Candesartan Cilexetil and Hydrochlorothiazide.
Identification (1) To an amount of powdered Candesar-
tan Cilexetil and Hydrochlorothiazide Tablets, equivalent to
lution, then add diluting solution to make 25 mL, and use
4 mg of Candesartan Cilexetil, add 5 mL of acetone, shake
thoroughly, centrifuge, and filter the supernatant liquid
rately about 35 mg of Amlodipine Besylate RS (separately,
determine the water <2.48> in the same manner as Amlodi-
0.45 mm. Evaporate the filtrate to dryness with the aid of a
pine Besylate), dissolve in diluting solution to make exactly
current of nitrogen. Dissolve the residue in 0.5 mL of ace-
100 mL, and use this solution as the amlodipine besylate
tone, and use this solution as the sample solution. Sepa-
standard stock solution. Pipet 5 mL of the amlodipine besy-
rately, dissolve 40 mg of candesartan cilexetil in 5 mL of ace-
late standard stock solution, add exactly 5 mL of the internal
tone, and use this solution as the standard solution. Perform
standard solution, then add diluting solution to make 25
the test with these solutions as directed under Thin-layer
Chromatography <2.03>. Spot 2 mL each of the sample solu-
the test with 10 mL each of the sample solution and standard
tion and standard solution on a plate of silica gel with
fluorescent indicator for thin-layer chromatography. De-
velop the plate with a mixture of ethyl acetate and acetic acid
ratios, QT and QS, of the peak area of amlodipine to that of
(100) (10:1) to a distance of about 10 cm, and air-dry the
Amount (mg) of amlodipine besylate nm): the R f value of the spot having a larger R f value among
(C20H25ClN2O5.C6H6O3S) the spots obtained from the sample solution is the same with
＝ MS × QT/QS × 1/10 that of the spot obtained from the standard solution.
(2) To an amount of powdered Candesartan Cilexetil
MS: Amount (mg) of Amlodipine Besylate RS taken, cal-
and Hydrochlorothiazide Tablets, equivalent to 6.25 mg of
Hydrochlorothiazide, add 5 mL of acetone, shake thor-
Internal standard solution—A solution of butyl parahy- oughly, centrifuge, and filter the supernatant liquid through
droxybenzoate in diluting solution (1 in 2500). a membrane filter with a pore size not exceeding 0.45 mm.
Diluting solution: To 3.5 mL of triethylamine add water Evaporate the filtrate to dryness with the aid of a current of
to make 500 mL, and adjust to pH 3.0 with phosphoric acid. nitrogen. Dissolve the residue in 0.5 mL of acetone, and use
To 400 mL of this solution add 600 mL of acetonitrile. this solution as the sample solution. Separately, dissolve 50
Operating conditions— mg of hydrochlorothiazide in 4 mL of acetone, and use this
Detector, column, column temperature, and mobile phase: solution as the standard solution. Perform the test with these
Proceed as directed in the operating conditions in the Assay solutions as directed under Thin-layer Chromatography
(1). <2.03>. Spot 2 mL each of the sample solution and standard
Flow rate: Adjust so that the retention time of amlodipine solution on a plate of silica gel with fluorescent indicator for
is about 2.5 minutes. thin-layer chromatography. Develop the plate with a mixture
System suitability— of ethyl acetate and acetic acid (100) (10:1) to a distance of
System performance: Mix 10 mL of the candesartan cilex- about 10 cm, and air-dry the plate. Examine under ultravio-
etil standard stock solution prepared in the Assay (1) and 5 let light (main wavelength: 254 nm): the R f value of the spot
mL of the amlodipine besylate standard stock solution, add having a smaller R f value among the spots obtained from the
5 mL of the internal standard solution, then add diluting so- sample solution is the same with that of the spot obtained
lution to make 25 mL. When the procedure is run with 10 mL from the standard solution.
of this solution under the above operating conditions, am-
Purity Related substances—(i) To an amount of powdered
lodipine, the internal standard and candesartan cilexetil are
Candesartan Cilexetil and Hydrochlorothiazide Tablets,
eluted in this order and the resolution between the peaks of
equivalent to 4 mg of Candesartan Cilexetil, add 10 mL of a
JP XVII Official Monographs / Candesartan Cilexetil and Hydrochlorothiazide Tablets 571
mixture of acetonitrile and water (3:2), shake vigorously for relative retention time of about 0.9 and about 3.2 to hydro-
10 minutes, centrifuge, and filter the supernatant liquid chlorothiazide, obtained from the sample solution is not
through a membrane filter with a pore size not exceeding larger than the peak area of hydrochlorothiazide obtained
0.45 mm. Discard the first 5 mL of the filtrate, and use the from the standard solution, and the area of the peak, other
subsequent filtrate as the sample solution. Pipet 1 mL of the than hydrochlorothiazide and the peaks mentioned above,
sample solution, add a mixture of acetonitrile and water from the sample solution is not larger than 1/5 times the
(3:2) to make exactly 100 mL, and use this solution as the peak area of hydrochlorothiazide from the standard solu-
standard solution. Perform the test with exactly 10 mL each tion. Furthermore, the total area of the peaks other than hy-
of the sample solution and standard solution as directed drochlorothiazide from the sample solution is not larger than
under Liquid Chromatography <2.01> according to the fol- 2 times the peak area of hydrochlorothiazide from the stand-
lowing conditions, and determine each peak area by the au- ard solution. For the area of the peak, having a relative
tomatic integration method: the area of the peak, having a retention time of about 0.8 and about 0.9 to hydrochloro-
relative retention time of about 0.5 to candesartan cilexetil, thiazide, multiply their relative response factors, 1.4 and 0.5,
obtained from the sample solution is not larger than 1.5 respectively.
times the peak area of candesartan cilexetil obtained from Operating conditions—
the standard solution, the area of the peak, having a relative Detector, column, column temperature, mobile phase, and
retention time of about 0.8, about 1.1 and about 1.5, from flow rate: Proceed as directed in the operating conditions in
the sample solution is not larger than 1/2 times the peak area the Assay (2).
of candesartan cilexetil from the standard solution, the area Time span of measurement: For 30 minutes after injec-
of the peak, having a relative retention time of about 2.0, tion, beginning after the solvent peak.
from the sample solution is not larger than the peak area of System suitability—
candesartan cilexetil from the standard solution, and the Test for required detectability: To exactly 1 mL of the
area of the peak, other than candesartan cilexetil and the standard solution add a mixture of 0.05 mol/L sodium dihy-
peaks mentioned above, from the sample solution is smaller drogen phosphate TS (pH 3.0) and acetonitrile (3:1) to make
than 1/10 times the peak area of candesartan cilexetil from exactly 50 mL. Confirm that the peak area of hydrochloro-
the standard solution. Furthermore, the total area of the thiazide obtained with 10 mL of this solution is equivalent to
peaks other than candesartan cilexetil from the sample solu- 1.4z to 2.6z of that obtained with 10 mL of the standard
tion is not larger than 4 times the peak area of candesartan solution.
cilexetil from the standard solution. System performance: When the procedure is run with 10
Operating conditions— mL of the standard solution under the above operating con-
Detector, column, column temperature, mobile phases A ditions, the number of theoretical plates and the symmetry
and B, flowing of mobile phase, and time span of meas- factor of the peak of hydrochlorothiazide are not less than
urement: Proceed as directed in the operating conditions in 5000 and not more than 1.5, respectively.
the Purity (2) under Candesartan Cilexetil. System repeatability: When the test is repeated 6 times
Flow rate: 0.6 mL per minute. with 10 mL of the standard solution under the above operat-
System suitability— ing conditions, the relative standard deviation of the peak
Test for required detectability: To exactly 1 mL of the area of hydrochlorothiazide is not more than 2.0z.
ing to the following methods: it meets the requirements of
candesartan cilexetil obtained with 10 mL of this solution is
the Content uniformity test.
equivalent to 1.4z to 2.6z of that obtained with 10 mL of
(1) Candesartan cilexetil—To 1 tablet of Candesartan
Cilexetil and Hydrochlorothiazide Tablets add exactly V/10
mL of the internal standard solution, add a mixture of aceto-
nitrile and 0.05 mol/L sodium dihydrogen phosphate TS
(pH 3.0) (3:2) to make V mL so that each mL contains about
40 mg of candesartan cilexetil (C33H34N6O6). Shake for 20
12,000 and not more than 1.5, respectively.
minutes to disintegrate the tablet, centrifuge, and use the su-
pernatant liquid as the sample solution. Separately, weigh
(ii) To an amount of powdered Candesartan Cilexetil
actly 50 mL, and use this solution as the candesartan cilexetil
and Hydrochlorothiazide Tablets, equivalent to 6.25 mg of
standard stock solution. Pipet 4 mL of the candesartan cilex-
Hydrochlorothiazide, add 10 mL of a mixture of 0.05 mol/L
etil standard stock solution, add exactly 10 mL of the inter-
sodium dihydrogen phosphate TS (pH 3.0) and acetonitrile
nal standard solution, add a mixture of acetonitrile and 0.05
(3:1), shake vigorously for 10 minutes, centrifuge, and filter
mol/L sodium dihydrogen phosphate TS (pH 3.0) (3:2) to
the supernatant liquid through a membrane filter with a pore
trate, and use the subsequent filtrate as the sample solution.
Pipet 1 mL of the sample solution, add a mixture of 0.05
mol/L sodium dihydrogen phosphate TS (pH 3.0) and aceto-
the ratios, QT and QS, of the peak area of candesartan cilex-
nitrile (3:1) to make exactly 100 mL, and use this solution as
etil to that of the internal standard.
each of the sample solution and standard solution as directed Amount (mg) of candesartan cilexetil (C33H34N6O6)
under Liquid Chromatography <2.01> according to the fol- ＝ MS × QT/QS × V × 1/1250
tomatic integration method: the area of the peak, having a
calculated on the anhydrous basis.
572 Candesartan Cilexetil and Hydrochlorothiazide Tablets / Official Monographs JP XVII
Internal standard solution—A solution of benzophenone in sodium dihydrogen phosphate TS (pH 5.5) (11:9).
acetonitrile (1 in 10,000). Flow rate: Adjust so that the retention time of hydrochlo-
Operating conditions— rothiazide is about 3.5 minutes.
Detector: An ultraviolet absorption photometer (wave- System suitability—
length: 254 nm). System performance: Mix 4 mL of the candesartan cilex-
Column: A stainless steel column 4.6 mm in inside diame- etil standard stock solution obtained in (1) and 10 mL of the
ter and 15 cm in length, packed with octadecylsilanized silica hydrochlorothiazide standard stock solution, add 10 mL of
gel for liquid chromatography (4 mm in particle diameter). the internal standard solution, and add a mixture of aceto-
Column temperature: A constant temperature of about nitrile and 0.05 mol/L sodium dihydrogen phosphate TS
259 C. (pH 3.0) (3:2) to make 100 mL. When the procedure is run
Mobile phase: A mixture of acetonitrile and 0.05 mol/L with 10 mL of this solution under the above operating condi-
sodium dihydrogen phosphate TS (pH 5.5) (11:9). tions, hydrochlorothiazide, candesartan cilexetil and the in-
Flow rate: Adjust so that the retention time of candesar- ternal standard are eluted in this order, and the resolution
tan cilexetil is about 7 minutes. between the peaks of hydrochlorothiazide and candesartan
System suitability— cilexetil is not less than 7, and the resolution between the
System performance: Mix 4 mL of the candesartan cilex- peaks of candesartan cilexetil and the internal standard is
etil standard stock solution and 10 mL of the hydrochloro- not less than 6.
thiazide standard stock solution obtained in (2), add 10 mL System repeatability: When the test is repeated 6 times
of the internal standard solution, and add a mixture of with 10 mL of the standard solution under the above operat-
acetonitrile and 0.05 mol/L sodium dihydrogen phosphate ing conditions, the relative standard deviation of the ratio of
TS (pH 3.0) (3:2) to make 100 mL. When the procedure is the peak area of hydrochlorothiazide to that of the internal
run with 10 mL of this solution under the above operating standard is not more than 1.0z.
conditions, hydrochlorothiazide, candesartan cilexetil and
Dissolution <6.10> (1) Candesartan cilexetil—When the
the internal standard are eluted in this order, and the resolu-
test is performed at 50 revolutions per minute according to
tion between the peaks of hydrochlorothiazide and candesar-
the Paddle method, using 900 mL of a solution, prepared by
tan cilexetil is not less than 7, and the resolution between the
dissolving 1 g of polysorbate 80 in 2nd fluid for dissolution
peaks of candesartan cilexetil and the internal standard is
test to make 1000 mL, as the dissolution medium, the disso-
not less than 6.
lution rate in 45 minutes of Candesartan Cilexetil and Hy-
drochlorothiazide Tablets is not less than 75z.
Hydrochlorothiazide Tablets, withdraw not less than 10 mL
(2) Hydrochlorothiazide—To 1 tablet of Candesartan
Cilexetil and Hydrochlorothiazide Tablets add exactly V/10
pipet V mL of the subsequent filtrate, add 0.05 mol/L so-
mL of the internal standard solution, add a mixture of aceto-
dium dihydrogen phosphate TS (pH 3.0) to make exactly
nitrile and 0.05 mol/L sodium dihydrogen phosphate TS
V? mL so that each mL contains about 2.2 mg of candesartan
(pH 3.0) (3:2) to make V mL so that each mL contains about
cilexetil (C33H34N6O6), and use this solution as the sample
63 mg of hydrochlorothiazide (C7H8ClN3O4S2). Shake for 20
solution. Separately, weigh accurately about 25 mg of can-
minutes, centrifuge, and use the supernatant liquid as the
desartan cilexetil for assay (separately determine the water
<2.48> in the same manner as Candesartan Cilexetil), dissolve
of Hydrochlorothiazide RS (separately determine the loss on
in acetonitrile to make exactly 100 mL, and use this solution
drying <2.41> under the same conditions as Hydrochloro-
as the candesartan cilexetil standard stock solution. Pipet 2
thiazide), dissolve in acetonitrile to make exactly 50 mL, and
mL of the candesartan cilexetil standard stock solution, add
use this solution as the hydrochlorothiazide standard stock
dissolution medium to make exactly 100 mL. Pipet 10 mL of
solution. Pipet 10 mL of the hydrochlorothiazide standard
this solution, add 0.05 mol/L sodium dihydrogen phosphate
stock solution, add exactly 10 mL of the internal standard
TS (pH 3.0) to make exactly 20 mL, and use this solution as
solution, add a mixture of acetonitrile and 0.05 mol/L so-
dium dihydrogen phosphate TS (pH 3.0) (3:2) to make 100
lowing conditions, and determine the peak areas, AT and AS,
of candesartan cilexetil in each solution.
ratios, QT and QS, of the peak area of hydrochlorothiazide Dissolution rate (z) with respect to the labeled amount
to that of the internal standard. of candesartan cilexetil (C33H34N6O6)
＝ MS × AT/AS × V?/V × 1/C × 9
Amount (mg) of hydrochlorothiazide (C7H8ClN3O4S2)
＝ MS × QT/QS × V × 1/500 MS: Amount (mg) of candesartan cilexetil for assay taken,
MS: Amount (mg) of Hydrochlorothiazide RS taken, cal-
C: Labeled amount (mg) of candesartan cilexetil
(C33H34N6O6) in 1 tablet
Internal standard solution—A solution of benzophenone in
acetonitrile (1 in 10,000).
Uniformity of dosage units (1).
directed in the operating conditions in the Assay (2).
System performance: Mix 2 mL each of the candesartan
Mobile phase: A mixture of acetonitrile and 0.05 mol/L
cilexetil standard stock solution and the hydrochlorothiazide
JP XVII Official Monographs / Candesartan Cilexetil and Hydrochlorothiazide Tablets 573
standard stock solution obtained in (2), and add the dissolu- with the resolution between these peaks being not less than 6.
tion medium to make 100 mL. To 10 mL of this solution add System repeatability: When the test is repeated 6 times
10 mL of 0.05 mol/L sodium dihydrogen phosphate TS (pH with 40 mL of the standard solution under the above operat-
3.0). When the procedure is run with 40 mL of this solution ing conditions, the relative standard deviation of the peak
under the above operating conditions, hydrochlorothiazide area of hydrochlorothiazide is not more than 1.0z.
and candesartan cilexetil are eluted in this order with the
Assay (1) Candesartan cilexetil—Weigh accurately the
mass of not less than 20 Candesartan Cilexetil and Hydro-
chlorothiazide Tablets, and powder. Weigh accurately a por-
tion of the powder, equivalent to about 4 mg of candesartan
cilexetil (C33H34N6O6), add exactly 10 mL of the internal
standard solution, add a mixture of acetonitrile and water
(2) Hydrochlorothiazide—When the test is performed at
(3:2) to make 100 mL, and shake vigorously for 10 minutes.
50 revolutions per minute according to the Paddle method,
Allow to stand for 5 minutes, and filter the supernatant liq-
using 900 mL of a solution, prepared by dissolving 1 g of
uid through a membrane filter with a pore size not exceeding
polysorbate 80 in 2nd fluid for dissolution test to make 1000
0.45 mm. Discard the first 5 mL of the filtrate, and use the
mL, as the dissolution medium, the dissolution rate in 45
minutes of Candesartan Cilexetil and Hydrochlorothiazide
Hydrochlorothiazide Tablets, withdraw not less than 10 mL
of the internal standard solution, add a mixture of aceto-
nitrile and water (3:2) to make 100 mL, and use this solution
as the standard solution. Perform the test with 10 mL each of
pipet V mL of the subsequent filtrate, add 0.05 mol/L so-
dium dihydrogen phosphate TS (pH 3.0) to make exactly
V? mL so that each mL contains about 3.5 mg of hydrochlo-
rothiazide (C7H8ClN3O4S2), and use this solution as the sam-
area of candesartan cilexetil to that of the internal standard.
Hydrochlorothiazide RS (separately determine the loss on Amount (mg) of candesartan cilexetil (C33H34N6O6)
drying <2.41> under the same conditions as Hydrochloro- ＝ MS × QT/QS × 2/25
thiazide), dissolve in acetonitrile to make exactly 100 mL,
and use this solution as the hydrochlorothiazide standard
stock solution. Pipet 2 mL of the hydrochlorothiazide stand-
ard stock solution, add dissolution medium to make exactly Internal standard solution—A solution of acenaphthene in
100 mL. Pipet 10 mL of this solution, add 0.05 mol/L so- acetonitrile (1 in 800).
dium dihydrogen phosphate TS (pH 3.0) to make exactly 20 Operating conditions—
mL, and use this solution as the standard solution. Perform Detector: An ultraviolet absorption photometer (wave-
the test with exactly 40 mL each of the sample solution and length: 254 nm).
standard solution as directed under Liquid Chromatography Column: A stainless steel column 4 mm in inside diameter
<2.01> according to the following conditions, and determine and 15 cm in length, packed with octadecylsilanized silica gel
the peak areas, AT and AS, of hydrochlorothiazide in each for liquid chromatography (4 mm in particle diameter).
solution. Column temperature: A constant temperature of about
259C.
of hydrochlorothiazide (C7H8ClN3O4S2)
acid (100) (57:43:1).
＝ MS × AT/AS × V?/V × 1/C × 9
Flow rate: Adjust so that the retention time of candesar-
MS: Amount (mg) of Hydrochlorothiazide RS taken, cal- tan cilexetil is about 13 minutes.
culated on the dried basis System suitability—
C: Labeled amount (mg) of hydrochlorothiazide System performance: When the procedure is run with 10
(C7H8ClN3O4S2) in 1 tablet mL of the standard solution under the above operating con-
ditions, the internal standard and candesartan cilexetil are
directed in the operating conditions in the Assay (2).
Mobile phase: A mixture of acetonitrile and 0.05 mol/L
sodium dihydrogen phosphate TS (pH 5.5) (11:9).
Flow rate: Adjust so that the retention time of hydrochlo-
rothiazide is about 3.5 minutes.
(2) Hydrochlorothiazide—Weigh accurately the mass of
System performance: Mix 2 mL each of the candesartan
not less than 20 Candesartan Cilexetil and Hydrochloro-
cilexetil standard stock solution obtained in (1) and the hy-
thiazide Tablets, and powder. Weigh accurately a portion of
drochlorothiazide standard stock solution, and add the dis-
the powder, equivalent to about 6.25 mg of hydrochloro-
solution medium to make 100 mL. To 10 mL of this solution
thiazide (C7H8ClN3O4S2), add exactly 10 mL of the internal
add 10 mL of 0.05 mol/L sodium dihydrogen phosphate TS
standard solution, add a mixture of 0.05 mol/L sodium di-
(pH 3.0). When the procedure is run with 40 mL of this solu-
hydrogen phosphate TS (pH 3.0) and acetonitrile (3:1) to
tion under the above operating conditions, hydrochloro-
make 100 mL, and shake vigorously for 10 minutes. Allow
thiazide and candesartan cilexetil are eluted in this order
to stand for 5 minutes, and filter the supernatant liquid
574 Capsules / Official Monographs JP XVII
through a membrane filter with a pore size not exceeding conical flask, add 50 mL of water, and shake often, keeping
0.45 mm. Discard the first 5 mL of the filtrate, and use the the temperature at 37 ± 29C. Perform this test 5 times: they
subsequent filtrate as the sample solution. Separately, weigh all dissolve within 10 minutes. All these solutions are odor-
accurately about 31 mg of Hydrochlorothiazide RS less, and neutral or slightly acidic.
(separately determine the loss on drying <2.41> under the
Loss on drying <2.41> 13 – 16z (1 g, 1059
C, 2 hours).
same conditions as Hydrochlorothiazide), dissolve in aceto-
nitrile to make exactly 50 mL. Pipet 10 mL of this solution, Microbial limit <4.05> The acceptance criteria of TAMC
add exactly 10 mL of the internal standard solution, add a and TYMC are 103 CFU/g and 102 CFU/g, respectively.
mixture of 0.05 mol/L sodium dihydrogen phosphate TS
(pH 3.0) and acetonitrile (3:1) to make 100 mL, and use this
ers.
solution as the standard solution. Perform the test with 10
mL each of the sample solution and standard solution as
the following conditions, and calculate the ratios, QT and Hypromellose Capsules
QS, of the peak area of hydrochlorothiazide to that of the
ヒプロメロースカプセル
internal standard.
Amount (mg) of hydrochlorothiazide (C7H8ClN3O4S2)
＝ MS × QT/QS × 1/5
Hypromellose Capsules are made of Hypromellose
as the base material, and their shape is a pair of cylin-
MS: Amount (mg) of Hydrochlorothiazide RS taken, cal- ders with one end closed which can be overlapped on
culated on the dried basis each other.
The label states the use or nonuse of the gelling
Internal standard solution—A solution of m-hydroxya-
agent and its name.
cetophenone in acetonitrile (1 in 6500).
Operating conditions— Method of preparation Dissolve Hypromellose in water by
Detector: An ultraviolet absorption photometer (wave- warming, add, if necessary, Glycerin or D-Sorbitol, emulsifi-
length: 254 nm). ers, dispersing agents, preservatives, coloring agents, gelling
Column: A stainless steel column 4.6 mm in inside diame- agents, and gelling aid, etc. to make a viscous liquid, and
ter and 15 cm in length, packed with octadecylsilanized silica form into a certain shape while warming.
gel for liquid chromatography (4 mm in particle diameter). They may be coated with a lubricant as necessary.
Solubility and acidity or alkalinity Place one pair of
259 C.
Hypromellose Capsules without snapping in a 100-mL coni-
Mobile phase: A mixture of 0.05 mol/L sodium dihydro-
cal flask, add 50 mL of water, and shake occasionally at 37
gen phosphate TS (pH 5.5) and acetonitrile (3:1).
± 29 C. When perform this test 5 times, either capsule dis-
Flow rate: Adjust so that the retention time of hydrochlo-
solves within 15 minutes and their solutions are neutral or
rothiazide is about 6 minutes.
slightly acidic.
System performance: When the procedure is run with 10 Loss on drying <2.41> 2 – 7z (1 g, 1059C, 2 hours).
ditions, hydrochlorothiazide and the internal standard are
and TYMC are 103 CFU/g and 102 CFU/g, respectively.
being not less than 5. Containers and storage Containers—Well-closed contain-
System repeatability: When the test is repeated 6 times ers.
the peak area of hydrochlorothiazide to that of the internal Pullulan Capsules
プルランカプセル
Pullulan Capsules are made of Pullulan as the base

Capsules material, and their shape is a pair of cylinders with one
end closed which can be overlapped on each other.
カプセル The label states the use or nonuse of the gelling
agent and its name.
Capsules are made of Gelatin, and their shape is a Method of preparation Dissolve Pullulan in water by
pair of cylinders with one end closed which can be warming, add, if necessary, emulsifiers, dispersing agents,
overlapped on each other. preservatives, coloring agents, gelling agents, and gelling
aid, etc. to make a viscous liquid, and form into a certain
Method of preparation Dissolve Gelatin in water by warm-
shape while warming.
ing, add Glycerin or D-Sorbitol, Macrogol 4000, emulsifier,
They may be coated with a lubricant as necessary.
dispersing agent, preservatives, coloring substances and so
forth, if necessary, to make a viscous liquid, and form into Solubility and acidity or alkalinity Place one pair of Pullu-
capsules while warm. lan Capsules without snapping in a 100-mL conical flask,
Capsules may be coated with a lubricant, if necessary. add 50 mL of water, and shake occasionally at 37 ± 29C.
When perform this test 5 times, either capsule dissolves
Solubility and acidity or alkalinity Place, without overlap-
within 10 minutes and these solutions are neutral or slightly
ping of the parts, 1 piece (1 pair) of Capsules in a 100–mL
acidic.
JP XVII Official Monographs / Carbamazepine 575
Loss on drying <2.41> 10 – 14z (1 g, 1059C, 6 hours). lution. Separately, dissolve 25 mg of 1,1?-[3,3?-dithiobis(2-
methyl-1-oxopropyl)]-L-diproline in methanol to make ex-
and TYMC are 103 CFU/g and 102 CFU/g, respectively.
Containers and storage Containers—Well-closed contain- tion and standard solution as directed under Liquid Chroma-
ers. tography <2.01> according to the following conditions, and
determine the peak areas, AT and AS, of 1,1?-[3,3?-dithio-
bis(2-methyl-1-oxopropyl)]-L-diproline in each solution: AT
Captopril is not larger than AS.
カプトプリル Detector: An ultraviolet absorption photometer (wave-
length: 220 nm).
259C.
C9H15NO3S: 217.29 Mobile phase: A mixture of water, methanol and phos-
(2S )-1-[(2S )-2-Methyl-3-sulfanylpropanoyl]pyrrolidine-2- phoric acid (1000:1000:1).
carboxylic acid Flow rate: Adjust so that the retention time of 1,1?-[3,3?-
[62571-86-2] dithiobis(2-methyl-1-oxopropyl)]-L-diproline is about 10
minutes.
Captopril contains not less than 98.0z of captopril System suitability—
(C9H15NO3S), calculated on the dried basis. System performance: Dissolve 25 mg each of Captopril
and 1,1?-[3,3?-dithiobis(2-methyl-1-oxopropyl)]-L-diproline
Description Captopril occurs as white, crystals or crystal-
in 200 mL of methanol. When the procedure is run with 20
line powder.
mL of this solution under the above operating conditions,
It is very soluble in methanol, freely soluble in ethanol
captopril and 1,1?-[3,3?-dithiobis(2-methyl-1-oxopropyl)]-
(99.5), and soluble in water.
L-diproline are eluted in this order with the resolution be-
Identification Determine the infrared absorption spectrum tween these peaks being not less than 3.
of Captopril as directed in the potassium bromide disk System repeatability: When the test is repeated 5 times
method under Infrared Spectrophotometry <2.25>, and com- with 20 mL of the standard solution under the above operat-
pare the spectrum with the Reference Spectrum: both spectra ing conditions, the relative standard deviation of the peak
exhibit similar intensities of absorption at the same wave areas of 1,1?-[3,3?-dithiobis(2-methyl-1-oxopropyl)]-L-dipro-
numbers. line is not more than 2.0z.
D : －125 – －1349(after drying, Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-
0.1 g, ethanol (99.5), 10 mL, 100 mm). um, 809C, 3 hours).
Melting point <2.60> 105 – 1109C Residue on ignition <2.44> Not more than 0.2z (1 g).
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Assay Weigh accurately about 0.3 g of Captopril, dissolve
Captopril according to Method 2, and perform the test. Pre- in 100 mL of water, add 20 mL of dilute sulfuric acid and 1 g
pare the control solution with 2.0 mL of Standard Lead So- of potassium iodide, and shake. Titrate <2.50> with 1/60
lution (not more than 20 ppm). mol/L potassium iodate VS (indicator: 2 mL of starch TS).
(2) Arsenic <1.11>—Prepare the test solution with 1.0 g Perform a blank determination in the same manner, and
of Captopril according to Method 1, and perform the test make any necessary correction.
Each mL of 1/60 mol/L potassium iodate VS
(3) Related substances—Dissolve 0.20 g of Captopril in
＝ 21.73 mg of C9H15NO3S
methanol to make exactly 10 mL, and use this solution as the
sample solution. Separately, dissolve 15 mg of 1,1?-[3,3?- Containers and storage Containers—Tight containers.
dithiobis(2-methyl-1-oxopropyl)]-L-diproline in methanol to
solution. Perform the test with these solutions as directed Carbamazepine
under Thin-layer Chromatography <2.03>. Spot 10 mL each
of the sample solution and standard solution on a plate of カルバマゼピン
silica gel for thin-layer chromatography. Develop with a
mixture of toluene and acetic acid (100) (13:7) to a distance
of about 15 cm, and air-dry the plate. Place the plate in a
chamber filled with iodine vapor, and allow to stand for 30
minutes: the number of the spots other than the spot cor-
responding to that from the standard solution and the princi-
pal spot from the sample solution is not more than two, and C15H12N2O: 236.27
they are not more intense than the spot from the standard 5H-Dibenz[b, f ]azepine-5-carboxamide
solution. [298-46-4]
(4) 1,1?-[3,3?-Dithiobis(2-methyl-1-oxopropyl)]-L-
diproline—Dissolive 0.10 g of Captopril in methanol to Carbamazepine, when dried, contains not less than
make exactly 20 mL, and use this solution as the sample so- 97.0z and not more than 103.0z of carbamazepine
576 Carbazochrome Sodium Sulfonate Hydrate / Official Monographs JP XVII
(C15H12N2O). (95) to make exactly 100 mL. Perform the test as directed
under Ultraviolet-visible Spectrophotometry <2.24>, and de-
Description Carbamazepine occurs as a white to slightly
termine the absorbance A of this solution at the wavelength
yellowish white powder. It is odorless and tasteless at first,
of maximum absorption at about 285 nm.
and leaves a slightly bitter aftertaste.
It is freely soluble in chloroform, sparingly soluble in Amount (mg) of carbamazepine (C15H12N2O)
ethanol (95) and in acetone, and very slightly soluble in ＝ A/490 × 50,000
water and in diethyl ether.
Identification (1) To 0.1 g of Carbamazepine add 2 mL
of nitric acid, and heat on a water bath for 3 minutes: an
orange-red color is produced. Carbazochrome Sodium Sulfonate
(2) To 0.1 g of Carbamazepine add 2 mL of sulfuric
acid, and heat on a water bath for 3 minutes: a yellow color Hydrate
is produced with a green fluorescence.
カルバゾクロムスルホン酸ナトリウム水和物
(3) Examine Carbamazepine under ultraviolet light: the
solution shows an intense blue fluorescence.
(4) Determine the absorption spectrum of the solution
obtained in the Assay as directed under Ultraviolet-visible
C10H11N4NaO5S.3H2O: 376.32
Monosodium (2RS )-1-methyl-6-oxo-5-semicarbazono-
Purity (1) Clarity and color of solution—Dissolve 1.0 g 2,3,5,6-tetrahydroindole-2-sulfonate trihydrate
of Carbamazepine in 10 mL of chloroform: the solution is [51460-26-5, anhydride]
clear and colorless to pale yellow.
(2) Acidity—To 2.0 g of Carbamazepine add exactly 40 Carbazochrome Sodium Sulfonate Hydrate contains
mL of water, stir well for 15 minutes, and filter through a not less than 98.0z and not more than 102.0z of
glass filter (G3). To 10 mL of this filtrate add 1 drop of carbazochrome sodium sulfonate (C10H11N4NaO5S:
phenolphthalein TS and 0.50 mL of 0.01 mol/L sodium hy- 322.27), calculated on the anhydrous basis.
droxide VS: a red color is produced.
Description Carbazochrome Sodium Sulfonate Hydrate
(3) Alkalinity—To 10 mL of the filtrate obtained in (2)
occurs as orange-yellow, crystals or crystalline powder.
add 1 drop of methyl red TS and 0.50 mL of 0.01 mol/L
It is sparingly soluble in water, very slightly soluble in
hydrochloric acid VS: a red color is produced.
methanol and in ethanol (95), and practically insoluble in
(4) Chloride <1.03>—Dissolve 0.25 g of Carbamazepine
diethyl ether.
in 30 mL of acetone, add 6 mL of dilute nitric acid and water
A solution of Carbazochrome Sodium Sulfonate Hydrate
(1 in 100) shows no optical rotation.
the test solution. Prepare the control solution as follows: to
0.20 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of
acetone, 6 mL of dilute nitric acid and water to make 50 mL Identification (1) Determine the absorption spectrum of a
(not more than 0.028z). solution of Carbazochrome Sodium Sulfonate Hydrate (1 in
(5) Heavy metals <1.07>—Proceed with 2.0 g of Car- 100,000) as directed under Ultraviolet-visible Spectropho-
bamazepine according to Method 2, and perform the test. tometry <2.24>, and compare the spectrum with the Refer-
Prepare the control solution with 2.0 mL of Standard Lead ence Spectrum: both spectra exhibit similar intensities of ab-
Solution (not more than 10 ppm). sorption at the same wavelengths.
(6) Related substances—Dissolve 0.25 g of Carbamaze- (2) Determine the infrared absorption spectrum of Car-
pine in 10 mL of chloroform, and use this solution as the bazochrome Sodium Sulfonate Hydrate as directed in the
sample solution. Separately, dissolve 5.0 mg of iminodiben- potassium bromide disk method under Infrared Spectropho-
zyl in chloroform to make exactly 100 mL, and use this solu- tometry <2.25>, and compare the spectrum with the Refer-
tion as the standard solution. Perform the test with these so- ence Spectrum: both spectra exhibit similar intensities of ab-
lutions as directed under Thin-layer Chromatography <2.03>. sorption at the same wave numbers.
Spot 10 mL each of the sample solution and standard solu- (3) A solution of Carbazochrome Sodium Sulfonate Hy-
tion on a plate of silica gel for thin-layer chromatography. drate (1 in 100) responds to the Qualitative Tests <1.09> (1)
Develop the plate with a mixture of toluene and methanol for sodium salt.
(19:1) to a distance of about 10 cm, and air-dry the plate.
pH <2.54> Dissolve 0.8 g of Carbazochrome Sodium Sul-
Spray evenly potassium dichromate-sulfuric acid TS on the
fonate Hydrate in 50 mL of water by warming, and cool: the
plate: the spots other than the principal spot obtained from
pH of this solution is between 5.0 and 6.0.
the sample solution is not more intense than the spot ob-
tained from the standard solution. Purity (1) Clarity and color of solution—Dissolve 1.0 g
of Carbazochrome Sodium Sulfonate Hydrate in 50 mL of
water by warming, and allow to cool: the solution is clear.
2 hours).
Perform the test with this solution as directed under Ultravi-
Residue on ignition <2.44> Not more than 0.1z (1 g). olet-visible Spectrophotometry <2.24>: the absorbance at 590
nm is not more than 0.070.
Assay Dissolve about 50 mg of Carbamazepine, previously
(2) Heavy metals <1.07>—Proceed with 1.0 g of Car-
dried and accurately weighed, in ethanol (95) to make ex-
bazochrome Sodium Sulfonate Hydrate according to
actly 250 mL. Pipet 5 mL of this solution and add ethanol
Method 2, and perform the test. Prepare the control solution
JP XVII Official Monographs / Carbidopa Hydrate 577
with 2.0 mL of Standard Lead Solution (not more than 20 Containers and storage Containers—Well-closed contain-
ppm). ers.
(3) Related substances—Dissolve 50 mg of Carbazo-
chrome Sodium Sulfonate Hydrate in 100 mL of water, and
use this solution as the sample solution. Pipet 2 mL of the Carbidopa Hydrate
sample solution, add water to make exactly 200 mL, and use
this solution as the standard solution. Perform the test with カルビドパ水和物
area of these solutions by the automatic integration method:
the total area of the peaks other than carbazochrome sul-
fonate from the sample solution is not larger than the peak
C10H14N2O4.H2O: 244.24
area of carbazochrome sulfonate from the standard solu-
(2S )-2-(3,4-Dihydroxybenzyl)-2-hydrazinopropanoic
tion.
acid monohydrate
[38821-49-7]
length: 360 nm).
Carbidopa Hydrate contains not less than 98.0z of
carbidopa hydrate (C10H14N2O4.H2O).
gel for liquid chromatography (7 mm in particle diameter). Description Carbidopa Hydrate occurs as a white to yel-
Column temperature: A constant temperature of about lowish white powder.
409 C. It is sparingly soluble in methanol, slightly soluble in
Mobile phase: Dissolve 1.2 g of ammonium dihydrogen water, very slightly soluble in ethanol (95), and practically
phosphate in 1000 mL of water, and filter through a mem- insoluble in diethyl ether.
brane filter (0.4 mm in pore size) if necessary. To 925 mL of Melting point: about 1979C (with decomposition).
this solution add 75 mL of ethanol (95), shake, and adjust
Identification (1) Dissolve 0.01 g of Carbidopa Hydrate
the pH to 3 with phosphoric acid.
in 250 mL of a solution of hydrochloric acid in methanol (9
Flow rate: Adjust so that the retention time of car-
in 1000). Determine the absorption spectrum of this solution
bazochrome sulfonate is about 7 minutes.
<2.24>, and compare the spectrum with the Reference
retention time of carbazochrome sulfonate, beginning after
Spectrum or the spectrum of a solution of Carbidopa RS
the solvent peak.
wavelengths.
(2) Determine the infrared absorption spectrum of Car-
mL. Confirm that the peak area of carbazochrome sulfonate
bidopa Hydrate as directed in the potassium bromide disk
13z of that of carbazochrome sulfonate obtained from 10
pare the spectrum with the Reference Spectrum or the spec-
trum of Carbidopa RS: both spectra exhibit similar intensi-
System performance: Dissolve 10 mg each of Carbazo-
chrome Sodium Sulfonate Hydrate and carbazochrome in
100 mL of water by warming. When the procedure is run Optical rotation <2.49> [a]20
D : －21.0 – －23.59(1 g, alumi-
with 10 mL of this solution under the above operating condi- num (III) chloride TS, 100 mL, 100 mm).
tions, carbazochrome sulfonate and carbazochrome are
Carbidopa Hydrate according to Method 2, and perform the
(2) Related substances—Dissolve 50 mg of Carbidopa
Hydrate in 70 mL of the mobile phase, by warming and
area of carbazochrome sulfonate is not more than 2.0z.
using ultrasonication, if necessary. After cooling, add the
Water <2.48> 13.0 – 16.0z (0.3 g, volumetric titration, mobile phase to make 100 mL, and use this solution as the
direct titration). sample solution. Pipet 1 mL of the sample solution, add the
Assay Weigh accurately about 0.25 g of Carbazochrome
Sodium Sulfonate Hydrate, dissolve in 50 mL of water,
apply to a chromatographic column, 10 mm in diameter,
previously prepared with 20 mL of strongly acidic ion
lowing conditions. Determine each peak area from both so-
exchange resin for column chromatography (type H), and
lutions by the automatic integration method: the total area
allow to flow at a rate of 4 mL per minute. Wash the column
of the peaks other than carbidopa from the sample solution
with 150 mL of water, combine the washing and the former
is not larger than the peak area of carbidopa from the stand-
effluent solution, and titrate <2.50> with 0.05 mol/L sodium
ard solution.
hydroxide VS (potentiometric titration). Perform a blank
Each mL of 0.05 mol/L sodium hydroxide VS flow rate: Proceed as directed in the operating conditions in
＝ 16.11 mg of C10H11N4NaO5S the Assay.
578 L-Carbocisteine / Official Monographs JP XVII
retention time of carbidopa. L-Carbocisteine
System performance, and system repeatability: Proceed as L-カルボシステイン
mL. Confirm that the peak area of carbidopa obtained from
20 mL of this solution is equivalent to 7 to 13z of that of C5H9NO4S: 179.19
carbidopa obtained from 20 mL of the standard solution. (2R)-2-Amino-3-carboxymethylsulfanylpropanoic acid
[638-23-3]
Loss on drying <2.41> 6.9 – 7.9z (1 g, in vacuum not
exceeding 0.67 kPa, 1009C, 6 hours).
L-Carbocisteine, when dried, contains not less than
Residue on ignition <2.44> Not more than 0.1z (1 g). 98.5z of L-carbocisteine (C5H9NO4S).
Assay Weigh accurately about 50 mg each of Carbidopa Description L-Carbocisteine occurs as a white crystalline
Hydrate and Carbidopa RS (separately determine the loss on powder. It is odorless, and has a slightly acid taste.
drying <2.41> under the same conditions as Carbidopa Hy- It is very slightly soluble in water, and practically insoluble
drate), and dissolve each in 70 mL of the mobile phase, by in ethanol (95).
warming and using ultrasonication if necessary. After cool- It dissolves in dilute hydrochloric acid or in sodium hy-
ing, add the mobile phase to make exactly 100 mL, and use droxide TS.
these solutions as the sample solution and the standard solu- Melting point: about 1869 C (with decomposition).
tion, respectively. Perform the test with exactly 20 mL each
Identification (1) To 0.2 g of L-Carbocisteine add 1 mL
of lead acetate TS and 3 mL of water, shake, add 0.2 g of so-
dium hydroxide, and heat over a flame for 1 minute: a dark
brown to black precipitate is formed.
of carbidopa in each solution.
(2) Determine the infrared absorption spectrum of L-
Amount (mg) of carbidopa hydrate (C10H14N2O4.H2O) Carbocisteine as directed in the potassium bromide disk
＝ MS × AT/AS × 1.080 method under Infrared Spectrophotometry <2.25>, and com-
MS: Amount (mg) of Carbidopa RS taken, calculated on
the dried basis
numbers.
Optical rotation <2.49> [a]20D : －33.5 – －36.59Weigh accu-
rately about 5 g of L-Carbocisteine, previously dried, dis-
length: 280 nm).
solve in 20 mL of water and a suitable amount of a solution
of sodium hydroxide (13 in 100), and adjust the pH with 1
mol/L hydrochloric acid TS or 0.1 mol/L hydrochloric acid
TS to 6.0, and add water to make exactly 50 mL. Determine
the optical rotation of this solution in a 100-mm cell.
259 C.
Mobile phase: To 950 mL of 0.05 mol/L sodium dihydro- Purity (1) Clarity and color of solution—Dissolve 1.0 g
gen phosphate TS add 50 mL of ethanol (95), and adjust the of L-Carbocisteine in 10 mL of sodium hydroxide TS: the so-
pH to 2.7 with phosphoric acid. lution is clear and colorless.
Flow rate: Adjust so that the retention time of carbidopa (2) Chloride <1.03>—Dissolve 0.20 g of L-Carbocisteine
is about 6 minutes. in 10 mL of water and 20 mL of nitric acid, and add water to
System suitability— make 50 mL. Perform the test using this solution as the test
System performance: Dissolve 50 mg each of Carbidopa solution. Prepare the control solution as follows. To 0.40
Hydrate and methyldopa in 100 mL of the mobile phase. mL of 0.01 mol/L hydrochloric acid VS add 20 mL of nitric
When the procedure is run with 20 mL of this solution under acid and water to make 50 mL (not more than 0.071z).
the above operating conditions, methyldopa and carbidopa (3) Ammonium <1.02>—Perform the test with 0.25 g of
are eluted in this order with the resolution between these L-Carbocisteine using the distillation under reduced pres-
peaks being not less than 0.9. sure. Prepare the control solution with 5.0 mL of Standard
System repeatability: When the test is repeated 6 times Ammonium Solution (not more than 0.02z).
with 20 mL of the standard solution under the above operat- (4) Heavy metals <1.07>—Proceed with 1.0 g of L-Carbo-
ing conditions, the relative standard deviation of the peak cisteine according to Method 2, and perform the test. Pre-
area of carbidopa is not more than 1.0z. pare the control solution with 2.0 mL of Standard Lead So-
of L-Carbocisteine according to Method 3, and perform the
test (not more than 2 ppm).
(6) Related substances—Dissolve 0.30 g of L-Carbo-
cisteine in 10 mL of 0.2 mol/L sodium hydroxide TS, and
use this solution as the sample solution. Pipet 2 mL of the
sample solution, and add 0.2 mol/L sodium hydroxide TS to
make exactly 100 mL. Pipet 1 mL of this solution, and add
0.2 mol/L sodium hydroxide TS to make exactly 10 mL, and
JP XVII Official Monographs / L-Carbocisteine Tablets 579
with these solutions as directed under Thin-layer Chroma- areas, AT and AS, of L-carbocisteine in each solution.
tography <2.03>. Spot 5 mL each of the sample solution and
standard solution, in 15 mm length along the starting line on
of L-carbocisteine (C5H9NO4S)
a plate of silica gel for thin-layer chromatography. Develop
the plate with a mixture of 1-butanol, water and acetic acid ＝ MS × AT/AS × V?/V × 1/C × 450
(100) (3:1:1) to a distance of about 10 cm, and dry the plate
MS: Amount (mg) of L-carbocisteine for assay taken
at 809C for 30 minutes. Spray evenly a solution of ninhydrin
C: Labeled amount (mg) of L-carbocisteine (C5H9NO4S) in
in acetone (1 in 50) on the plate, and heat at 809 C for 5
1 tablet
minutes: the spots other than the principal spot from the
sample solution are not more intense than the spot from the Operating conditions—
standard solution. Proceed as directed in the operating conditions in the
Assay.
C,
2 hours).
Residue on ignition <2.44> Not more than 0.1z (1 g). mL of the standard solution under the above operating con-
Assay Weigh accurately about 0.25 g of L-Carbocisteine,
factor of the peak of L-carbocisteine are not less than 1500
previously dried, dissolve in exactly 20 mL of 0.1 mol/L per-
chloric acid VS and 50 mL of acetic acid (100), and titrate
<2.50> the excess perchloric acid with 0.1 mol/L sodium ace-
tate VS (potentiometric titration). Perform a blank determi-
nation.
area of L-carbocisteine is not more than 1.0z.
Assay To 10 L-Carbocisteine Tablets add 220 mL of 0.5
＝ 17.92 mg of C5H9NO4S
mol/L hydrochloric acid TS, stir for 30 minutes, add 0.5
Containers and storage Containers—Tight containers. mol/L hydrochloric acid TS to make exactly 250 mL, and
stir additionally for 30 minutes. Filter this solution, discard
the first 20 mL of the filtrate, pipet 2 mL of the subsequent
L-Carbocisteine Tablets filtrate, add (V—50)/25 mL of 0.5 mol/L hydrochloric acid
TS, then add exactly V/25 mL of the internal standard solu-
L-カルボシステイン錠 tion, add water to make V mL so that each mL contains
about 0.4 mg of L-carbocisteine (C5H9NO4S), and use this
L-Carbocisteine Tablets contain not less than 95.0z
about 20 mg of L-carbocisteine for assay, previously dried at
and not more than 105.0z of the labeled amount of L-
1059C for 2 hours, add 2 mL of 0.5 mol/L hydrochloric acid
carbocisteine (C5H9NO4S: 179.19).
TS, and exactly 2 mL of the internal standard solution. Then
Method of Preparation Prepare as directed under Tablets, add water to dissolve to make 50 mL, and use this solution
with L-Carbocisteine. as the standard solution. Perform the test with 5 mL each of
Identification Powder L-Carbocisteine Tablets. To a por-
tion of the powder, equivalent to 0.18 g of L-Carbocisteine,
add 50 mL of water, stir for 10 minutes, and filter. To 5 mL
area of L-carbocisteine to that of the internal standard.
of the filtrate add 1 mL of ninhydrin TS, and heat in a water
bath for 3 minutes: a purple color develops. Amount (mg) of L-carbocisteine (C5H9NO4S) in 1 tablet
＝ MS × QT/QS × V/4
of the Mass variation test. MS: Amount of L-carbocisteine for assay taken
Dissolution <6.10> When the test is performed at 75 revolu- Internal standard solution—A solution of nicotinic acid (9 in
tions per minute according to the Paddle method, using 900 10,000).
mL of water as the dissolution medium, the dissolution rates Operating conditions—
in 15 minutes of 250-mg tablet and in 30 minutes of 500-mg Detector: An ultraviolet absorption photometer (wave-
tablet are not less than 80z and not less than 85z, respec- length: 240 nm).
tively. Column: A stainless steel column 4.6 mm in inside diame-
Start the test with 1 tablet of L-Carbocisteine Tablets, ter and 15 cm in length, packed with octadecylsilanized silica
withdraw not less than 20 mL of the medium at the specified gel for liquid chromatography (5 mm in particle diameter).
minute after starting the test, and filter through a membrane Column temperature: A constant temperature of about
filter with a pore size not exceeding 0.45 mm. Discard the 209C.
first 10 mL of the filtrate, pipet V mL of the subsequent Mobile phase: Diluted trifluoroacetic acid (1 in 1000).
filtrate, add the mobile phase to make exactly V? mL so Flow rate: Adjust so that the retention time of L-carbo-
that each mL contains about 0.14 mg of L-carbocisteine cisteine is about 2 minutes.
(C5H9NO4S), and use this solution as the sample solution. System suitability—
Separately, weigh accurately about 28 mg of L-carbocisteine System performance: When the procedure is run with 5 mL
for assay, previously dried at 1059 C for 2 hours, and dis- of the standard solution under the above operating condi-
solve in the mobile phase to make exactly 200 mL, and use tions, L-carbocisteine and the internal standard are eluted in
this solution as the standard solution. Perform the test with this order with the resolution between these peaks being not
exactly 20 mL each of the sample solution and standard solu- less than 4.
tion as directed under Liquid Chromatography <2.01> ac- System repeatability: When the test is repeated 6 times
cording to the following condition, and determine the peak with 5 mL of the standard solution under the above operating
580 Carbon Dioxide / Official Monographs JP XVII
conditions, the relative standard deviation of the ratio of the Volume (mL) of carbon dioxide (CO2)
peak area of L-carbocisteine to that of the internal standard ＝ volume (mL) of the sample － V (mL)
Containers and storage Containers—Cylinders.
Containers and storage Containers—Tight containers. Storage—Not exceeding 409C.
Carbon Dioxide Carboplatin

二酸化炭素カルボプラチン
CO2: 44.01
[124-38-9]
Carbon Dioxide contains not less than 99.5 volz of

carbon dioxide (CO2). C6H12N2O4Pt: 371.25
(SP-4-2)-Diammine[cyclobutan-1,1-dicarboxylato(2-)-O,O?]platinum
Description Carbon Dioxide is a colorless gas at room tem-
[41575-94-4]
perature and under atmospheric pressure. It is odorless.
A 1 mL volume of Carbon Dioxide dissolves in 1 mL of
Carboplatin contains not less than 98.5z and not
water, and the solution is slightly acid.
more than 101.0z of carboplatin (C6H12N2O4Pt), cal-
1000 mL of Carbon Dioxide at 09C and under a pressure
of 101.3 kPa weighs 1.978 g.
Description Carboplatin occurs as white, crystals or crys-
Identification (1) Pass 100 mL of Carbon Dioxide
talline powder.
through a carbon dioxide measuring detector tube: the detec-
It is sparingly soluble in water, and very slightly soluble in
tor tube is changed to a stipulated color tone by each detec-
ethanol (99.5).
tor tube, provided that the detector tube with a upper limit
of measurement of not less than 10z is used.
(2) Pass Carbon Dioxide into calcium hydroxide TS: a Identification (1) To 2 mL of a solution of Carboplatin
white precipitate is produced. Collect the precipitate, and (1 in 100) add 2 to 3 drops of diluted tin (II) chloride TS (1 in
add acetic acid (31): it dissolves with effervescence. 15), and allow to stand for 30 minutes: a yellowish brown
Purity (1) Acidity—Place 50 mL of freshly boiled and
cooled water in a Nessler tube, and pass 1000 mL of Carbon
Carboplatin as directed in the potassium bromide disk
Dioxide into it for 15 minutes through an introducing tube
about 1 mm in diameter extending to 2 mm from the bottom
of the Nessler tube, then add 0.10 mL of methyl orange TS:
trum of Carboplatin RS: both spectra exhibit similar intensi-
the solution is not more colored than the following control
solution.
Control solution: To 50 mL of freshly boiled and cooled pH <2.54> Dissolve 0.10 g of Carboplatin in 10 mL of
water in a Nessler tube add 0.10 mL of methyl orange TS water: the pH of this solution is 5.0 to 7.0.
and 1.0 mL of 0.01 mol/L hydrochloric acid VS.
Purity (1) 1,1-Cyclobutanedicarboxylic acid—Weigh ac-
(2) Hydrogen phosphide, hydrogen sulfide or reducing
curately about 40 mg of Carboplatin, dissolve in the mobile
organic substances—Place 25 mL of silver nitrate-ammonia
TS and 3 mL of ammonia TS in each of two Nessler tubes
A and B, and designate the solution in each tube as solution
of 1,1-cyclobutanedicarboxylic acid, and dissolve in the
A and solution B, respectively. Pass 1000 mL of Carbon Di-
mobile phase to make exactly 100 mL. Pipet 4 mL of this
oxide into solution A in the same manner as directed in (1):
the turbidity and color of this solution are the same as that
of solution B.
(3) Carbon monoxide—Pass a specified amount of Car-
bon Dioxide through a carbon monoxide measuring detector
according to the following conditions. Determine the peak
tube: the concentration of carbon monoxide is less than 15
areas, AT and AS, of 1,1-cyclobutanedicarboxylic acid in
ppm, provided that the passing amount (mL) of Carbon
each solution, and calculate the amount of 1,1-cyclo-
Dioxide is stipulated according to each detector tube.
butanedicarboxylic acid by the following formula: it is not
Assay Place 125 mL of a solution of potassium hydroxide more than 0.2z.
(1 in 2) in a gas pipet of suitable capacity. Measure exactly
Amount (z) of 1,1-cyclobutanedicarboxylic acid
about 100 mL of Carbon Dioxide in a 100-mL gas buret
＝ MS/MT × AT/AS × 8/5
filled with water. Force the entire volume of gas into the gas
pipet, and shake for 5 minutes. Draw some of the unab- MS: Amount (mg) of 1,1-cyclobutanedicarboxylic acid
sorbed gas into the gas buret, measure the volume, force the taken
residual back upon the surface of the liquid in the gas pipet, MT: Amount (mg) of Carboplatin taken
and repeat this procedure until a constant volume of the
residual reading is obtained. Determine the volume V (mL)
of the residual gas. Calculate the volume of the sample and
length: 220 nm).
V on the basis of the gas volume at 209 C and at 101.3 kPa.
JP XVII Official Monographs / Carboplatin 581
ter and 30 cm in length, packed with octadecylsilanized silica solution for system suitability test, and add water to make
gel for liquid chromatography (7 mm in particle diameter). exactly 20 mL. Confirm that the peak area of carboplatin
Column temperature: A constant temperature of about obtained with 10 mL of this solution is equivalent to 3.5 to
359 C. 6.5z of that obtained with 10 mL of the solution for system
Mobile phase: Dissolve 8.5 g of tetrabutylammonium suitability test.
hydrogensulfate in 80 mL of water, add 3.4 mL of phos- System repeatability: When the test is repeated 6 times
phoric acid, and adjust to pH 7.5 with a solution of sodium with 10 mL of the solution for system suitability test under
hydroxide (43 in 100). To 10 mL of this solution add 430 mL the above operating conditions, the relative standard devia-
of water and 60 mL of acetonitrile. tion of the peak area of carboplatin is not more than 2.0z.
Flow rate: Adjust so that the retention time of 1,1-cy-
clobutanedicarboxylic acid is about 5 minutes.
4 hours).
Test for required detectability: To exactly 2 mL of the Assay Weigh accurately about 25 mg each of Carboplatin
standard solution add the mobile phase to make exactly 10 and Carboplatin RS (separately determine the loss on drying
mL. Confirm that the peak area of 1,1-cyclobutanedicarbox- <2.41> under the same conditions as Carboplatin), dissolve
ylic acid obtained with 25 mL of this solution is equivalent to separately in water to make exactly 25 mL, and use these
14 to 26z of that obtained with 25 mL of the standard solu- solutions as the sample solution and the standard solution,
tion. respectively. Perform the test with exactly 10 mL each of the
System performance: Dissolve 25 mg each of 1,1-cyclo- sample solution and standard solution as directed under
butanedicarboxylic acid and cyclobutanecarboxylic acid in Liquid Chromatography <2.01> according to the following
100 mL of water. To 10 mL of this solution add the mobile conditions, and determine the peak areas, AT and AS, of
phase to make 25 mL. When the procedure is run with 25 mL carboplatin in each solution.
of this solution under the above operating conditions, cyclo-
Amount (mg) of carboplatin (C6H12N2O4Pt)
butanecarboxylic acid and 1,1-cyclobutanedicarboxylic acid
＝ MS × AT/AS
are eluded in this order with the resolution between these
peaks being not less than 3. MS: Amount (mg) of Carboplatin RS taken, calculated on
System repeatability: When the test is repeated 6 times the dried basis
area of 1,1-cyclobutanedicarboxylic acid is not more than
length: 220 nm).
2.0z.
(2) Related substances—Dissolve 25 mg of Carboplatin
ter and 25 cm in length, packed with phenylhexylsilanized
in 25 mL of water, and use this solution as the sample solu-
silica gel for liquid chromatography (5 mm in particle diame-
tion. Perform the test with 10 mL of the sample solution as
ter).
279C.
the automatic integration method. Calculate the amount of
Mobile phase A: Dissolve 8.5 g of tetrabutylammonium
the peaks by the area percentage method: the amount of the
hydrogensulfate in 80 mL of water, add 3.4 mL of phos-
peak, having the relative retention time of about 0.8 to car-
phoric acid, and adjust to pH 7.5 with a solution of sodium
boplatin, is not more than 0.25z, the amount of the peak
hydroxide (43 in 100). To 20 mL of this solution add water
other than carboplatin and the peak mentioned above is not
to make 1000 mL.
more than 0.1z, and the total amount of the peaks other
Mobile phase B: Dissolve 8.5 g of tetrabutylammonium
than carboplatin is not more than 0.5z.
hydrogensulfate in 80 mL of water, add 3.4 mL of phos-
Detector, column, column temperature, mobile phases A
hydroxide (43 in 100). To 20 mL of this solution add water
and B, and flow rate: Proceed as directed in the operating
to make 800 mL, and add 200 mL of acetonitrile.
conditions in the Assay.
Time after injection Mobile phase Mobile phase

Time after injection Mobile phase Mobile phase
of sample (min) A (volz) B (volz)
of sample (min) A (volz) B (volz)
0 – 15 100 0
0 – 15 100 0
15 – 35 100 → 0 0 → 100
15 – 35 100 → 0 0 → 100
35 – 50 0 100
System performance: To 9 mL of the standard solution
retention time of carboplatin, beginning after the solvent
add 1 mL of diluted hydrogen peroxide TS (1 in 60), and
peak.
allow to stand at room temperature for not less than 1 hour.
the above operating conditions, the resolution between the
peak of carboplatin and the peak having the relative reten-
Test for required detectability: To 1 mL of the sample
tion time about 0.93 to carboplatin is not less than 1.2.
solution add water to make 100 mL, and use this solution as
the solution for system suitability test. Pipet 1 mL of the
582 Carboplatin Injection / Official Monographs JP XVII
ing conditions, the relative standard deviation of the peak the following conditions, and determine each peak area by
area of carboplatin is not more than 1.0z. the automatic integration method. Calculate the amount of
these peaks by the area percentage method: the total amount
of the peaks other than carboplatin is not more than 2.0z.
Detector, column, column temperature, mobile phases A
and B, and flow rate: Proceed as directed in the operating
Carboplatin Injection conditions in the Assay under Carboplatin.
Flowing of mobile phase, and time span of measurement:
カルボプラチン注射液
Proceed as directed in the operating conditions in the Purity
(2) under Carboplatin.
Carboplatin Injection is an aqueous injection. System suitability—
It contains not less than 95.0z and not more System performance: Proceed as directed in the system
than 105.0z of the labeled amount of carboplatin suitability in the Assay under Carboplatin.
(C6H12N2O4Pt: 371.25). Test for required detectability, and system repeatability:
Proceed as directed in the system suitability in the Purity (2)
under Carboplatin.
tions, with Carboplatin.
Description Carboplatin Injection is a clear, colorless to
pale yellow liquid. Extractable volume <6.05> It meets the requirement.
Identification (1) To an amount of Carboplatin Injec- Foreign insoluble matter <6.06> Perform the test according
tion, equivalent to 20 mg of Carboplatin, add 2 to 3 drops of to Method 1: it meets the requirement.
diluted tin (II) chloride TS (1 in 15), and allow to stand for
30 minutes: a yellowish brown precipitate is formed.
ment.
(2) Evaporate to dryness a volume of Carboplatin Injec-
tion, equivalent to 10 mg of Carboplatin, in a water bath at Sterility <4.06> Perform the test according to the Mem-
not exceeding 309C under vacuum. Determine the infrared brane filtration method: it meets the requirement.
absorption spectrum of the residue as directed in the potas-
Assay To an exact volume of Carboplatin Injection,
equivalent to about 20 mg of carboplatin (C6H12N2O4Pt),
try <2.25>: it exhibits absorption at the wave numbers of
about 3270 cm－1, 2990 cm－1, 2960 cm－1, 1645 cm－1, 1610
cm－1, 1381 cm－1 and 1348 cm－1.
mg of Carboplatin RS (separately determine the loss on dry-
pH Being specified separately when the drug is granted ap- ing <2.41> under the same conditions as Carboplatine), dis-
proval based on the Law. solve in water to make exactly 25 mL, and use this solution
Purity (1) 1,1-Cyclobutanedicarboxylic acid—To an ex-
act volume of Carboplatin Injection, equivalent to 20 mg of
Carboplatin, add the mobile phase to make exactly 10 mL,
of carboplatin in each solution.
weigh accurately about 25 mg of 1,1-cyclobutanedicarboxyl-
ic acid, and dissolve in the mobile phase to make exactly 100 Amount (mg) of carboplatin (C6H12N2O4Pt)
mL. Pipet 4 mL of this solution, add the mobile phase to ＝ MS × AT/AS × 4/5
MS: Amount (mg) of Carboplatin RS taken, calculated on
the dried basis
Liquid Chromatography <2.01> according to the following Operating conditions—
conditions. Determine the peak areas, AT and AS, of 1,1- Detector: An ultraviolet absorption photometer (wave-
cyclobutanedicarboxylic acid in each solution, and calculate length: 230 nm).
the amount of 1,1-cyclobutanedicarboxylic acid by the fol- Column: A stainless steel column 4.0 mm in inside diame-
lowing formula: it is not more than 0.7z. ter and 25 cm in length, packed with octadecylsilanized silica
Amount (z) of 1,1-cyclobutanedicarboxylic acid
＝ MS × AT/AS × 1/25
359C.
MS: Amount (mg) of 1,1-cyclobutanedicarboxylic acid Mobile phase: Dissolve 8.5 g of tetrabutylammonium
taken hydrogensulfate in 80 mL of water, add 3.4 mL of phos-
hydroxide (43 in 100). To 10 mL of this solution add 880 mL
of water and 10 mL of acetonitrile.
Purity (1) under Carboplatin.
Flow rate: Adjust so that the retention time of carboplatin
is about 4 minutes.
Proceed as directed in the system suitability in the Purity
(1) under Carboplatin.
System performance: To a solution of 25 mg of carbopla-
(2) Related substances—To a volume of Carboplatin In-
tin in 20 mL of water add 2.5 mL of a solution of 65 mg of
jection, equivalent to 10 mg of Carboplatin, add water to
1,3-phenylenediamine hydrochloride in 50 mL of water, and
add water to make 25 mL. When the procedure is run with
Perform the test with 10 mL of the sample solution as di-
10 mL of this solution under the above operating conditions,
JP XVII Official Monographs / Carmellose Calcium 583
carboplatin and 1,3-phenylenediamine are eluted in this tant liquid. Wash the precipitate with three 10-mL portions
order with the resolution between these peaks being not less of water by centrifuge each time, combine the supernatant
than 2.0. liquid and the washings, and add water to make 100 mL.
System repeatability: When the test is repeated 6 times Filter this solution, discard the first 5 mL of the filtrate, take
with 10 mL of the standard solution under the above operat- 25 mL of the subsequent filtrate in a Nessler tube, add 1 mL
ing conditions, the relative standard deviation of the peak of dilute hydrochloric acid and water to make 50 mL, and
area of carboplatin is not more than 1.0z. use this solution as the test solution. Separately, to 1.5 mL
of 0.005 mol/L sulfuric acid VS add 1 mL of dilute hydro-
chloric acid and water to make 50 mL, and use this solution
as the control solution. To the test solution and the control
Shelf life 24 months after preparation. solution add 2 mL each of barium chloride TS, mix, and
allow to stand for 10 minutes. Compare the opalescence de-
veloped in both solutions against a black background by
Carmellose viewing downward or transversely. The opalescence in the
test solution is not more intense than that in the control solu-
Carboxymethylcellulose tion (not more than 0.72z).

(3) Heavy metals <1.07>—Proceed with 1.0 g of Car-
カルメロース mellose according to Method 2, and perform the test. Pre-
[9000-11-7] lution (not more than 20 ppm).
4 hours).
that are not harmonized are marked with symbols ( ). Residue on ignition <2.44> Not more than 1.5z (after dry-
ing, 1 g).
Carmellose is partly O-carboxymethylated cellulose. Containers and storage Containers—Tight containers.
Description Carmellose occurs as a white powder.
It is practically insoluble in ethanol (95).
It swells with water to form suspension. Carmellose Calcium
It becomes viscid in sodium hydroxide TS.
It is hygroscopic. Carboxymethylcellulose Calcium
カルメロースカルシウム
trum of Carmellose as directed in the potassium bromide
[9050-04-8]
wave numbers.
(2) The pH <2.54> of a suspension, obtained by shaking
1 g of Carmellose with 100 mL of water, is between 3.5 and
5.0.
Carmellose Calcium is the calcium salt of a polycar-
Purity (1) Chloride—Shake well 0.8 g of Carmellose with boxymethylether of cellulose.
50 mL of water, add 10 mL of sodium hydroxide TS to dis- Description Carmellose Calcium occurs as a white to yel-
solve, and add water to make 100 mL. Heat 20 mL of this
lowish white powder.
solution with 10 mL of dilute nitric acid in a water bath until
It is practically insoluble in ethanol (95) and in diethyl
a flocculent precipitate is produced, cool, centrifuge, and
ether.
take out the supernatant liquid. Wash the precipitate with
It swells with water to form a suspension.
three 10-mL portions of water by centrifuge each time, com-
The pH of a suspension, obtained by shaking 1.0 g of Car-
bine the supernatant liquid and the washings, and add water
mellose Calcium with 100 mL of water, is between 4.5 and
to make 100 mL. Take 25 mL of this solution in a Nessler
6.0.
tube, add 6 mL of dilute nitric acid and water to make 50
mL, and use this solution as the test solution. Separately, to
0.40 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of Identification (1) Shake thoroughly 0.1 g of Carmellose
dilute nitric acid and water to make 50 mL, and use this solu- Calcium with 10 mL of water, followed by 2 mL of sodium
tion as the control solution. To the test solution and the con- hydroxide TS, allow to stand for 10 minutes, and use this so-
trol solution add 1 mL each of silver nitrate TS, mix, and lution as the sample solution. To 1 mL of the sample solu-
allow to stand protected from light for 5 minutes. Compare tion add water to make 5 mL. To 1 drop of this solution add
the opalescence developed in both solutions against a black 0.5 mL of chromotropic acid TS, and heat in a water bath
background by viewing downward or transversely. The for 10 minutes: a red-purple color develops.
opalescence in the test solution is not more intense than that (2) Shake 5 mL of the sample solution obtained in (1)
in the control solution (not more than 0.36z). with 10 mL of acetone: a white, flocculent precipitate is
(2) Sulfate—Shake well 0.40 g of Carmellose with 25 mL produced.
of water, add 5 mL of sodium hydroxide TS to dissolve, (3) Shake 5 mL of the sample solution obtained in (1)
and add 20 mL of water. Heat this solution with 2.5 mL of with 1 mL of iron (III) chloride TS: a brown, flocculent
hydrochloric acid in a water bath until a flocculent precipi- precipitate is produced.
tate is produced, cool, centrifuge, and take out the superna- (4) Ignite 1 g of Carmellose Calcium to ash, dissolve the
584 Carmellose Sodium / Official Monographs JP XVII
residue in 10 mL of water and 6 mL of acetic acid (31), and It forms a viscid solution in water and in warm water.
filter, if necessary. Boil the filtrate, cool, and neutralize with It is hygroscopic.
ammonia TS: the solution responds to the Qualitative Tests
Identification (1) Dissolve 0.2 g of Carmellose Sodium in
<1.09> (1) and (3) for calcium salt.
20 mL of warm water with stirring, cool, and use this solu-
Purity (1) Alkalinity—Shake thoroughly 1.0 g of Carmel- tion as the sample solution. To 1 mL of the sample solution
lose Calcium with 50 mL of freshly boiled and cooled water, add water to make 5 mL. To 1 drop of this solution add 0.5
and add 2 drops of phenolphthalein TS: no red color de- mL of concentrated chromotropic acid TS, and heat in a
velops. water bath for 10 minutes: a red-purple color develops.
(2) Chloride <1.03>—Shake thoroughly 0.80 g of Car- (2) To 10 mL of the sample solution obtained in test (1)
mellose Calcium with 50 mL of water, add 10 mL of sodium add 1 mL of copper (II) sulfate TS: a blue flocculent precipi-
hydroxide TS to dissolved, add water to make 100 mL, and tate is produced.
use this solution as the sample solution. Heat 20 mL of the (3) To 3 g of Carmellose Sodium add 20 mL of methanol
sample solution with 10 mL of 2 mol/L nitric acid TS on a and 2 mL of dilute hydrochloric acid, boil gently on a water
water bath until a flocculent precipitate is produced. After bath for 5 minutes, and filter. Evaporate the filtrate to dry-
cooling, centrifuge, and take out the supernatant liquid. ness, and add 20 mL of water to the residue: the solution re-
Wash the precipitate with three 10-mL portions of water by sponds to the Qualitative Tests <1.09> for sodium salt.
centrifuging each time, combine the supernatant and the
pH <2.54> Add 1.0 g of Carmellose Sodium in small por-
washings, and add water to make 100 mL. Take 25 mL of
tions to 100 mL of warm water with stirring, dissolve, and
this solution, and add 1 mL of nitric acid and water to make
cool: the pH of this solution is between 6.0 and 8.0.
tion. Prepare the control solution with 0.40 mL of 0.01 Purity (1) Clarity and color of solution—Firmly attach a
mol/L hydrochloric acid VS (not more than 0.36z). glass plate of good quality 2 mm in thickness, to the bottom
(3) Sulfate <1.14>—Heat 10 mL of the sample solution of a glass column 250 mm in height, 25 mm in inner diameter
obtained in (2) with 1 mL of hydrochloric acid in a water and 2 mm in thickness. This is used as an outer tube. Simi-
bath until a flocculent precipitate is produced. Cool, centri- larly prepare an inner tube by attaching a glass plate of good
fuge, and take out the supernatant liquid. Wash the precipi- quality 2 mm in thickness to the bottom of a glass column
tate with three 10-mL portions of water by centrifuging each 300 mm in height, 15 mm in inner diameter and 2 mm in
time, combine the supernatant and the washings, and add thickness. Dissolve 1.0 g of Carmellose Sodium in 100 mL of
water to make 100 mL. Perform the test with 25 mL this so- water, pour this solution into the outer tube, and place on a
lution as the test solution. Prepare the control solution with piece of white paper on which 15 parallel black lines 1 mm in
0.42 mL of 0.005 mol/L sulfuric acid VS. To the test solu- width and 1 mm in interval are drawn. Moving the inner
tion and the control solution add 1 mL of 3 mol/L hydro- tube up and down and observing from the upper part, deter-
chloric acid TS and 3 mL of barium chloride TS, then add mine the height of the solution up to the lower edge of the
water to make 50 mL, and mix. Allow to stand for 10 inner tube when the distinction of the lines becomes impos-
minutes, and compare the turbidity of these solutions: the sible. Repeat the operation 3 times, and calculate the mean
turbidity obtained with the test solution is not more than value: it is larger than that calculated from the similar opera-
that obtained with the control solution (not more than tion, using the following control solution.
1.0z). Control solution: To 5.50 mL of 0.005 mol/L sulfuric acid
(4) Heavy metals <1.07>—Proceed with 1.0 g of Car- VS add 1 mL of dilute hydrochloric acid, 5 mL of ethanol
mellose Calcium according to Method 2, and perform the (95) and water to make 50 mL. Add 2 mL of barium chloride
test. Prepare the control solution with 2.0 mL of Standard TS, mix well, and allow to stand for 10 minutes. Shake well
Lead Solution (not more than 20 ppm). this solution before use.
(2) Chloride <1.03>—Dissolve 0.5 g of Carmellose So-
C,
dium in 50 mL of water, and use this solution as the sample
4 hours).
solution. Shake 10 mL of the sample solution with 10 mL of
Residue on ignition <2.44> 10 – 20z (after drying 1 g). dilute nitric acid, heat to produce a flocculent precipitate in a
Containers water bath, cool, and centrifuge. Separate the supernatant
and storage Containers—Tight containers.
liquid, wash the precipitate with three 10-mL portions of
water, centrifuging each time, combine the supernatant liq-
uid with the washings, and dilute with water to 200 mL. Per-
Carmellose Sodium form the test using 50 mL of this solution as the test solu-
Carboxymethylcellulose Sodium mol/L hydrochloric acid VS (not more than 0.640z).
(3) Sulfate <1.14>—Add 1 mL of hydrochloric acid to 10
カルメロースナトリウム
mL of the sample solution obtained in (2), shake well, heat
to produce a flocculent precipitate in a water bath, cool, and
[9004-32-4]
centrifuge. Separate the supernatant liquid, wash the precipi-
tate with three 10-mL portions of water, centrifuging each
Carmellose Sodium is the sodium salt of a polycar-
time, combine the washings with the supernatant liquid men-
boxymethylether of cellulose.
tioned above, and dilute to 50 mL with water. Take 10 mL
It, when dried, contains not less than 6.5z and not
of this solution, dilute with water to 50 mL, and perform the
more than 8.5z of sodium (Na: 22.99).
test using this solution as the test solution. Prepare the con-
Description Carmellose Sodium occurs as a white to yel- trol solution with 0.40 mL of 0.005 mol/L sulfuric acid VS
lowish white, powder or granules. It has no taste. (not more than 0.960z).
It is practically insoluble in methanol, in ethanol (95), in (4) Silicate—Weigh accurately about 1 g of Carmellose
acetic acid (100) and in diethyl ether. Sodium, ignite in a platinum dish, add 20 mL of dilute hy-
JP XVII Official Monographs / Croscarmellose Sodium 585
drochloric acid, cover with a watch glass, and boil gently for 100 mL of a solution of methylene blue (1 in 250,000), stir
30 minutes. Remove the watch glass, and evaporate on a well, and allow to stand: blue cotton-like precipitates ap-
water bath to dryness with the aid of a current of air. Con- pear.
tinue heating for further 1 hour, add 10 mL of hot water, stir (2) To 1 g of Croscarmellose Sodium add 50 mL of
well, and filter through a filter paper for quantitative analy- water, and stir well to make a suspension. To 1 mL of this
sis. Wash the residue with hot water, dry together with the suspension add 1 mL of water and 5 drops of fleshly pre-
filter paper after no turbidity is produced on the addition of pared solution of 1-naphtol in methanol (1 in 25), and gently
silver nitrate TS to the last washing, and then ignite to con- add 2 mL of sulfuric acid along a wall of the vessel: a red-
stant mass: the mass of the residue is not more than 0.5z. purple color appears at the zone of contact.
(5) Heavy metals <1.07>—Proceed with 1.0 g of Carmel- (3) The suspension obtained in (2) responds to the
lose Sodium according to Method 2, and perform the test. Qualitative Tests <1.09> (1) for sodium salt.
pH <2.54> To 1.0 g of Croscarmellose Sodium add 100 mL
of water, and stir for 5 minutes: the pH of the supernatant
(6) Arsenic <1.11>—To 1.0 g of Carmellose Sodium add
liquid is between 5.0 and 7.0.
20 mL of nitric acid, heat gently until it becomes fluid, cool,
add 5 mL of sulfuric acid, and heat until white fumes are Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
evolved. Add, if necessary, 5 mL of nitric acid after cooling, Croscarmellose Sodium according to Method 2, and perform
and heat again. Repeat this operation until the solution the test. Prepare the control solution with 2.0 mL of Stand-
becomes colorless or slightly yellow. After cooling, add 15 ard Lead Solution (not more than 10 ppm).
mL of a saturated solution of ammonium oxalate monohy- 　(2) Sodium chloride and sodium glycolate—The total
drate, and heat until white fumes are evolved again, cool, amount of sodium chloride and sodium glycolate is not more
and dilute with water to 25 mL. Take 5 mL of this solution than 0.5z, calculated on the dried basis.
as the test solution, and perform the test. The solution has (i) Sodium chloride: Weigh accurately about 5 g of
no more color than the following color standard. Croscarmellose Sodium, add 50 mL of water and 5 mL of
Color standard: Without using Carmellose Sodium, hydrogen peroxide (30), and heat on a water bath for 20
proceed in the same manner, then transfer 5 mL of this solu- minutes with occasional stirring. After cooling, add 100 mL
tion to a generator bottle, add exactly 2 mL of Standard of water and 10 mL of nitric acid, and titrate <2.50> with 0.1
Arsenic Solution, and proceed as directed for the test with mol/L silver nitrate VS (potentiometric titration). Perform a
the test solution (not more than 10 ppm). blank determination in the same manner and make any nec-
(7) Starch—Add 2 drops of iodine TS to 10 mL of the essary correction.
sample solution obtained in (2): no blue color develops.
C, ＝ 5.844 mg of NaCl
4 hours).
(ii) Sodium glycolate: Weigh accurately about 0.5 g of
Assay Weigh accurately about 0.5 g of Carmellose So- Croscarmellose Sodium, add 2 mL of acetic acid (100) and
dium, previously dried, add 80 mL of acetic acid (100), con- 5 mL of water, and stir for 15 minutes. Add gradually 50 mL
nect with a reflux condenser, and heat in an oil bath main- of acetone with stirring, then add 1 g of sodium chloride, stir
tained at 1309C for 2 hours. Cool, and titrate <2.50> with 0.1 for 3 minutes, and filter through a filter paper moistened
mol/L perchloric acid VS (potentiometric titration). Per- with acetone. Wash the residue thoroughly with 30 mL of
form a blank determination, and make any necessary correc- acetone, combine the filtrate and washings, add acetone to
tion. make exactly 100 mL, and use this solution as the sample
stock solution. Separately, dissolve 0.100 g of glycolic acid
in water to make 200 mL. Pipet 0.5 mL, 1 mL, 2 mL, 3 mL
＝ 2.299 mg of Na
and 4 mL of this solution, add water to make them exactly 5
Containers and storage Containers—Tight containers. mL, then add 5 mL of acetic acid (100) and acetone to make
exactly 100 mL, and designate them standard stock solution
(1), standard stock solution (2), standard stock solution (3),
Croscarmellose Sodium standard stock solution (4) and standard stock solution (5),
respectively. Pipet 2 mL each of the sample stock solution
クロスカルメロースナトリウム and the standard stock solutions (1), (2), (3), (4) and (5), and
heat them in a water bath for 20 minutes to evaporate
[74811-65-7] acetone. After cooling, add exactly 5 mL of 2,7-dihydrox-
ynaphthalene TS, mix, then add 15 mL of 2,7-dihydrox-
This monograph is harmonized with the European Phar- ynaphthalene TS, mix, cover the mouth of the vessels with
macopoeia and the U.S. Pharmacopeia. The parts of the text aluminum foil, and heat in a water bath for 20 minutes.
that are not harmonized are marked with symbol ( ). After cooling, add sulfuric acid to make exactly 25 mL, mix,
and designate them sample solution, standard solution (1),
Croscarmellose Sodium is the sodium salt of a cross- standard solution (2), standard solution (3), standard solu-
linked poly carboxymethylether of cellulose. tion (4) and standard solution (5), respectively. Separately,
Description to 10 mL of a mixture of water and acetic acid (100) (1:1)
Croscarmellose Sodium occurs as a white to
add acetone to make exactly 100 mL, and proceed with ex-
yellowish white powder.
actly 2 mL of this solution in the same manner for prepara-
It is practically insoluble in ethanol (99.5) and in diethyl
tion of the sample solution, and use the solution so obtained
ether.
as the blank solution. Determine the absorbances, AT, AS1,
It swells with water and becomes a suspension.
AS2, AS3, AS4 and AS5, of the sample solution and the stand-
ard solutions (1), (2), (3), (4) and (5), respectively, at 540 nm
Identification (1) To 1 g of Croscarmellose Sodium add as directed under Ultraviolet-visible Spectrophotometry
586 Carmofur / Official Monographs JP XVII
<2.24>, using the blank solution as the control. Determine
the amount (g) of glycolic acid, X, in 100 ml of the sample Carmofur
solution from the calibration curve obtained with the stand-
ard solutions, and calculate the amount of sodium glycolate カルモフール
by the following formula.
Amount (z) of sodium glycolate
＝ X/M × 100 × 1.289
M: Amount (g) of sample taken, calculated on the dried
basis
　(3) Water-soluble substance—Weigh accurately about
10 g of Croscarmellose Sodium, disperse in 800 mL of water C11H16FN3O3: 257.26
by stirring for 1 minute every 10 minutes during 30 minutes, 5-Fluoro-1-(hexylaminocarbonyl)uracil
and allow to stand for at most 1 hour to precipitate. Filter by [61422-45-5]
suction or centrifuge the clear upper portion, and weigh ac-
curately the mass of about 150 mL of the filtrate or superna- Carmofur, when dried, contains not less than 98.0z
tant liquid. Heat to concentrate this liquid avoiding to dry- of carmofur (C11H16FN3O3).
ness, then dry at 1059C for 4 hours, and weigh the mass of
Description Carmofur occurs as a white crystalline pow-
the residue accurately. Calculate the amount of the water-
der.
soluble substance by the following formula: not less than
It is very soluble in N, N-dimethylformamide, freely solu-
1.0z and not more than 10.0z.
ble in acetic acid (100), soluble in diethyl ether, sparingly
Amount (z) of water-soluble substance soluble in methanol and in ethanol (99.5), and practically in-
＝ 100 M3 (800 ＋ M1)/M1M2 soluble in water.
M1: Amount (g) of sample taken, calculated on the dried
basis Identification (1) Proceed with 5 mg of Carmofur as di-
M2: Amount (g) of the filtrate or supernatant liquid of rected under Oxygen Flask Combustion Method <1.06>,
about 150 mL using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide
M3: Amount (g) of the residue TS and 20 mL of water as the absorbing liquid, and prepare
the test solution: the test solution responds to the Qualitative
Precipitation test Put 75 mL of water in a 100-mL glass-
Tests <1.09> (2) for fluoride.
stoppered graduated cylinder, and add portion by portion
with 1.5 g of Croscarmellose Sodium divided into three por-
Carmofur in a mixture of methanol and phosphoric acid-
tions while shaking vigorously at each time. Then, add water
acetic acid-boric acid buffer solution (pH 2.0) (9:1) (1 in
to make 100 mL, shake until to get a homogenous disper-
sion, and allow to stand for 4 hours: the volume of the set-
tled layer is not less than 10.0 mL and not more than 30.0
mL.
Degree of substitution Weigh accurately about 1 g of (3) Determine the infrared absorption spectrum of Car-
Croscarmellose Sodium, put in a 500-mL glass-stoppered mofur, previously dried, as directed in the potassium bro-
conical flask, add 300 mL of sodium chloride TS, then add mide disk method under Infrared Spectrophotometry <2.25>,
25.0 mL of 0.1 mol/L sodium hydroxide VS, stopper, and and compare the spectrum with the Reference Spectrum:
allow to stand for 5 minutes with occasional shaking. Add 5 both spectra exhibit similar intensities of absorption at the
drops of m-cresol purple TS, then add exactly 15 mL of 0.1 same wave numbers.
mol/L hydrochloric acid VS using a buret, stopper the flask,
and shake. If the color of the solution is purple, add exactly
Carmofur according to Method 2, and perform the test. Pre-
1-mL portions of 0.1 mol/L hydrochloric acid VS using the
buret, with shaking each time, until the color of the solution
changes to yellow, then titrate <2.50> with 0.1 mol/L sodium
(2) Related substances—Dissolve 0.20 g of Carmofur in
hydroxide VS until the color changes from yellow to purple.
10 mL of a mixture of methanol and acetic acid (100) (99:1),
Perform a blank determination in the same manner. Calcu-
late the degrees of substitution of acid-carboxymethyl group
the sample solution, add a mixture of methanol and acetic
and sodium-carboxymethyl group, A and S: A ＋ S is not
acid (100) (99:1) to make exactly 500 mL, and use this solu-
less than 0.60 and not more than 0.85.
tion as the standard solution. Perform the test with these so-
A ＝ 1150M/(7102 － 412M － 80C ) lutions as directed under Thin-layer Chromatography <2.03>.
S ＝ (162 ＋ 58A)C/(7102—80C ) Spot 15 mL each of the sample solution and standard solu-
tion on a plate of silica gel with fluorescent indicator for
M: Amount (mmol) of sodium hydroxide needed to neu-
tralize 1 g of sample taken, calculated on the dried
of toluene and acetone (5:3) to a distance of about 12 cm,
basis
C: The value (z) obtained in Residue on ignition
C, from the sample solution are not more intense than the spot
6 hours). from the standard solution. After exposure of the plate to
bromine vapor for 30 second, spray evenly a solution of
Residue on ignition <2.44> 14.0 – 28.0z (after drying, 1 g).
fluorescein in ethanol (95) (1 in 2500): the spots other than
Containers and storage Containers—Tight containers. the principal spot from the sample solution are not more in-
JP XVII Official Monographs / Carumonam Sodium 587
tense than the spot from the standard solution. (2) Heavy metals <1.07>—Proceed with 2.0 g of Car-
teolol Hydrochloride according to Method 2, and perform
um, 509C, 3 hours).
Residue on ignition <2.44> Not more than 0.1z (1 g). (3) Arsenic <1.11>—Prepare the test solution with 1.0 g
of Carteolol Hydrochloride according to Method 3, and per-
Assay Weigh accurately about 0.5 g of Carmofur, previ-
ously dried, dissolve in 20 mL of N, N-dimethylformamide,
(4) Related substances—Dissolve 0.20 g of Carteolol Hy-
and titrate <2.50> with 0.1 mol/L tetramethylammonium hy-
drochloride in 10 mL of methanol, and use this solution as
droxide-methanol VS until the color of the solution changes
from yellow through blue-green to blue (indicator: 3 drops
add methanol to make exactly 100 mL. Pipet 1 mL of this
of thymol blue-dimethylformamide TS).
Each mL of 0.1 mol/L tetramethylammonium solution as the standard solution. Perform the test with these
hydroxide-methanol VS solutions as directed under Thin-layer Chromatography
＝ 25.73 mg of C11H16FN3O3 <2.03>. Spot 10 mL each of the sample solution and standard
of chloroform, methanol and ammonia solution (28)
(50:20:1) to a distance of about 12 cm, and air-dry the plate.
Carteolol Hydrochloride Examine under ultraviolet light (main wavelength: 254 nm):
the spots other than the principal spot from the sample solu-
カルテオロール塩酸塩
tion are not more intense than the spot from the standard so-
lution.
3 hours).
C16H24N2O3.HCl: 328.83 Assay Weigh accurately about 0.5 g of Carteolol Hydro-
5-[(2RS )-3-(1,1-Dimethylethyl)amino- chloride, previously dried, add 30 mL of acetic acid (100),
2-hydroxypropyloxy]-3,4-dihydroquinolin-2(1H )-one dissolve by heating on a water bath, and cool. After adding
monohydrochloride 70 mL of acetic anhydride, titrate <2.50> with 0.1 mol/L per-
[51781-21-6] chloric acid VS (potentiometric titration). Perform a blank
Carteolol Hydrochloride, when dried, contains
not less than 99.0z of carteolol hydrochloride ＝ 32.88 mg of C16H24N2O3.HCl
(C16H24N2O3.HCl).
Description Carteolol Hydrochloride occurs as white, crys
ers.
It is soluble in water, sparingly soluble in methanol, very
slightly soluble in ethanol (95) and in acetic acid (100), and
practically insoluble in diethyl ether. Carumonam Sodium
The pH of a solution of 1.0 g of Carteolol Hydrochloride
カルモナムナトリウム
in 100 mL of water is between 5.0 and 6.0.
The solution of Carteolol Hydrochloride (1 in 20) shows
Identification (1) Dissolve 0.1 g of Carteolol Hydrochlo-
ride in 5 mL of water, and add 5 drops of Reinecke salt TS: a
light red precipitate is formed.
(2) Determine the absorption spectrum of a solution of C12H12N6Na2O10S2: 510.37
Carteolol Hydrochloride (1 in 100,000) as directed under Ul- Disodium (Z )-{(2-aminothiazol-4-yl)[(2S,3S )-2-
traviolet-visible Spectrophotometry <2.24>, and compare the carbamoyloxymethyl-4-oxo-1-sulfonatoazetidin-3-
spectrum with the Reference Spectrum: both spectra exhibit ylcarbamoyl]methyleneaminooxy}acetate
similar intensities of absorption at the same wavelengths. [86832-68-0]
teolol Hydrochloride as directed in the potassium chloride Carumonam Sodium contains not less than 850 mg
disk method under Infrared Spectrophotometry <2.25>, and (potency) and not more than 920 mg (potency) per mg,
compare the spectrum with the Reference Spectrum: both calculated on the anhydrous basis. The potency of
spectra exhibit similar intensities of absorption at the same Carumonam Sodium is expressed as mass (potency) of
wave numbers. carumonam (C12H14N6O10S2: 466.40).
(4) A solution of Carteolol Hydrochloride (1 in 50) re-
Description Carumonam Sodium occurs as a white to yel-
lowish white, crystals or crystalline powder.
Purity (1) Clarity and color of solution—Dissolve 1.0 g It is freely soluble in water, soluble in formamide, very
of Carteolol Hydrochloride in 30 mL of water: the solution slightly soluble in methanol, and practically insoluble in
is clear and colorless. ethanol (99.5) and in acetic acid (100).
588 Carumonam Sodium / Official Monographs JP XVII
Identification (1) Determine the absorption spectrum of a AS: Peak area of carumonam from the standard solution
solution of Carumonam Sodium (3 in 100,000) as directed AT: Each peak area other than carumonam from the sam-
under Ultraviolet-visible Spectrophotometry <2.24>, and ple solution
spectrum of a solution of Carumonam Sodium RS prepared
in the same manner as the sample solution: both spectra
exhibit similar intensities of absorption at the same wave-
the Assay.
lengths.
retention time of carumonam.
Carumonam Sodium as directed in the potassium bromide
the standard solution, and add the mobile phase to make ex-
spectrum of Carumonam Sodium RS: both spectra exhibit
actly 50 mL. Confirm that the peak area of carumonam ob-
(3) Determine the 1H spectrum of a solution of
of that obtained from 10 mL of the standard solution.
Carumonam Sodium in heavy water for nuclear magnetic
System performance: Dissolve 40 mg of Carumonam So-
resonance spectroscopy (1 in 10) as directed under Nuclear
dium in 20 mL of the mobile phase. To 5 mL of this solution
Magnetic Resonance Spectroscopy <2.21>, using sodium 3-
add 5 mL of a solution of resorcinol in the mobile phase (9
trimethylsilylpropionate-d4 for nuclear magnetic resonance
in 1000) and the mobile phase to make 25 mL. When the
spectroscopy as an internal reference compound: it exhibits a
double signal A at around d 5.5 ppm, and a single signal B at
operating conditions, resorcinol and carumonam are eluted
around d 7.0 ppm. The ratio of the integrated intensity of
these signals, A:B, is about 1:1.
not less than 2.5.
(4) Carumonam Sodium responds to the Qualitative
Tests <1.09> (1) for sodium salt.
D : ＋18.5 – ＋21.09(0.1 g calcu- ing conditions, the relative standard deviation of the peak
lated on the anhydrous basis, water, 10 mL, 100 mm). area of carumonam is not more than 2.0z.
(5) Related substance 2—Weigh accurately about 0.1 g
of Carumonam Sodium, and dissolve in the mobile phase to
1.0 g of Carumonam Sodium in 10 mL of water is between
5.0 and 6.5.
Purity (1) Clarity and color of solution—Dissolve 0.5 g the sample solution. Separately, weigh accurately about 0.1 g
of Carumonam Sodium in 5 mL of water: the solution is of Carumonam Sodium RS, and dissolve in the mobile phase
clear and colorless to pale yellow. to make exactly 50 mL. Pipet 5 mL of this solution, and add
(2) Heavy metals <1.07>—Proceed with 2.0 g of the mobile phase to make exactly 25 mL. Pipet 1 mL of this
Carumonam Sodium according to Method 2, and perform solution, add the mobile phase to make exactly 100 mL, and
the test. Prepare the control solution with 3.0 mL of Stand- use this solution as the standard solution. Perform the test
ard Lead Solution (not more than 15 ppm). with exactly 10 mL each of the sample solution and standard
(3) Arsenic <1.11>—Prepare the test solution with 2.0 g solution as directed under Liquid Chromatography <2.01>
of Carumonam Sodium according to Method 4, and perform according to the following conditions, and determine each
the test (not more than 1 ppm). peak area by the automatic integration method. Calculate
(4) Related substance 1—Weigh accurately about 0.1 g the amount of the related substances by the following equa-
of Carumonam Sodium, and dissolve in the mobile phase to tion: the amount of each related substance is not more than
make exactly 50 mL. Pipet 5 mL of this solution, add the 1.0z.
Amount (z) of related substance
the sample solution. Separately, weigh accurately about 0.1 g
＝ MS/MT × AT/AS
of Carumonam Sodium RS, and dissolve in the mobile phase
to make exactly 50 mL. Pipet 5 mL of this solution, and add MS: Amount (g) of Carumonam Sodium RS taken
the mobile phase to make exactly 25 mL. Pipet 1 mL of this MT: Amount (g) of Carumonam Sodium taken
solution, add the mobile phase to make exactly 100 mL, and AS: Peak area of carumonam from the standard solution
use this solution as the standard solution. Perform the test AT: Each area of the peaks appeared after the peak of
with exactly 10 mL each of the sample solution and standard carumonam from the sample solution
peak area by the automatic integration method. Calculate
directed in the operating conditions in the Assay.
the amount of the related substances by the following equa-
Mobile phase: A mixture of a solution of ammonium sul-
tion: the amount of the related substance having the relative
fate (1 in 10,000), methanol and acetic acid (100) (74:25:1).
retention time of about 0.7 to carumonam is not more than
Flow rate: Dissolve 0.01 g of phthalic acid in the mobile
4.0z, and each amount of the related substances other than
phase to make 100 mL. Adjust so that the retention time of
the related substance having the relative retention time of
phthalic acid is about 6.5 minutes when the procedure is run
about 0.7 to carumonam is not more than 1.0z.
with 10 mL of this solution.
Amount (z) of related substance Time span of measurement: About 10 times as long as the
＝ M S / M T × AT / AS retention time of carumonam.
MS: Amount (g) of Carumonam Sodium RS taken
MT: Amount (g) of Carumonam Sodium taken
JP XVII Official Monographs / Carvedilol 589
actly 50 mL. Confirm that the peak area of carumonam ob- Containers and storage Containers—Hermetic containers.
tained from 10 mL of this solution is equivalent to 7 to 13z Storage—Light-resistant.
System performance: Dissolve 40 mg of Carumonam So-
dium in 20 mL of the mobile phase. To 5 mL of this solution Carvedilol
add 5 mL of a solution of resorcinol in the mobile phase
(9 in 1000) and the mobile phase to make 25 mL. When the カルベジロール
operating conditions, resorcinol and carumonam are eluted
not less than 7.
area of carumonam is not more than 2.0z. C24H26N2O4: 406.47
(6) Total amount of related substances—The total of the (2RS )-1-(9H-Carbazol-4-yloxy)-
amounts of the related substances obtained in the Related 3-{[2-(2-methoxyphenoxy)ethyl]amino}propan-2-ol
substance 1 and the Related substance 2 is not more than [72956-09-3]
6.0z.
Carvedilol, when dried, contains not less than
99.0z and not more than 101.0z of carvedilol
tion, direct titration; Use a mixture of formamide for water
(C24H26N2O4).
determination and methanol for water determination (3:1)
instead of methanol for water determination). Description Carvedilol occurs as white to pale yellowish
Assay Weigh accurately an amount of Carumonam So-
It is freely soluble in acetic acid (100), sparingly soluble in
dium and Carumonam Sodium RS, equivalent to about 40
methanol, slightly soluble in ethanol (99.5), and practically
mg (potency), and dissolve each in the mobile phase to make
insoluble in water.
exactly 20 mL. Measure exactly 5 mL each of these solu-
A solution of Carvedilol in methanol (1 in 100) shows no
tions, add exactly 5 mL of the internal standard solution and
optical rotation.
the sample solution and standard solution. Perform the test Identification (1) Determine the absorption spectrum of a
with 10 mL each of the sample solution and standard solution solution of Carvedilol in methanol (1 in 200,000) as directed
as directed under Liquid Chromatography <2.01> according under Ultraviolet-visible Spectrophotometry <2.24>, and
to the following conditions, and calculate the ratios, QT and compare the spectrum with the Reference Spectrum: both
QS, of the peak area of carumonam to that of the internal spectra exhibit similar intensities of absorption at the same
standard. wavelengths.
Amount [ mg (potency)] of carumonam (C12H14N6O10S2)
vedilol as directed in the potassium bromide disk method
＝ MS × QT/QS × 1000
MS: Amount [mg (potency)] of Carumonam Sodium RS spectrum with the Reference Spectrum: both spectra exhibit
taken similar intensities of absorption at the same wave numbers.
Internal standard solution—A solution of resorcinol in the Melting point <2.60> 114 – 1199C
Purity (1) Heavy metals <1.07>—Wrap 2.0 g of Car-
vedilol with a filter paper for quantitative analysis, then
proceed according to Method 4, and perform the test. Pre-
length: 254 nm).
pare the control solution as follows: Put a filter paper for
quantitative analysis in a crucible, add 10 mL of a solution
of magnesium nitrate hexahydrate in ethanol (95) (1 in 10),
then proceed as directed for the test solution, and add 2.0
mL of Standard Lead Solution and water to make 50 mL
259 C.
Mobile phase: A mixture of a solution of ammonium sul-
(2) Related substances—Dissolve 65 mg of Carvedilol in
fate (1 in 10,000), methanol and acetic acid (100) (97:2:1).
100 mL of the mobile phase. To 1 mL of this solution add
Flow rate: Adjust so that the retention time of carumon-
the mobile phase to make 10 mL, and use this solution as the
am is about 10 minutes.
ditions, the internal standard and carumonam are eluted in
less than 2.5.
matic integration method: the area of the peak other than
carvedilol obtained from the sample solution is not larger
than 3/20 times the peak area of carvedilol obtained from
of the peak area of carumonam to that of the internal stand-
than carvedilol from the sample solution is not larger than
1/2 times the peak area of carvedilol from the standard solu-
590 Carvedilol Tablets / Official Monographs JP XVII
tion. tween 330 nm and 334 nm.
Purity Related substances—In this procedure the sample
solution should be stored not exceeding 59C and used within
length: 240 nm).
24 hours after preparation. Powder Carvedilol Tablets. Dis-
solve a portion of the powder, equivalent to 12.5 mg of Car-
vedilol, add an adequate amount of the mobile phase and
disperse the particles with the aid of ultrasonic waves, if nec-
essary, add the mobile phase to make 100 mL, and shake for
559 C.
30 minutes. Filter through a membrane filter with a pore size
not exceeding 0.22 mm, discard the first 5 mL of the filtrate,
phosphate in 900 mL of water, adjust to pH 2.0 with phos-
and use the subsequent filtrate as the sample solution. Pipet
phoric acid, and add water to make 1000 mL. To 650 mL of
1 mL of the sample solution, add the mobile phase to make
this solution add 350 mL of acetonitrile.
Flow rate: Adjust so that the retention time of carvedilol is
about 4 minutes.
retention time of carvedilol, beginning after the solvent
tions, and determine each peak area by the automatic in-
peak.
tegration method: the area of the two peaks, having the rela-
tive retention time between 1.7 and 1.9 and between 2.0 and
3.1 to carvedilol, obtained from the sample solution of
1.25-mg or 2.5-mg tablet is not larger than 3/10 times and
firm that the peak area of carvedilol obtained with 20 mL of
1.6 times the peak area of carvedilol obtained from the
this solution is equivalent to 7 to 13z of that obtained with
standard solution, respectively, the area of the peak other
20 mL of the standard solution.
than carvedilol and the peaks mentioned above from the
carvedilol from the standard solution, and the total area of
the peaks other than carvedilol from the sample solution is
factor of the peak of carvedilol are not less than 6000 and
not larger than 2.2 times the peak area of carvedilol from the
standard solution. The area of the two peaks, having the
relative retention time between 1.7 and 1.9 and between 2.0
and 3.1 to carvedilol, obtained from the sample solution of
10-mg or 20-mg tablet is not larger than 1/10 times and 2/5
area of carvedilol is not more than 2.0z.
times the peak area of carvedilol from the standard solution,
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, respectively, the area of the peak other than carvedilol and
2 hours). the peak mentioned above from the sample solution is not
larger than 1/10 times the peak area of carvedilol from the
Assay Weigh accurately about 0.5 g of Carvedilol, previ- carvedilol from the sample solution is not larger than 3/5
ously dried, dissolve in 60 mL of acetic acid (100), and titrate times the peak area of carvedilol from the standard solution.
<2.50> with 0.1 mol/L perchloric acid VS (potentiometric For the area of the peak, having the relative retention time
titration). Perform the blank determination in the same between 1.7 and 1.9 to carvedilol, multiply the relative
manner, and make any necessary correction. response factor 1.25.
＝ 40.65 mg of C24H26N2O4
Containers and storage Containers—Tight containers. the Assay.
retention time of carvedilol, beginning after the solvent
Carvedilol Tablets peak.
カルベジロール錠 Test for required detectability: To exactly 5 mL of the
mL. Confirm that the peak area of carvedilol obtained with
Carvedilol Tablets contain not less than 95.0z and
50 mL of this solution is equivalent to 3.5 to 6.5z of that
not more than 105.0z of the labeled amount of car-
obtained with 50 mL of the standard solution.
vedilol (C24H26N2O4: 406.47).
Method of preparation Prepare as directed under Tablets, mL of the standard solution under the above operating con-
with Carvedilol. ditions, the number of theoretical plates and the symmetry
factor of the peak of carvedilol are not less than 3000 and
Identification Powder Carvedilol Tablets. To a portion of
the powder, equivalent to 20 mg of Carvedilol, add 10 mL of
methanol, shake well, and filter. To 0.5 mL of the filtrate
area of carvedilol is not more than 1.0z.
visible Spectophotometry <2.24>: it exhibits maxima between
222 nm and 226 nm, between 241 nm and 245 nm, between Uniformity of dosage units <6.02> Perform the test accord-
284 nm and 288 nm, between 317 nm and 321 nm and being to the following method: it meets the requirement of the
JP XVII Official Monographs / Carvedilol Tablets 591
Content uniformity test. mL contains about 1.4 mg of carvedilol (C24H26N2O4), and

To 1 tablet of Carvedilol Tablets add 70 mL of a mixture use this solution as the sample solution. Separately, weigh
of 0.1 mol/L hydrochloric acid TS and methanol (1:1), accurately about 28 mg of carvedilol for assay, previously
shake until the tablet is completely disintegrated, then add a dried at 1059 C for 2 hours, and dissolve in methanol to
mixture of 0.1 mol/L hydrochloric acid TS and methanol make exactly 200 mL. Pipet 2 mL of this solution, add the
(1:1) to make exactly 100 mL, and filter through a mem- dissolution medium to make exactly 200 mL, and use this so-
brane filter with a pore size not exceeding 0.45 mm. Discard lution as the standard solution. Determine the absorbances,
the first 10 mL of the filtrate, pipet V mL of the subsequent AT and AS, at 240 nm of the sample solution and standard
filtrate, add a mixture of 0.1 mol/L hydrochloric acid TS solution as directed under Ultraviolet-visible Spectropho-
and methanol (1:1) to make exactly V? mL so that each mL tometry <2.24>, using the dissolution medium as the blank.
contains about 5 mg of carvedilol (C24H26N2O4), and use this
of carvedilol (C24H26N2O4)
about 25 mg of carvedilol for assay, previously dried at
＝ MS × AT/AS × V?/V × 1/C × 9/2
1059C for 2 hours, and dissolve in a mixture of 0.1 mol/L
hydrochloric acid TS and methanol (1:1) to make exactly 100 MS: Amount (mg) of carvedilol for assay taken
mL. Pipet 2 mL of this solution, add a mixture of 0.1 mol/L C: Labeled amount (mg) of carvedilol (C24H26N2O4) in 1
hydrochloric acid TS and methanol (1:1) to make exactly 100 tablet
Assay Weigh accurately the mass of not less than 20 Car-
vedilol Tablets, and powder. Weigh accurately a portion
of the powder, equivalent to about 25 mg of carvedilol
(C24H26N2O4), add exactly 5 mL of the internal standard so-
Amount (mg) of carvedilol (C24H26N2O4) lution, add a mixture of 0.1 mol/L hydrochloric acid TS and
＝ MS × AT/AS × V?/V × 1/50 methanol (1:1) to make 250 mL, and shake for 30 minutes.
To 2 mL of this solution, add the mobile phase to make 20
MS: Amount (mg) of carvedilol for assay taken
mL, and filter through a membrane filter with a pore size
Dissolution <6.10> (1) 10-mg tablet and 20-mg tablet　 not exceeding 0.45 mm. Discard the first 10 mL of the fil-
When the test is performed at 75 revolutions per minute trate, and use the subsequent filtrate as the sample solution.
according to the Paddle method, using 900 mL of 0.05 Separately, weigh accurately about 25 mg of carvedilol for
mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as assay, previously dried at 1059C for 2 hours, add exactly 5
the dissolution medium, the dissolution rate in 30 minutes of mL of the internal standard solution, and add a mixture of
Carvedilol Tablets is not less than 80z. 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make
Start the test with 1 tablet of Carvedilol Tablets, withdraw 250 mL. To 2 mL of this solution add the mobile phase to
not less than 20 mL of the medium at the specified minute make 20 mL, and use this solution as the standard solution.
after starting the test, and filter through a membrane filter Perform the test with 10 mL each of the sample solution and
with a pore size not exceeding 0.45 mm. Discard the first 10 standard solution as directed under Liquid Chromatography
mL of the filtrate, pipet V mL of the subsequent filtrate, add <2.01> under the following conditions, and calculate the
the dissolution medium to make exactly V? mL so that each ratios, QT and QS, of the peak area of carvedilol to that of
mL contains about 11 mg of carvedilol (C24H26N2O4), and use the internal standard.
Amount (mg) of carvedilol (C24H26N2O4)
rately about 28 mg of carvedilol for assay, previously dried
＝ M S × Q T / QS
at 1059 C for 2 hours, and dissolve in methanol to make ex-
actly 50 mL. Pipet 2 mL of this solution, add the dissolution MS: Amount (mg) of carvedilol for assay taken
medium to make exactly 100 mL, and use this solution as the
Internal standard solution—A solution of isoamyl parahy-
droxybenzoate in the mobile phase (1 in 70).
285 nm of the sample solution and standard solution as di-
using the dissolution medium as the blank.
length: 240 nm).
Dissolution rate (z) with respect to the labeled amount Column: A stainless steel column 4.6 mm in inside diame-
of carvedilol (C24H26N2O4) ter and 15 cm in length, packed with octadecylsilanized silica
＝ MS × AT/AS × V?/V × 1/C × 36 gel for liquid chromatography (5 mm in particle diameter).
MS: Amount (mg) of carvedilol for assay taken
409C.
C: Labeled amount (mg) of carvedilol (C24H26N2O4) in 1
tablet
(2) 1.25-mg tablet and 2.5-mg tablet When the test is with a solution prepared by dissolving 0.7 g of dipotassium
performed at 50 revolutions per minute according to the hydrogen phosphate in water to make 200 mL. To 450 mL of
Paddle method, using 900 mL of 0.05 mol/L acetic acid- this solution add 550 mL of methanol.
sodium acetate buffer solution (pH 4.0) as the dissolution Flow rate: Adjust so that the retention time of carvedilol is
medium, the dissolution rate in 20 minutes is not less than about 5 minutes.
75z. System suitability—
Start the test with 1 tablet of Carvedilol Tablets, withdraw System performance: When the procedure is run with 10
not less than 20 mL of the medium at the specified minute mL of the standard solution under the above operating con-
after starting the test, and filter through a membrane filter ditions, carvedilol and the internal standard are eluted in this
with a pore size not exceeding 0.45 mm. Discard the first 10 order with the resolution between these peaks being not less
mL of the filtrate, pipet V mL of the subsequent filtrate, add than 20.
the dissolution medium to make exactly V? mL so that each System repeatability: When the test is repeated 6 times
592 Cefaclor / Official Monographs JP XVII
with 10 mL of the standard solution under the above operat- (3) Related substances—Dissolve 50 mg of Cefaclor in 10
ing conditions, the relative standard deviation of the ratio of mL of sodium dihydrogen phosphate TS (pH 2.5), and use
the peak area of carvedilol to that of the internal standard is this solution as the sample solution. Pipet 1 mL of the sam-
not more than 1.0z. ple solution, add sodium dihydrogen phosphate TS (pH 2.5)
ard solution. Perform the test with exactly 20 mL each of the
Cefaclor ditions, and determine each peak area by the automatic in-
tegration method: the area of the peaks other than cefaclor
セファクロル
from the sample solution are not larger than 1/2 times the
peak area of cefaclor from the standard solution, and the
total area of the peaks other than cefaclor from the sample
solution is not larger than 2 times of the peak area of
cefaclor from the standard solution. If necessary, proceed
with 20 mL of sodium dihydrogen phosphate TS (pH 2.5) in
the same manner as above to compensate the base line.
C15H14ClN3O4S: 367.81 Operating conditions—
(6R,7R)-7-[(2R)-2-Amino-2-phenylacetylamino]-3- Detector: An ultraviolet absorption photometer (wave-
chloro-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- length: 220 nm).
carboxylic acid Column: A stainless steel column 4.6 mm in inside diame-
[53994-73-3] ter and 25 cm in length, packed with octadecylsilanized silica
Cefaclor contains not less than 950 mg (potency) Column temperature: A constant temperature of about
and not more than 1020 mg (potency) per mg, calcu- 259C.
lated on the anhydrous basis. The potency of Cefaclor Mobile phase A: Dissolve 7.8 g of sodium dihydrogen
is expressed as mass (potency) of cefaclor phosphate dihydrate in 1000 mL of water, and adjust the pH
(C15H14ClN3O4S). to 4.0 with phosphoric acid.
Mobile phase B: To 550 mL of the mobile phase A add
Description Cefaclor occurs as a white to yellowish white
450 mL of acetonitrile for liquid chromatography.
crystalline powder.
It is slightly soluble in water and in methanol, and practi-
cally insoluble in N, N-dimethylformamide and in ethanol
(99.5).
Time after injection Mobile phase A Mobile phase B
Identification (1) Determine the absorption spectrum of a of sample (min) (volz) (volz)
solution of Cefaclor (1 in 50,000) as directed under Ultravio-
let-visible Spectrophotometry <2.24>, and compare the spec- 0 – 30 95 → 75 5 → 25
trum with the Reference Spectrum: both spectra exhibit simi- 30 – 45 75 → 0 25 → 100
lar intensities of absorption at the same wavelengths. 45 – 55 0 100
Cefaclor as directed in the potassium bromide disk method Flow rate: 1.0 mL per minute.
under Infrared Spectrophotometry <2.25>, and compare the Time span of measurement: About 2.5 times as long as the
spectrum with the Reference Spectrum: both spectra exhibit retention time of cefaclor, beginning after the solvent peak.
similar intensities of absorption at the same wave numbers. System suitability—
(3) Dissolve 40 mg of Cefaclor in 0.5 mL of heavy water Test for required detectability: Measure exactly 1 mL of
for nuclear magnetic resonance spectroscopy and 1 drop of the standard solution, and add sodium dihydrogen phos-
deuterated hydrochloric acid for nuclear magnetic resonance phate TS, pH 2.5 to make exactly 20 mL. Confirm that the
spectroscopy, and determine the 1H spectrum of this solution peak area of cefaclor obtained from 20 mL of this solution is
as directed under Nuclear Magnetic Resonance Spectroscopy equivalent to 4 to 6z of that obtained from 20 mL of the
<2.21>, using sodium 3-trimethylsilylpropanesulfonate for standard solution.
nuclear magnetic resonance spectroscopy as an internal ref- System performance: When the procedure is run with 20
erence compound: it exhibits an AB type quartet signal A at mL of the standard solution under the above operating con-
around d 3.7 ppm, and a single signal or a sharp multiple ditions, the number of theoretical plates and the symmetry
signal B at around d 7.6 ppm. The ratio of the integrated in- factor of the peak of cefaclor are not less than 40,000 and
tensity of each signal, A:B, is about 2:5. 0.8 to 1.3, respectively.
(4) Perform the test with Cefaclor as directed under System repeatability: When the test is repeated 3 times
Flame Coloration Test <1.04> (2): a green color appears. with 20 mL of the standard solution under the above operat-
D : ＋105 – ＋1209(0.1 g calcu-
ing conditions, the relative standard deviations of the peak
lated on the anhydrous basis, water, 25 mL, 100 mm). areas and the retention times of cefaclor are not more than
2.0z, respectively.
Cefaclor according to Method 2, and perform the test. Pre- Water <2.48> Not more than 6.5z (0.2 g, volumetric titra-
pare the control solution with 2.0 mL of Standard Lead So- tion, back titration).
lution (not more than 20 ppm). Assay Weigh accurately an amount of Cefaclor and
(2) Arsenic <1.11>—Prepare the test solution by suspend- Cefaclor RS, equivalent to about 50 mg (potency), and dis-
ing 1.0 g of Cefaclor in 10 mL of N, N-dimethylformamide, solve each in 0.1 mol/L phosphate buffer solution (pH 4.5)
and perform the test (not more than 2 ppm).
JP XVII Official Monographs / Cefaclor Capsules 593
to make exactly 50 mL. Pipet 10 mL each of these solutions, acetonitrile, water, ethyl acetate and formic acid (30:10:10:1)
add exactly 10 mL of the internal standard solution, add 0.1 to a distance of about 10 cm, and air-dry the plate. Examine
mol/L phosphate buffer solution (pH 4.5) to make 50 mL, under ultraviolet light (main wavelength: 254 nm): the prin-
and use these solutions as the sample solution and the stand- cipal spot from the sample solution and the spot from the
ard solution, respectively. Perform the test with 10 mL each standard solution show the same R f value.
Purity Related substances—Weigh accurately not less than
5 Cefaclor Capsules, open the capsules and carefully take
out the contents, mix well, and powder, if necessary. Wash
the peak area of cefaclor to that of the internal standard.
the empty capsules with a little amount of diethyl ether, if
Amount [ mg (potency)] of cefaclor (C15H14ClN3O4S) necessary, allow the capsules to stand at room temperature
＝ MS × QT/QS × 1000 to vaporize adhering diethyl ether, and weigh accurately the
capsules to calculate the mass of the contents. Weigh accu-
MS: Amount [mg (potency)] of Cefaclor RS taken
rately a quantity of the contents, equivalent to about 0.25 g
Internal standard solution—A solution of 4-aminoacetophe- (potency) of Cefaclor, shake with 40 mL of 0.1 mol/L phos-
none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in phate buffer solution (pH 4.5) for 10 minutes, add the same
700). buffer solution to make exactly 50 mL, and filter through a
Operating conditions— 0.45-mm pore-size membrane filter. Discard the first 1 mL of
Detector: An ultraviolet absorption photometer (wave- the filtrate, and use the subsequent filtrate as the sample so-
length: 254 nm). lution. Separately, weigh accurately an amount of Cefaclor
Column: A stainless steel column 4.6 mm in inside diame- RS, equivalent to about 20 mg (potency), and dissolve in 0.1
ter and 15 cm in length, packed with octadecylsilanized silica mol/L phosphate buffer solution (pH 4.5) to make exactly
gel for liquid chromatography (5 mm in particle diameter). 20 mL. Pipet 2.5 mL of this solution, add the same buffer
Column temperature: A constant temperature of about solution to make exactly 50 mL, and use this solution as the
259 C. standard solution. Perform the test with exactly 20 mL each
Mobile phase: Dissolve 6.8 g of potassium dihydrogen of the sample solution and standard solution as directed
phosphate in 1000 mL of water, and adjust the pH to 3.4 under Liquid Chromatography <2.01> according to the fol-
with diluted phosphoric acid (3 in 500). To 940 mL of this lowing conditions, and determine the peak areas by the auto-
solution add 60 mL of acetonitrile. matic integration method. Calculate the amount of each
Flow rate: Adjust so that the retention time of cefaclor is related substance by the following equation: the amount of
about 7 minutes. each related substance is not more than 0.5z, and the total
System suitability— amount of the related substances is not more than 2.5z. If
System performance: When the procedure is run with 10 necessary, correct the fluctuation of the base line by per-
mL of the standard solution under the above operating con- forming the test in the same way with 20 mL of 0.1 mol/L
ditions, cefaclor and the internal standard are eluted in this phosphate buffer solution (pH 4.5).
Amount (z) of each related substance
than 5.
＝ MS/MT × ATi/AS × MM/C × 25/2
with 10 mL of the standard solution under the above operat- Total amount (z) of the related substances
ing conditions, the relative standard deviation of the ratios ＝ MS/MT × SATn/AS × MM/C × 25/2
of the peak area of cefaclor to that of the internal standard is
not more than 1.0z.
MT: Amount (mg) of the contents of Cefaclor Capsules
Containers and storage Containers—Tight containers. taken
Storage—Light-resistant. MM: Average mass (mg) of the contents in 1 capsule
ATi: Area of each peak other than cefaclor and solvent
from the sample solution
Cefaclor Capsules AS: Peak area of cefaclor from the standard solution
C: Labeled potency [mg (potency)] of Cefaclor in 1 cap-
セファクロルカプセル sule
Cefaclor Capsules contain not less than 90.0z and Proceed as directed in the operating conditions in the
not more than 110.0z of the labeled potency of Purity (3) under Cefaclor.
cefaclor (C15H14ClN3O4S: 367.81). System suitability—
solution, and add 0.1 mol/L phosphate buffer solution (pH
sules, with Cefaclor.
4.5) to make exactly 20 mL. Confirm that the peak area of
Identification Shake vigorously a quantity of the contents cefaclor obtained with 20 mL of this solution is equivalent to
of Cefaclor Capsules, equivalent to 20 mg (potency) of 3.5 to 6.5z of that obtained with 20 mL of the standard so-
Cefaclor, with 10 mL of water, centrifuge, and use the su- lution.
pernatant liquid as the sample solution. Separately, dissolve System performance: When the procedure is run with 20
20 mg of Cefaclor RS in 10 mL of water, and use this solu- mL of the standard solution under the above operating con-
tion as the standard solution. Perform the test with these so- ditions, the number of theoretical plates and the symmetry
lutions as directed under Thin-layer Chromatography <2.03>. factor of the peak of cefaclor are not less than 40,000 and
Spot 2 mL each of the sample solution and standard solution 0.8 to 1.3, respectively.
on a plate of silica gel with fluorescent indicator for thin- System repeatability: When the test is repeated 3 times
layer chromatography, develop the plate with a mixture of with 20 mL of the standard solution under the above operat-
594 Cefaclor Combination Granules / Official Monographs JP XVII
area of cefaclor is not more than 2.0z. Cefaclor Combination Granules
セファクロル複合顆粒
tion, back titration).
Cefaclor Combination Granules contain gastric-
soluble granules and enteric-soluble granules in one
Dissolution <6.10> When the test is performed at 50 revolu- package.
tions per minute according to the Paddle method, using 900 It contains cefaclor (C15H14ClN3O4S: 367.81)
mL of water as the dissolution medium, the dissolution rate equivalent to not less than 90.0z and not more than
in 15 minutes of Cefaclor Capsules is not less than 80z. 110.0z of the labeled total potency and the labeled
Start the test with 1 capsule of Cefaclor Capsules, with- potency of gastric-soluble granule, respectively.
ules, with Cefaclor, and divide into single-dose packages.
10 mL of the filtrate, pipet V mL of the subsequent filtrate, Identification Shake vigorously a quantity of Cefaclor
add water to make exactly V? mL so that each mL contains Combination Granules, equivalent to 20 mg (potency) of
about 20 mg (potency) of Cefaclor, and use this solution as Cefaclor according to the labeled total potency, with 10 mL
the sample solution. Separately, weigh accurately about 20 of water, centrifuge, and use the supernatant liquid as the
mg (potency) of Cefaclor RS, and dissolve in water to make sample solution. Separately, dissolve 20 mg of Cefaclor RS
exactly 20 mL. Pipet 1 mL of this solution, add water to in 10 mL of water, and use this solution as the standard solu-
make exactly 50 mL, and use this solution as the standard tion. Perform the test with these solutions as directed under
solution. Determine the absorbances, AT and AS, at 265 nm Thin-layer Chromatography <2.03>. Spot 2 mL each of the
of the sample solution and standard solution as directed sample solution and standard solution on a plate of silica gel
under Ultraviolet-visible Spectrophotometry <2.24>. with fluorescent indicator for thin-layer chromatography,
develop the plate with a mixture of acetonitrile, water, ethyl
acetate and formic acid (30:10:10:1) to a distance of about
of cefaclor (C15H14ClN3O4S)
10 cm, and air-dry the plate. Examine under ultraviolet light
＝ MS × AT/AS × V?/V × 1/C × 90
(main wavelength: 254 nm): the principal spot from the sam-
MS: Amount [mg (potency)] of Cefaclor RS taken ple solution and the spot from the standard solution show
C: Labeled amount [mg (potency)] of Cefaclor in 1 cap- the same R f value.
sule
Purity Related substances—Take out the total contents of
Assay Weigh accurately not less than 5 Cefaclor Capsules, not less than 5 packages of Cefaclor Combination Granules,
open the capsules and carefully take out the contents, mix add a small amount of 0.1 mol/L phosphate buffer solution
well, and powder, if necessary. Wash the empty capsules (pH 4.5), grind, add 0.1 mol/L phosphate buffer solution
with a little amount of diethyl ether, if necessary, allow the (pH 4.5), shake vigorously for 10 minutes, and add 0.1
capsules to stand at room temperature to vaporize adhering mol/L phosphate buffer solution (pH 4.5) to make exactly V
diethyl ether, and weigh accurately the capsules to calculate mL so that each mL contains about 5 mg (potency) of
the mass of the contents. Weigh accurately a quantity of the Cefaclor according to the labeled total potency. Pipet 10 mL
contents, equivalent to about 0.1 g (potency) of Cefaclor, of this solution, add 0.1 mol/L phosphate buffer solution
shake vigorously with 60 mL of 0.1 mol/L phosphate buffer (pH 4.5) to make exactly 25 mL, and filter through a mem-
solution (pH 4.5) for 10 minutes, add the same buffer solu- brane filter with a pore size not exceeding 0.45 mm. Discard
tion to make exactly 100 mL, and centrifuge. Pipet 10 mL of the first 1 mL of the filtrate, and use the subsequent filtrate
the supernatant liquid, add exactly 10 mL of the internal as the sample solution. Separately, weigh accurately an
standard solution and the same buffer solution to make 50 amount of Cefaclor RS, equivalent to about 20 mg (po-
mL, and use this solution as the sample solution. Separately, tency), and dissolve in 0.1 mol/L phosphate buffer solution
weigh accurately an amount of Cefaclor RS, equivalent to (pH 4.5) to make exactly 20 mL. Pipet 2 mL of this solution,
about 50 mg (potency), and dissolve in 0.1 mol/L phosphate add 0.1 mol/L phosphate buffer solution (pH 4.5) to make
buffer solution (pH 4.5) to make exactly 50 mL. Pipet 10 mL exactly 100 mL, and use this solution as the standard solu-
of this solution, add exactly 10 mL of the internal standard tion. Perform the test with exactly 50 mL each of the sample
solution and the same buffer solution to make 50 mL, and solution and standard solution as directed under Liquid
use this solution as the standard solution. Proceed as di- Chromatography <2.01> according to the following condi-
rected in the Assay under Cefaclor. tions, and determine each peak area in each solution by the
automatic integration method. Calculate the amount of each
Amount [mg (potency)] of cefaclor (C15H14ClN3O4S)
related substance by the following equation: the amount of
＝ M S × Q T / QS × 2
each related substance is not more than 0.6z, and the total
MS: Amount [mg (potency)] of Cefaclor RS taken amount of the related substances is not more than 2.8z. If
necessary, correct the fluctuation of the base line by per-
Internal standard solution—A solution of 4-aminoacetophe-
forming the test in the same way with 50 mL of 0.1 mol/L
none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in
phosphate buffer solution (pH 4.5).
700).
＝ MS × AT/AS × V/4 × {1/(C × T )}
Total amount (z) of the related substances
＝ MS × SAT/AS × V/4 × {1/(C × T )}
JP XVII Official Monographs / Cefaclor Combination Granules 595
MS: Amount [mg (potency)] of Cefaclor RS taken ard solution and the same buffer solution to make 50 mL,
AT: Area of each peak other than cefaclor, solvent and and use this solution as the sample solution. Separately,
excipient from the sample solution weigh accurately an amount of Cefaclor RS, equivalent to
SAT: Total area of the peaks other than cefaclor, solvent about 50 mg (potency), and dissolve in 0.1 mol/L phosphate
and excipient from the sample solution buffer solution (pH 4.5) to make exactly 50 mL. Pipet 10 mL
AS: Peak area of cefaclor from the standard solution of this solution, add exactly 10 mL of the internal standard
C: Labeled total potency [mg (potency)] of Cefaclor in 1 solution and the same buffer solution to make 50 mL, and
package use this solution as the standard solution. Then, proceed as
T: Number (pack) of Cefaclor Combination Granules directed in the Assay under Cefaclor.
Operating conditions— Amount [mg (potency)] of cefaclor (C15H14ClN3O4S)
Proceed as directed in the operating conditions in the ＝ MS × QT/QS × V/35
Purity (3) under Cefaclor.
Test for required detectability: Pipet 1 mL of standard so- Internal standard solution—A solution of 4-aminoacetophe-
lution, and add 0.1 mol/L phosphate buffer solution (pH none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in
4.5) to make exactly 20 mL. Confirm that the peak area of 700).
cefaclor obtained from 50 mL of this solution is equivalent to
3.5 to 6.5z of that obtained from 50 mL of the standard so-
lution.
mL of 1st fluid for dissolution test as the dissolution me-
dium, the dissolution rate in 60 minutes of Cefaclor Combi-
nation Granules is between 35z and 45z.
Start the test with the total content of 1 package of
factor of the peak of cefaclor are not less than 40,000 and
Cefaclor Combination Granules, withdraw not less than
between 0.8 and 1.3, respectively.
about 20 mL of the medium at the specified minute after
starting the test, and filter through a membrane filter with a
the filtrate, pipet V mL of the subsequent filtrate, add the
area of cefaclor is not more than 2.0z.
dissolution medium to make exactly V? mL so that each mL
Water <2.48> Not more than 5.5z (0.3 g, volumetric titra- contains about 20 mg (potency) of Cefaclor according to the
tion, back titration). labeled potency of gastric-soluble granule, and use this solu-
tion as the sample solution. Separately, weigh accurately an
amount of Cefaclor RS, equivalent to about 20 mg (po-
tency), and dissolve in the dissolution medium to make ex-
actly 20 mL. Pipet 2 mL of this solution, add the dissolution
(1) Total potency—Take out the total contents of 1
Cefaclor Combination Granules, add a little amount of 0.1
mol/L phosphate buffer solution (pH 4.5), grind well, add
265 nm of the sample solution and standard solution as di-
the same buffer solutions to make exactly V mL so that each
rected under Ultraviolet-visible Spectrophotometry <2.24>.
mL contains about 3.8 mg (potency) of Cefaclor according
to the labeled total potency after shaking vigorously for 10 Dissolution rate (z) of cefaclor (C15H14ClN3O4S) with
minutes, and centrifuge. Pipet 3 mL of the supernatant liq- respect to the labeled potency
uid, add exactly 10 mL of the internal standard solution and ＝ MS × AT/AS × V?/V × 1/C × 90
the same buffer solution to make 50 mL, and use this solu-
C: Labeled total potency [mg (potency)] of Cefaclor in 1
amount of Cefaclor RS, equivalent to about 50 mg (po-
pack
(pH 4.5) to make exactly 50 mL. Pipet 10 mL of this solu- Separately, when the test is performed at 50 revolutions
tion, add exactly 10 mL of the internal standard solution and per minute according to the Paddle method, using 900 mL of
the same buffer solution to make 50 mL, and use this solu- 2nd fluid for dissolution test as the dissolution medium, the
tion as the standard solution. Then, proceed as directed in dissolution rate in 60 minutes of Cefaclor Combination
the Assay under Cefaclor. Granules is not less than 70z.
Start the test with the total content of 1 package of
Cefaclor Combination Granules, withdraw not less than
＝ MS × QT/QS × V/15
about 20 mL of the medium at the specified minute after
MS: Amount [mg (potency)] of Cefaclor RS taken starting the test, and filter through a membrane filter with a
the filtrate, pipet V mL of the subsequent filtrate, add 0.01
mol/L hydrochloric acid TS to make exactly V? mL so that
700).
each mL contains about 20 mg (potency) of Cefaclor accord-
(2) Potency of gastric-soluble granule—Take out the
ing to the labeled total potency, and use this solution as the
total contents of 1 Cefaclor Combination Granules stir
sample solution. Separately, weigh accurately an amount of
gently with 60 mL of 0.1 mol/L phosphate buffer solution
Cefaclor RS, equivalent to about 20 mg (potency), dissolve
(pH 4.5) for 5 minutes, add the same buffer solution to
in the dissolution medium to make exactly 100 mL, and
warm at 379C for 60 minutes. Pipet 2 mL of this solution,
(potency) of Cefaclor according to the labeled potency of
add 0.01 mol/L hydrochloric acid TS to make exactly 20
gastric-soluble granule, and centrifuge. Pipet 7 mL of the
supernatant liquid, add exactly 10 mL of the internal stand-
596 Cefaclor Fine Granules / Official Monographs JP XVII
mine the absorbances, AT and AS, at 265 nm of the sample
solution and standard solution as directed under Ultraviolet- Cefaclor Fine Granules
visible Spectrophotometry <2.24>, using 0.01 mol/L hydro-
chloric acid TS as the blank. セファクロル細粒
Dissolution rate (z) of cefaclor (C15H14ClN3O4S) with
respect to the labeled potency Cefaclor Fine Granules contain not less than 90.0z
＝ MS × AT/AS × V?/V × 1/C × 90 and not more than 110.0z of the labeled potency of
cefaclor (C15H14ClN3O4S: 367.81).
C: Labeled total potency [mg (potency)] of Cefaclor in 1 Method of preparation Prepare as directed under Gran-
package ules, with Cefaclor.
Assay (1) Total potency—Take out the total contents of Identification Shake vigorously a quantity of Cefaclor Fine
not less than 5 Cefaclor Combination Granules, add a small Granules, equivalent to 20 mg (potency) of Cefaclor, with 10
amount of 0.1 mol/L phosphate buffer solution (pH 4.5), mL of water, centrifuge, and use the supernatant liquid as
grind well, add the same buffer solution so that each mL the sample solution. Separately, dissolve 20 mg (potency) of
containing about 5 mg (potency) of Cefaclor according to Cefaclor RS in 10 mL of water, and use this solution as the
the labeled total potency after shaking vigorously for 10 standard solution. Perform the test with these solutions as
minutes, and centrifuge. Pipet 2 mL of the supernatant liq- directed under Thin-layer Chromatography <2.03>. Spot 2
uid, add exactly 10 mL of the internal standard solution and mL each of the sample solution and standard solution on a
the same buffer solution to make 50 mL, and use this solu- plate of silica gel with fluorescent indicator for thin-layer
tion as the sample solution. Separately, weigh accurately an chromatography, develop the plate with a mixture of aceto-
amount of Cefaclor RS, equivalent to about 50 mg (po- nitrile, water, ethyl acetate and formic acid (30:10:10:1) to a
tency), and dissolve in 0.1 mol/L phosphate buffer solution distance of about 10 cm, and air-dry the plate. Examine
(pH 4.5) to make exactly 50 mL. Pipet 10 mL of this solu- under ultraviolet light (main wavelength: 254 nm): the prin-
tion, add exactly 10 mL of the internal standard solution and cipal spot from the sample solution and the spot from the
the same buffer solution to make 50 mL, and use this solu- standard solution show the same R f value.
tion as the standard solution. Then, proceed as directed in
Purity Related substances—Weigh accurately a quantity of
the Assay under Cefaclor.
Cefaclor Fine Granules after powdered if necessary, equiva-
Amount [mg (potency)] of cefaclor (C15H14ClN3O4S) lent to about 0.1 g (potency) of Cefaclor, shake with 40 mL
＝ MS × QT/QS × 1/5 of 0.1 mol/L phosphate buffer solution (pH 4.5) for 10
minutes, add the same buffer solution to make exactly 50
mL, and filter through a 0.45-mm pore-size membrane filter.
Internal standard solution—A solution of 4-aminoacetophe- Discard the first 1 mL of the filtrate, and use the subsequent
none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in filtrate as the sample solution. Separately, weigh accurately
700). an amount of Cefaclor RS, equivalent to about 20 mg (po-
(2) Potency of gastric-soluble granule—Stir gentry the tency), and dissolve in 0.1 mol/L phosphate buffer solution
total contents of not less than 5 Cefaclor Combination Gran- (pH 4.5) to make exactly 20 mL. Pipet 2 mL of this solution,
ules with about 100 mL of 0.1 mol/L phosphate buffer solu- add the same buffer solution to make exactly 100 mL, and
tion (pH 4.5) for 5 minutes, add the same buffer solution so use this solution as the standard solution. Perform the test
that each mL containing about 2 mg (potency) of Cefaclor with exactly 50 mL each of the sample solution and standard
according to the labeled potency of gastric-soluble granule, solution as directed under Liquid Chromatography <2.01>
and centrifuge. Pipet 5 mL of the supernatant liquid, add ex- according to the following conditions, and determine the
actly 10 mL of the internal standard solution and the same peak areas by the automatic integration method. Calculate
buffer solution to make 50 mL, and use this solution as the the amount of each related substance by the following equa-
sample solution. Separately, weigh accurately an amount of tion: the amount of each related substance is not more than
Cefaclor RS, equivalent to about 50 mg (potency), and dis- 0.5z, and the total amount of the related substances is not
solve in 0.1 mol/L phosphate buffer solution (pH 4.5) to more than 3.0z. If necessary, correct the fluctuation of the
make exactly 50 mL. Pipet 10 mL of this solution, add ex- base line by performing the test in the same way with 50 mL
actly 10 mL of the internal standard solution and the same of 0.1 mol/L phosphate buffer solution (pH 4.5).
buffer solution to make 50 mL, and use this solution as the
standard solution. Then, proceed as directed in the Assay
＝ MS/MT × AT/AS × 1/C × 5
under Cefaclor.
Total amount (z) of the related substances
＝ MS/MT × SAT/AS × 1/C × 5
＝ MS × QT/QS × 1/5
MT: Amount (g) of Cefaclor Fine Granules taken
Internal standard solution—A solution of 4-aminoacetophe- AT: Area of the peak other than cefaclor and the solvent
none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in from the sample solution
700). AS: Peak area of cefaclor from the standard solution
C: Labeled potency [mg (potency)] of cefaclor
(C15H14ClN3O4S) in 1 g
Purity (3) under Cefaclor.
JP XVII Official Monographs / Cefadroxil 597
System suitability— directed in the Assay under Cefaclor.

＝ M S × QT / QS × 2
4.5) to make exactly 20 mL. Confirm that the peak area of
cefaclor obtained with 50 mL of this solution is equivalent to MS: Amount [mg (potency)] of Cefaclor RS taken
3.5 to 6.5z of that obtained with 50 mL of the standard so-
lution.
700).
ditions, the number of theoretical plates and the symmetry Containers and storage Containers—Tight containers.
factor of the peak of cefaclor are not less than 40,000 and Storage—Light-resistant.
between 0.8 and 1.3, respectively.
with 50 mL of the standard solution under the above operat- Cefadroxil
area of cefaclor is not more than 2.0z. セファドロキシル
dose packages meet the requirement of the Mass variation
test.
C16H17N3O5S: 363.39
(6R,7R)-7-[(2R)-2-Amino-2-(4-
hydroxyphenyl)acetylamino]-3-methyl-8-oxo-5-thia-1-
in 15 minutes of Cefaclor Fine Granules is not less than
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
85z.
[50370-12-2]
Cefaclor Fine Granules, equivalent to about 0.25 g (potency)
Cefadroxil contains not less than 950 mg (potency)
of Cefaclor, withdraw not less than about 20 mL of the me-
and not more than 1020 mg (potency) per mg,
dium at the specified minute after starting the test, and filter
through a membrane filter with a pore size not exceeding 0.5
Cefadroxil is expressed as mass (potency) of cefadroxil
mm. Discard the first 10 mL of the filtrate, pipet V mL of the
(C16H17N3O5S).
each mL contains about 20 mg (potency) of Cefaclor, and use Description Cefadroxil occurs as a white to light yellow-
this solution as the sample solution. Separately, weigh accu- white powder.
rately about 20 mg (potency) of Cefaclor RS, and dissolve in It is sparingly soluble in water, slightly soluble in metha-
water to make exactly 20 mL. Pipet 1 mL of this solution, nol, and very slightly soluble in ethanol (95).
the standard solution. Determine the absorbances, AT and
solution of Cefadroxil (1 in 50,000) as directed under Ultra-
violet-visible Spectrophotometry <2.24>, and compare the
spectrum with the Reference Spectrum or the spectrum of a
<2.24>.
solution of Cefadroxil RS prepared in the same manner as
Dissolution rate (z) with respect to the labeled amount the sample solution: both spectra exhibit similar intensities
of cefaclor (C15H14ClN3O4S) of absorption at the same wavelengths.
＝ MS/MT × AT/AS × V?/V × 1/C × 90 (2) Determine the infrared absorption spectrum of
Cefadroxil as directed in the potassium bromide disk method
MT: Amount [mg (potency)] of Cefaclor Fine Granules
taken
Cefadroxil RS: both spectra exhibit similar intensities of ab-
C: Labeled amount [mg (potency)] of cefaclor
(C15H14ClN3O4S) in 1 g
Assay Weigh accurately a quantity of Cefaclor Fine Gran- Cefadroxil in a mixture of heavy water for nuclear magnetic
ules after powdered if necessary, equivalent to about 0.1 g resonance spectroscopy and deuterated hydrochloric acid
(potency) of Cefaclor, shake vigorously with 60 mL of 0.1 (3:1) (1 in 10), using sodium 3-(trimethylsilyl)propionate-d4
mol/L phosphate buffer solution (pH 4.5) for 10 minutes, for nuclear magnetic resonance spectroscopy as an internal
add the same buffer solution to make exactly 100 mL, and reference compound, as directed under Nuclear Magnetic
centrifuge. Pipet 10 mL of the supernatant liquid, add ex- Resonance Spectroscopy <2.21>: it exhibits a single signal A
actly 10 mL of the internal standard solution and the same at around d 2.1 ppm, a double signal B at around d 7.0 ppm,
buffer solution to make 50 mL, and use this solution as the and a double signal C at around d 7.5 ppm. The ratio of in-
sample solution. Separately, weigh accurately about 50 mg tegrated intensity of each signal, A:B:C, is about 3:2:2.
(potency) of Cefaclor RS, and dissolve in 0.1 mol/L phos-
D : ＋164 – ＋1829(0.6 g calcu-
phate buffer solution (pH 4.5) to make exactly 50 mL. Pipet
10 mL of this solution, add exactly 10 mL of the internal
standard solution and the same buffer solution to make 50 pH <2.54> Dissolve 1.0 g of Cefadroxil in 200 mL of water:
mL, and use this solution as standard solution. Proceed as pH of the solution is between 4.0 and 6.0.
598 Cefadroxil Capsules / Official Monographs JP XVII
Cefadroxil according to Method 2, and perform the test. Cefadroxil Capsules
Solution (not more than 20 ppm). セファドロキシルカプセル
(2) Related substances—Dissolve 0.1 g of Cefadroxil in 4
mL of a mixture of ethanol (99.5), water and diluted hydro-
Cefadroxil Capsules contain not less than 95.0z
chloric acid (1 in 5) (75:22:3), and use this solution as the
and not more than 105.0z of the labeled potency of
cefadroxil (C16H17N3O5S: 363.39).
mixture of ethanol (99.5), water and diluted hydrochloric
acid (1 in 5) (75:22:3) to make exactly 100 mL, and use this Method of preparation Prepare as directed under Cap-
solution as the standard solution. Perform the test with these sules, with Cefadroxil.
Identification Dissolve the contents of Cefadroxil Cap-
sules, equivalent to 10 mg (potency) of Cefadroxil, in 500
mL of water, and filter. Determine the absorption spectrum
phy. Develop with a mixture of ethyl acetate, water, ethanol
of the filtrate as directed under Ultraviolet-visible Spectro-
(99.5) and formic acid (14:5:5:1) to a distance of about 12
cm, and air-dry the plate. Spray evenly ninhydrin-citric acid-
acetic acid TS on the plate, and heat at 1009 C for 10
minutes: the spots other than the principal spot from the Water <2.48> Not more than 7.0z (0.15 g, volumetric
sample solution are not more intense than the spot from the titration, direct titration).
standard solution.
Water <2.48> Not less than 4.2z and not more than 6.0z ing to the following method: it meets the requirement of the
(0.5 g, volumetric titration, direct titration). Content uniformity test.
Place 1 capsule of Cefadroxil Capsules in 300 mL of
Assay Weigh accurately an amount of Cefadroxil and
water, disperse with the aid of ultrasonic waves, shake for 30
Cefadroxil RS, equivalent to about 50 mg (potency), dissolve
minutes, and add water to make exactly 500 mL. Pipet 5 mL
each in water to make exactly 500 mL, and use these solu-
of this solution, and add water to make exactly V mL so that
tions as the sample solution and the standard solution, re-
each mL contains about 0.1 mg (potency) of Cefadroxil.
spectively. Perform the test with exactly 10 mL each of the
Filter the solution, discard the first 10 mL of the filtrate, and
use the subsequent filtrate as the sample solution. Sepa-
rately, weigh accurately about 20 mg (potency) of Cefadroxil
conditions, and determine the peak areas, AT and AS, of
RS, dissolve in water to make exactly 200 mL, and use this
cefadroxil in each solution.
solution as the standard solution. Then, proceed as directed
Amount [ mg (potency)] of cefadroxil (C16H17N3O5S) in the Assay under Cefadroxil.
＝ MS × AT/AS × 1000
Amount [mg (potency)] of cefadroxil (C16H17N3O5S)
MS: amount [mg (potency)] of Cefadroxil RS taken ＝ MS × AT/AS × V/2
Operating conditions— MS: Amount [mg (potency)] of Cefadroxil RS taken
length: 262 nm).
mL of 0.05 mol/L acetic acid-sodium acetate buffer solution
(pH 4.0) as the dissolution medium, the dissolution rate in 90
minutes of Cefadroxil Capsules is not less than 80z.
Start the test with 1 capsule of Cefadroxil Capsules, with-
409 C.
Mobile phase: A mixture of a solution of potassium dihy-
drogenphosphate (17 in 12,500) and methanol (17:3).
Flow rate: Adjust so that the retention time of cefadroxil
is about 5 minutes.
trate, add water to make exactly V? mL so that each mL con-
tains about 22 mg (potency) of Cefadroxil, and use this solu-
System performance: Dissolve about 5 mg (potency) of
Cefadroxil and about 10 mg (potency) of propylene glycol
about 22 mg (potency) of Cefadroxil RS, and add water to
cefatrizine in 50 mL of water. When the procedure is run
make exactly 100 mL. Pipet 5 mL of this solution, add water
with 10 mL of this solution under the above operating condi-
tions, cefadroxil and cefatrizine are eluted in this order with
the resolution between these peaks being not less than 4.
water as the blank.
areas of cefadroxil is not more than 1.0z. Dissolution rate (z) with respect to the labeled amount
of cefadroxil (C16H17N3O5S)
＝ MS × AT/AS × V?/V × 1/C × 90
MS: Amount [mg (potency)] of Cefadroxil RS taken
C: Labeled amount [mg (potency)] of cefadroxil in 1 cap-
sule
JP XVII Official Monographs / Cefalexin 599
Assay Take out the contents of 20 Cefadroxil Capsules, Dissolution rate (z) with respect to the labeled amount
and combine. Weigh accurately the mass of the combined of cefadroxil (C16H17N3O5S)
contents, and powder. Weigh accurately a portion of the ＝ MS/MT × AT/AS × 1/C × 450
powder, equivalent to about 50 mg (potency) of Cefadroxil,
add 300 mL of water, shake for 30 minutes, then add water
MT: Amount (g) of Cefadroxil for Syrup taken
to make exactly 500 mL, and filter. Discard the first 10 mL
C: Labeled amount [mg (potency)] of cefadroxil in 1 g
of the filtrate, and use the subsequent filtrate as the sample
solution. Separately, weigh accurately an amount of Cefa- Assay Weigh accurately an amount of powdered Cefa-
droxil RS, equivalent to about 20 mg (potency), dissolve in droxil for Syrup, equivalent to about 50 mg (potency) of
water to make exactly 200 mL, and use this solution as the Cefadroxil, dissolve in water to make exactly 500 mL, and
standard solution. Then, proceed as directed in the Assay use this solution as the sample solution. Separately, weigh
under Cefadroxil. accurately an amount of Cefadroxil RS, equivalent to about
20 mg (potency), dissolve in water to make exactly 200 mL,
and use this solution as the standard solution. Then, proceed
＝ MS × AT/AS × 5/2
as directed in the Assay under Cefadroxil.
Containers and storage Containers—Tight containers. ＝ MS × AT/AS × 5/2
Cefadroxil for Syrup Containers and storage Containers—Tight containers.
シロップ用セファドロキシル
Cefalexin
Cefadroxil for Syrup is a preparation for syrup, セファレキシン
which is suspended before use.
than 110.0z of the labeled potency of cefadroxil
(C16H17N3O5S: 363.39).
Method of preparation Prepare as directed under Prepara-
tions for Syrups, with Cefadroxil.
Identification Dissolve an amount of Cefadroxil for Syrup, C16H17N3O4S: 347.39
equivalent to 10 mg (potency) of Cefadroxil, in 500 mL of (6R,7R)-7-[(2R)-2-Amino-2-phenylacetylamino]-3-
water, and determine the absorption spectrum of this solu- methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
tion as directed under Ultraviolet-visible Spectrophotometry carboxylic acid
<2.24>: it exhibits maxima between 228 nm and 232 nm, and [15686-71-2]
Cefalexin contains not less than 950 mg (potency)
and not more than 1030 mg (potency) per mg, calcu-
lated on the anhydrous basis. The potency of Cefalex-
Uniformity of dosage units <6.02> The syrup in single-dose in is expressed as mass (potency) of cefalexin
packages meets the requirement of the Mass variation test. (C16H17N3O4S).
Dissolution <6.10> When the test is performed at 50 revolu- Description Cefalexin occurs as a white to light yellowish
tions per minute according to the Paddle method (put the white, crystals or crystalline powder.
sample in the dissolution medium so that it disperses), using It is sparingly soluble in water, slightly soluble in metha-
900 mL of water as the dissolution medium, the dissolution nol, and practically insoluble in ethanol (95) and in N, N-
rate in 15 minutes of Cefadroxil for Syrup is not less than dimethylformamide.
85z. It is hygroscopic.
Start the test with accurately weighed amount of
Cefadroxil for Syrup, equivalent to about 0.1 g (potency) of
solution of Cefalexin (3 in 100,000) as directed under Ultra-
Cefadroxil, withdraw not less than 20 mL of the medium at
Discard the first 10 mL of the filtrate, pipet 4 mL of the sub-
sequent filtrate, add water to make exactly 20 mL, and use
Cefalexin as directed in the potassium bromide disk method
rately about 22 mg (potency) of Cefadroxil RS, and dissolve
in water to make exactly 100 mL. Pipet 5 mL of this solu-
(3) Determine the 1H spectrum of a solution of Cefalexin
as the standard solution. Determine the absorbances, AT and
in heavy water for nuclear magnetic resonance spectroscopy
(1 in 200) as directed under Nuclear Magnetic Resonance
Spectroscopy <2.21>, using sodium 3-trimethylsilylpropane-
<2.24>.
sulfonate for nuclear magnetic resonance spectroscopy as an
internal reference compound: it exhibits a single signal A at
around d 1.8 ppm, and a single or a sharp multiple signal B
600 Cefalexin / Official Monographs JP XVII
at around d 7.5 ppm. The ratio of integrated intensity of equivalent to 1.8 to 2.2z of that obtained from 20 mL of the
these signals, A:B, is about 3:5. standard solution.
D : ＋144 – ＋1589(0.125 g calcu-
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of factor of the peak of cefalexin are not less than 150,000 and
Cefalexin according to Method 4, and perform the test. Pre- between 0.8 and 1.3, respectively.
pare the control solution with 2.0 mL of Standard Lead So- System repeatability: When the test is repeated 3 times
lution (not more than 10 ppm). with 20 mL of the standard solution under the above operat-
(2) Arsenic <1.11>—Prepare the test solution with 1.0 g ing conditions, the relative standard deviation of the reten-
of Cefalexin by suspending in 10 mL of N, N-dimethylfor- tion time and the peak areas of cefalexin are not more than
mamide, and perform the test (not more than 2 ppm). 2.0z, respectively.
(3) Related substances—Dissolve about 25 mg of
Cefalexin in a solution of potassium dihydrogenphosphate
(9 in 500) to make 5 mL, and use this solution as the sample
solution. Pipet 1 mL of the sample solution, add a solution Assay Weigh accurately an amount of Cefalexin and
of potassium dihydrogenphosphate (9 in 500) to make ex- Cefalexin RS, equivalent to about 25 mg (potency), dissolve
actly 100 mL, and use this solution as the standard solution. each in 0.1 mol/L phosphate buffer solution (pH 4.5) to
Perform the test with exactly 20 mL each of the sample solu- make exactly 25 mL. Pipet 10 mL of these solutions, add
tion and standard solution as directed under Liquid Chroma- exactly 5 mL of the internal standard solution, then add 0.1
tography <2.01> according to the following conditions, and mol/L phosphate buffer solution (pH 4.5) to make 50 mL,
determine the areas of each peak by the automatic integra- and use these solutions as the sample solution and standard
tion method. If necessary, correct the change of the base-line solution. Perform the test with 10 mL each of the sample so-
due to the potassium dihydrogenphosphate solution by lution and standard solution as directed under Liquid Chro-
proceeding in the same manner with 20 mL of a solution of matography <2.01> according to the following conditions,
potassium dihydrogenphosphate (9 in 500): each peak area and calculate the ratios, QT and QS, of the peak area of
other than cefalexin from the sample solution is not larger cefalexin to that of the internal standard.
than the peak area of cefalexin from the standard solution,
Amount [ mg (potency)] of cefalexin (C16H17N3O4S)
and the total area of the peaks other than cefalexin from the
＝ MS × QT/QS × 1000
sample solution which are larger than 1/50 times the peak
area of cefalexin from the standard solution is not larger MS: amount [mg (potency)] of Cefalexin RS taken
than 5 times of the peak area of cefalexin from the standard
Internal standard solution—A solution of m-hydroxy-
solution.
acetophenone in 0.1 mol/L phosphate buffer solution (pH
4.5) (1 in 1500).
length: 254 nm).
length: 254 nm).
259 C.
Mobile phase A: Dissolve 1.0 g of sodium 1-pentanesul-
259C.
fonate in 1000 mL of water, add 15 mL of triethylamine,
Mobile phase: Dissolve 6.8 g of potassium dihydrogen-
and adjust to pH 2.5 with phosphoric acid.
phosphate in 1000 mL of water, adjust to pH 3.0 with
Mobile phase B: Dissolve 1.0 g of sodium 1-pentanesul-
fonate in 300 mL of water, add 15 mL of triethylamine, and
adjust to pH 2.5 with phosphoric acid. To this solution add
Flow rate: Adjust so that the retention time of cefalexin is
350 mL of acetonitrile and 350 mL of methanol.
about 7 minutes.
Time after injection Mobile phase A Mobile phase B ditions, cefalexin and the internal standard are eluted in this
of sample (min) (volz) (volz) order with the resolution between these peaks being not less
than 6.
0–1 100 0
1 – 34.5 100 → 0 0 → 100
34.5 – 35.5 0 100
of the peak area of cefalexin to that of the internal standard
Flow rate: 1.0 mL per minute. is not more than 1.0z.
retention time of cefalexin, beginning after the solvent peak. Containers and storage Containers—Tight containers.
solution, add a solution of potassium dihydrogenphosphate
(9 in 500) to make exactly 100 mL. Confirm that the peak
area of cefalexin obtained from 20 mL of this solution is
JP XVII Official Monographs / Cefalexin Capsules 601

Cefalexin Capsules tions per minute according to the Paddle method, using 900
mL of water as the dissolution medium, the dissolution rates
セファレキシンカプセル in 30 minutes of 125-mg (potency) capsule and in 60 minutes
of 250-mg (potency) capsule are not less than 75z and 80z,
respectively.
Cefalexin Capsules contain not less than 93.0z and
Start the test with 1 capsule of Cefalexin Capsules, with-
not more than 107.0z of the labeled potency of
cefalexin (C16H17N3O4S: 347.39).
Method of preparation Prepare as directed under Cap- filter with a pore size not exceeding 0.5 mm. Discard the first
sules, with Cefalexin. 10 mL of the filtrate, pipet V mL of the subsequent filtrate,
Identification Take out the contents of Cefalexin Capsules,
about 22 mg (potency) of Cefalexin, and use this solution as
to a quantity of the contents, equivalent to 70 mg (potency)
the sample solution. Separately, weigh accurately an amount
of Cefalexin, add 25 mL of water, shake vigorously for 5
of Cefalexin RS, equivalent to about 22 mg (potency), and
minutes, and filter. To 1 mL of the filtrate add water to
dissolve in water to make exactly 50 mL. Pipet 5 mL of this
make 100 mL. Determine the absorption spectrum of this so-
lution as directed under Ultraviolet-visible Spectrophotome-
lution as the standard solution. Perform the test with the
try <2.24>: it exhibits a maximum between 260 nm and 264
sample solution and standard solution as directed under Ul-
nm.
traviolet-visible Spectrophotometry <2.24>, and determine
Water <2.48> Not more than 10.0z (0.2 g, volumetric the absorbances, AT and AS, at 262 nm.
titration, back titration).
Uniformity of dosage units <6.02> Perform the Mass varia- of cefalexin (C16H17N3O4S)
tion test, or the Content uniformity test according to the fol- ＝ MS × AT/AS × V?/V × 1/C × 90
MS: Amount [mg (potency)] of Cefalexin RS taken
Open 1 capsule of Cefalexin Capsules, add 3V/5 mL of
C: Labeled amount [mg (potency)] of cefalexin
0.1 mol/L phosphate buffer solution (pH 4.5), shake vigor-
(C16H17N3O4S) in 1 capsule
ously for 10 minutes, add 0.1 mol/L phosphate buffer solu-
tion (pH 4.5) to make exactly V mL so that each mL con- Assay Take out the contents of not less than 20 capsules of
tains about 1.25 mg (potency) of Cefalexin. Centrifuge this Cefalexin Capsules, weigh accurately the mass of the con-
solution, pipet 2 mL of the supernatant liquid, add exactly tents, and powder. Weigh accurately a portion of the pow-
10 mL of the internal standard solution, add 0.1 mol/L der, equivalent to about 0.1 g (potency) of Cefalexin, add 60
phosphate buffer solution (pH 4.5) to make 100 mL, and use mL of 0.1 mol/L phosphate buffer solution (pH 4.5), shake
this solution as the sample solution. Separately, weigh accu- vigorously for 10 minutes, add 0.1 mol/L phosphate buffer
rately an amount of Cefalexin RS, equivalent to about 25 mg solution (pH 4.5) to make exactly 100 mL, and centrifuge.
(potency), dissolve in 0.1 mol/L phosphate buffer solution Pipet 2 mL of the supernatant liquid, add exactly 10 mL of
(pH 4.5) to make exactly 100 mL. Pipet 10 mL of this solu- the internal standard solution, add 0.1 mol/L phosphate
tion, add exactly 10 mL of the internal standard solution, buffer solution (pH 4.5) to make 100 mL, and use this solu-
add 0.1 mol/L phosphate buffer solution (pH 4.5) to make tion as the sample solution. Separately, weigh accurately an
100 mL, and use this solution as the standard solution. Per- amount of Cefalexin RS, equivalent to about 20 mg (po-
form the test with 10 mL each of the sample solution and tency), dissolve in 0.1 mol/L phosphate buffer solution (pH
standard solution as directed under Liquid Chromatography 4.5) to make exactly 100 mL. Pipet 10 mL of this solution,
<2.01> according to the following conditions, and calculate add exactly 10 mL of the internal standard solution, add 0.1
the ratios, QT and QS, of the peak area of cefalexin to that of mol/L phosphate buffer solution (pH 4.5) to make 100 mL,
the internal standard. and use this solution as the standard solution. Perform the
test with 10 mL each of the sample solution and standard
Amount [mg (potency)] of cefalexin (C16H17N3O4S)
＝ MS × QT/QS × V/20
MS: Amount [mg (potency)] of Cefalexin RS taken ratios, QT and QS, of the peak area of cefalexin to that of the
internal standard.
cetophenone in 0.1 mol/L phosphate buffer solution (pH Amount [mg (potency)] of cefalexin (C16H17N3O4S)
4.5) (1 in 15,000). ＝ MS × QT/QS × 5
Assay. Internal standard solution—A solution of m-hydroxya-
System suitability— cetophenone in 0.1 mol/L phosphate buffer solution (pH
System performance: When the procedure is run with 10 4.5) (1 in 15,000).
mL of the standard solution under the above operating con- Operating conditions—
ditions, cefalexin and the internal standard are eluted in this Detector: An ultraviolet absorption photometer (wave-
order with the resolution between these peaks being not less length: 254 nm).
than 8. Column: A stainless steel column 3.0 mm in inside diame-
System repeatability: When the test is repeated 6 times ter and 7.5 cm in length, packed with octadecylsilanized
with 10 mL of the standard solution under the above operat- silica gel for liquid chromatography (3 mm in particle diame-
ing conditions, the relative standard deviation of the ratio of ter).
the peak area of cefalexin to that of the internal standard Column temperature: A constant temperature of about
substance is not more than 1.0z. 259C.
602 Cefalexin Combination Granules / Official Monographs JP XVII
Mobile phase: Dissolve 2.72 g of potassium dihydrogen Internal standard solution—A solution of m-hydroxya-
phosphate in 1000 mL of water, and adjust the pH to 3.0 cetophenone in 0.1 mol/L phosphate buffer solution (pH
with diluted phosphoric acid (3 in 500). To 800 mL of this 4.5) (1 in 15,000).
solution add 200 mL of methanol. (2) Potency of gastric-soluble granules—To the total
Flow rate: Adjust so that the retention time of cefalexin is amount of the content of 1 package of Cefalexin Combina-
about 6 minutes. tion Granules, add 3 V/5 mL of 0.1 mol/L phosphate buffer
System suitability— solution (pH 4.5), shake gently for 5 minutes, add 0.1 mol/L
System performance: When the procedure is run with 10 phosphate buffer solution (pH 4.5) to make exactly V mL so
mL of the standard solution under the above operating con- that each mL contains about 0.6 mg (potency) of Cefalexin
ditions, cefalexin and the internal standard are eluted in this according to the labeled potency of gastric-soluble granules,
order with the resolution between these peaks being not less and centrifuge. Pipet 7 mL of the supernatant liquid, add ex-
than 8. actly 20 mL of the internal standard solution, add 0.1 mol/L
System repeatability: When the test is repeated 6 times phosphate buffer solution (pH 4.5) to make 200 mL, and use
with 10 mL of the standard solution under the above operat- this solution as the sample solution. Then, proceed as di-
ing conditions, the relative standard deviation of the ratio of rected in the Assay (1) Total potency.
the peak area of cefalexin to that of the internal standard is
Amount [mg (potency)] of cefalexin (C16H17N3O4S)
not more than 1.0z.
＝ MS × QT/QS × V/35
Cefalexin Combination Granules cetophenone in 0.1 mol/L phosphate buffer solution (pH
4.5) (1 in 15,000).
セファレキシン複合顆粒
Cefalexin Combination Granules contain gastric- mL of 1st fluid for dissolution test as the dissolution me-
soluble granules and enteric-soluble granules in one dium, the dissolution rate in 30 minutes of Cefalexin Combi-
package. nation Granules is between 25z and 35z.
It contains not less than 90.0z and not more than Start the test with the total amount of the content of 1
110.0z of cefalexin (C16H17N3O4S: 347.39) for the package of Cefalexin Combination Granules, withdraw not
labeled total potency and the labeled potency of gas- less than about 20 mL of the medium at the specified minute
tric-soluble granules, respectively. after starting the test, and filter through a membrane filter
ules, with Cefalexin, and pack into single-dose packages.
the dissolution medium to make exactly V? mL so that each
Identification Powder Cefalexin Combination Granules, mL contains about 22 mg (potency) of Cefalexin according to
weigh a portion of the powder, equivalent to 30 mg (po- the labeled potency of gastric-soluble granules, and use this
tency) of Cefalexin according to the labeled total potency, solution as the sample solution. Separately, weigh accurately
shake vigorously for 5 minutes with 100 mL of water, and an amount of Cefalexin RS, equivalent to about 22 mg (po-
centrifuge. To 2 mL of the supernatant liquid add water to tency), and dissolve in the dissolution medium to make ex-
make 20 mL. Determine the absorption spectrum of this so- actly 50 mL. Pipet 5 mL of this solution, add the dissolution
lution as directed under Ultraviolet-visible Spectrophotome- medium to make exactly 100 mL, and use this solution as the
try <2.24>: it exhibits a maximum between 260 nm and 264 standard solution. Determine the absorbances, AT and AS, at
nm. 262 nm of the sample solution and standard solution as di-
tion, direct titration). Dissolution rate (z) of cefalexin (C16H17N3O4S)
with respect to the labeled potency
＝ MS × AT/AS × V?/V × 1/C × 90
Content uniformity test. MS: Amount [mg (potency)] of Cefalexin RS taken
(1) Total potency—To the total amount of the content of C: Labeled total potency [mg (potency)] of Cefalexin in 1
1 package of Cefalexin Combination Granules add a small package
amount of 0.1 mol/L phosphate buffer solution (pH 4.5),
When the test is performed at 50 revolutions per minute
grind, add 3 V/5 mL of 0.1 mol/L phosphate buffer solution
according to the Paddle method, using 900 mL of 2nd fluid
(pH 4.5), shake vigorously for 10 minutes, add 0.1 mol/L
for dissolution test as the dissolution medium, the dissolu-
phosphate buffer solution (pH 4.5) to make exactly V mL so
tion rate in 60 minutes of 200 mg (potency) preparation is
that each mL contains about 2 mg (potency) of Cefalexin
not less than 80z, and the dissolution rate in 45 minutes of
according to the labeled total potency, and centrifuge. Pipet
500 mg (potency) preparation is not less than 75z.
2 mL of the supernatant liquid, add exactly 20 mL of the
Start the test with the total amount of the content of 1
internal standard solution, add 0.1 mol/L phosphate buffer
package of Cefalexin Combination Granules, withdraw not
solution (pH 4.5) to make 200 mL, and use this solution as
less than about 20 mL of the medium at the specified minute
the sample solution. Then, proceed as directed in the Assay
(1) Total potency.
Amount [mg (potency)] of cefalexin (C16H17N3O4S) mL of the filtrate, pipet V mL of the subsequent filtrate, add
＝ MS × QT/QS × V/10 the dissolution medium to make exactly V? mL so that each
mL contains about 22 mg (potency) of Cefalexin according to
JP XVII Official Monographs / Cefalexin for Syrup 603
the labeled total potency, and use this solution as the sample System repeatability: When the test is repeated 6 times
solution. Separately, weigh accurately an amount of with 10 mL of the standard solution under the above operat-
Cefalexin RS, equivalent to about 22 mg (potency), dissolve ing conditions, the relative standard deviation of the ratio of
in the dissolution medium to make exactly 50 mL. Pipet 5 the peak area of cefalexin to that of the internal standard is
mL of this solution, add the dissolution medium to make not more than 1.0z.
exactly 100 mL, and use this solution as the standard solu- (2) Potency of gastric-soluble granules—Take out the
tion. Determine the absorbances, AT and AS, at 262 nm of content from not less than 20 packages of Cefalexin Combi-
the sample solution and standard solution as directed under nation Granules, weigh accurately a quantity, equivalent to
Ultraviolet-visible Spectrophotometry <2.24>. about 0.3 g (potency) of Cefalexin according to the labeled
potency of gastric-soluble granules, shake gently for 5
Dissolution rate (z) of cefalexin (C16H17N3O4S)
minutes with 200 mL of 0.1 mol/L phosphate buffer solu-
with respect to the labeled potency
tion (pH 4.5), add 0.1 mol/L phosphate buffer solution (pH
＝ MS × AT/AS × V?/V × 1/C × 90
4.5) to make exactly 300 mL, and centrifuge. Pipet 2 mL of
MS: Amount [mg (potency)] of Cefalexin RS taken the supernatant liquid, add exactly 10 mL of the internal
C: Labeled total potency [mg (potency)] of Cefalexin in 1 standard solution, and add 0.1 mol/L phosphate buffer so-
package lution (pH 4.5) to make 100 mL, and use this solution as the
sample solution. Then, proceed as directed in the Assay (1)
Assay (1) Total potency—Powder the total amount of the
Total potency.
content obtained from not less than 20 packages of Cefalex-
in Combination Granules, weigh accurately a portion of the Amount [mg (potency)] of cefalexin (C16H17N3O4S)
powder, equivalent to about 0.5 g (potency) of Cefalexin, ＝ MS × QT/QS × 15
shake vigorously for 10 minutes with 150 mL of 0.1 mol/L
phosphate buffer solution (pH 4.5), add 0.1 mol/L phos-
phate buffer solution (pH 4.5) to make exactly 250 mL, and Internal standard solution—A solution of m-hydroxya-
centrifuge. Pipet 2 mL of this solution, add exactly 20 mL of cetophenone in 0.1 mol/L phosphate buffer solution (pH
the internal standard solution, and add 0.1 mol/L phosphate 4.5) (1 in 15,000).
buffer solution (pH 4.5) to make 200 mL, and use this solu-
amount of Cefalexin RS, equivalent to about 20 mg (po-
(pH 4.5) to make exactly 100 mL. Pipet 10 mL of this solu- Cefalexin for Syrup
tion, add exactly 10 mL of the internal standard solution,
シロップ用セファレキシン
and add 0.1 mol/L phosphate buffer solution (pH 4.5) to
Perform the test with 10 mL each of the sample solution and Cefalexin for Syrup is a preparation for syrup,
standard solution as directed under Liquid Chromatography which is dissolved or suspended before use.
<2.01> according to the following conditions, and calculate It contains not less than 90.0z and not more
the ratios, QT and QS, of the peak area of cefalexin to that of than 110.0z of the labeled potency of cefalexin
the internal standard. (C16H17N3O4S: 347.39).
Amount [mg (potency)] of cefalexin (C16H17N3O4S) Method of preparation Prepare as directed under Prepara-
＝ MS × QT/QS × 25 tions for Syrups, with Cefalexin.
MS: Amount [mg (potency)] of Cefalexin RS taken Identification Dissolve a quantity of Cefalexin for Syrup,
equivalent to 3 mg (potency) of Cefalexin, in water to make
100 mL. Determine the absorption spectrum of this solution
cetophenone in 0.1 mol/L phosphate buffer solution (pH
4.5) (1 in 15,000).
Detector: An ultraviolet absorption photometer (wave- Water <2.48> Not more than 5.0z (0.4 g, volumetric titra-
length: 254 mn). tion, back titration).
ter and 7.5 cm in length, packed with octadecylsilanized
ing to the following method: Cefalexin for Syrup in single-
dose packages meets the requirement of the Content unifor-
ter).
mity test.
Take out the total contents of 1 package of Cefalexin for
259 C.
Syrup, add 3V/5 mL of 0.1 mol/L phosphate buffer solu-
tion (pH 4.5), shake vigorously for 10 minutes, add 0.1
phosphate in 1000 mL of water, and adjust to pH 3.0 with
mol/L phosphate buffer solution (pH 4.5) to make exactly
V mL so that each mL contains about 1 mg (potency) of
Cefalexin, and centrifuge. Pipet 2 mL of the supernatant
Flow rate: Adjust so that the retention time of cefalexin is
liquid, add exactly 10 mL of the internal standard solution,
about 6 minutes.
add 0.1 mol/L phosphate buffer solution (pH 4.5) to make
100 mL, and use this solution as the sample solution. Then,
proceed as directed in the Assay.
ditions, cefalexin and the internal standard are eluted in this Amount [mg (potency)] of cefalexin (C16H17N3O4S)
order with the resolution between these peaks being not less ＝ MS × QT/QS × V/20
than 8.
604 Cefalotin Sodium / Official Monographs JP XVII
MS: Amount [mg (potency)] of Cefalexin RS taken Column temperature: A constant temperature of about
259C.
cetophenone in 0.1 mol/L phosphate buffer solution (pH
phosphate in 1000 mL of water, and adjust the pH to 3.0
4.5) (1 in 15,000).
with diluted phosphoric acid (3 in 500). To 800 mL of this
Dissolution <6.10> When the test is performed at 50 revolu- solution add 200 mL of methanol.
tions per minute according to the Paddle method, using 900 Flow rate: Adjust so that the retention time of cefalexin is
mL of water as the dissolution medium, the dissolution rate about 6 minutes.
in 15 minutes of Cefalexin for Syrup is not less than 80z. System suitability—
Start the test with an accurately weighed amount of System performance: When the procedure is run with 10
Cefalexin, equivalent to about 0.25 g (potency) of Cefalexin mL of the standard solution under the above operating con-
for Syrup, withdraw not less than 20 mL of the medium at ditions, cefalexin and the internal standard are eluted in this
the specified minute after starting the test, and filter through order with the resolution between these peaks being not less
a membrane filter with a pore size not exceeding 0.5 mm. than 8.
Discard the first 10 mL of the filtrate, pipet 2 mL of the sub- System repeatability: When the test is repeated 6 times
sequent filtrate, add water to make exactly 25 mL, and use with 10 mL of the standard solution under the above operat-
this solution as the sample solution. Separately, weigh accu- ing conditions, the relative standard deviation of the ratio of
rately an amount of Cefalexin RS, equivalent to about 22 mg the peak area of cefalexin to that of the internal standard is
(potency), and dissolve in water to make exactly 50 mL. not more than 1.0z.
the test with the sample solution and standard solution as di-
and determine the absorbances, AT and AS, at 262 nm.
Cefalotin Sodium
of cefalexin (C16H17N3O4S) セファロチンナトリウム
＝ MS/MT × AT/AS × 1/C × 1125
MT: Amount (g) of Cefalexin for Syrup taken
C: Labeled amount [mg (potency)] of cefalexin
(C16H17N3O4S) in 1 g
Assay Powder Cefalexin for Syrup, if necessary, and weigh
C16H15N2NaO6S2: 418.42
accurately a portion of the powder, equivalent to about 0.1 g
Monosodium (6R,7R)-3-acetoxymethyl-8-oxo-7-[2-
(potency) of Cefalexin, add 60 mL of 0.1 mol/L phosphate
(thiophen-2-yl)acetylamino]-5-thia-1-azabicyclo[4.2.0]oct-
buffer solution (pH 4.5), shake vigorously for 10 minutes,
2-ene-2-carboxylate
add 0.1 mol/L phosphate buffer solution (pH 4.5) to make
[58-71-9]
exactly 100 mL, and centrifuge. Pipet 2 mL of the superna-
tant liquid, add exactly 10 mL of the internal standard solu-
Cefalotin Sodium contains not less than 920 mg
tion, add 0.1 mol/L phosphate buffer solution (pH 4.5) to
Separately, weigh accurately an amount of Cefalexin RS,
Cefalotin Sodium is expressed as mass (potency) of
equivalent to about 20 mg (potency), dissolve in 0.1 mol/L
cefalotin (C16H16N2O6S2: 396.44).
phosphate buffer solution (pH 4.5) to make exactly 100 mL.
Pipet 10 mL of this solution, add exactly 10 mL of the inter- Description Cefalotin Sodium occurs as white to light yel-
nal standard solution, add 0.1 mol/L phosphate buffer solu- lowish white, crystals or crystalline powder.
tion (pH 4.5) to make 100 mL, and use this solution as the It is freely soluble in water, slightly soluble in methanol,
standard solution. Perform the test with 10 mL each of the very slightly soluble in ethanol (95), and practically insoluble
sample solution and standard solution as directed under Liq- in acetonitrile.
solution of Cefalotin Sodium (1 in 50,000) as directed under
of cefalexin to that of the internal standard.
Amount [mg (potency)] of cefalexin (C16H17N3O4S) the spectrum with the Reference Spectrum or the spectrum
＝ MS × QT/QS × 5 of a solution of Cefalotin Sodium RS prepared in the same
Internal standard solution—A solution of m-hydroxya- (2) Determine the infrared absorption spectrum of
cetophenone in 0.1 mol/L phosphate buffer solution (pH Cefalotin Sodium as directed in the potassium bromide disk
4.5) (1 in 15,000). method under Infrared Spectrophotometry <2.25>, and com-
Operating conditions— pare the spectrum with the Reference Spectrum or the spec-
Detector: An ultraviolet absorption photometer (wave- trum of Cefalotin Sodium RS: both spectra exhibit similar
length: 254 nm). intensities of absorption at the same wave numbers.
Column: A stainless steel column 3.0 mm in inside diame- (3) Determine the 1H spectrum of a solution of Cefalotin
ter and 7.5 cm in length, packed with octadecylsilanized Sodium in heavy water for nuclear magnetic resonance
silica gel for liquid chromatography (3 mm in particle diame- spectroscopy (1 in 10) as directed under Nuclear Magnetic
ter). Resonance Spectroscopy <2.21>, using sodium 3-trimethyl-
JP XVII Official Monographs / Cefatrizine Propylene Glycolate 605
silylpropanesulfonate for nuclear magnetic resonance spec- Assay Weigh accurately an amount of Cefalotin Sodium
troscopy as an internal reference compound: it exhibits a and Cefalotin Sodium RS, equivalent to about 25 mg (po-
single signal A at around d 2.1 ppm, a single or sharp multi- tency), and dissolve each in the mobile phase to make exactly
ple signal B at around d 3.9 ppm, and a multiple signal C at 25 mL, and use these solutions as the sample solution and
around d 7.0 ppm. The ratio of the integrated intensity of standard solution. Perform the test with exactly 10 mL each
these signals, A:B:C, is about 3:2:2. of the sample solution and standard solution as directed
(4) Cefalotin Sodium responds to the Qualitative Tests under Liquid Chromatography <2.01> according to the fol-
<1.09> (1) for sodium salt. lowing conditions, and determine the peak areas, AT and AS,
of cefalotin in each solution.
D : ＋124 – ＋1349 (5 g, water,
100 mL, 100 mm). Amount [ mg (potency)] of cefalotin (C16H16N2O6S2)
＝ MS × AT/AS × 1000
1.0 g of Cefalotin Sodium in 10 mL of water is between 4.5 MS: Amount [mg (potency)] of Cefalotin Sodium RS
and 7.0. taken
Purity (1) Clarity and color of solution—Dissolve 1.0 g Operating conditions—
of Cefalotin Sodium in 10 mL of water: the solution is clear. Detector: An ultraviolet absorption photometer (wave-
The absorbance of this solution at 450 nm, determined as length: 254 nm).
directed under Ultraviolet-visible Spectrophotometry <2.24>, Column: A stainless steel column 4.6 mm in inside diame-
is not more than 0.20. ter and 25 cm in length, packed with octadecylsilanized silica
(2) Heavy metals <1.07>—Proceed with 1.0 g of Cefalo- gel for liquid chromatography (5 mm in particle diameter).
tin Sodium according to Method 2, and perform the test. Column temperature: A constant temperature of about
Prepare the control solution with 2.0 mL of Standard Lead 409C.
Solution (not more than 20 ppm). Mobile phase: Dissolve 17 g of sodium acetate trihydrate
(3) Arsenic <1.11>—Prepare the test solution with 1.0 g in 790 mL of water, and add 0.6 mL of acetic acid (100). If
of Cefalotin Sodium according to Method 3, and perform necessary adjust the pH to 5.9 ± 0.1 with diluted sodium
the test (not more than 2 ppm). hydroxide TS (1 in 10) or acetic acid (100). To this solution
(4) Related substances—Pipet 1 mL of the standard solu- add 150 mL of acetonitrile and 70 mL of ethanol (95).
tion obtained in the Assay, add the mobile phase to make ex- Flow rate: Adjust so that the retention time of cefalotin is
actly 100 mL, and use this solution as the standard solution. about 12 minutes.
Perform the test with exactly 10 mL each of the sample solu- System suitability—
tion obtained in the Assay and the standard solution pre- System performance: Heat the standard solution in a
pared here as directed under Liquid Chromatography <2.01> water bath of 909C for 10 minutes, and cool. Measure ex-
according to the following conditions, and determine each actly 2.5 mL of this solution, and add the mobile phase to
peak area by the automatic integration method: the area of make exactly 100 mL. When the procedure is run with 10 mL
the peaks other than cefalotin from the sample solution is of this solution under the above operating conditions, the
not larger than the peak area of cefalotin from the standard resolution between the peak of cefalotin and the peak, hav-
solution, and the total area of the peaks other than cefalotin ing the relative retention time of about 0.5 to cefalotin is not
from the sample solution is not larger than 3 times the peak less than 9, and the symmetry factor of the peak of cefalotin
area of cefalotin from the standard solution. is not more than 1.8.
Operating conditions— System repeatability: When the test is repeated 6 times
Detector, column, column temperature, mobile phase, and with 10 mL of the standard solution under the above operat-
flow rate: Proceed as directed in the operating conditions in ing conditions, the relative standard deviation of the peak
the Assay. area of cefalotin is not more than 1.0z.
retention time of cefalotin.
the standard solution, and add the mobile phase to make Cefatrizine Propylene Glycolate
exactly 10 mL. Confirm that the peak area of cefalotin ob-
セファトリジンプロピレングリコール
System performance: Heat the standard solution in a
water bath of 909C for 10 minutes, and cool. Measure ex-
actly 2.5 mL of this solution, and add the mobile phase to
make exactly 100 mL. When the procedure is run with 10 mL
of this solution under the above operating conditions, the
resolution between the peak of cefalotin and the peak, hav-
ing the relative retention time of about 0.5 to cefalotin, is C18H18N6O5S2.C3H8O2: 538.60
not less than 9, and the symmetry factor of the peak of (6R,7R)-7-[(2R)-2-Amino-2-(4-
cefalotin is not more than 1.8. hydroxyphenyl)acetylamino]-8-oxo-3-[2-(1H-1,2,3-
System repeatability: When the test is repeated 3 times triazol-4-yl)sulfanylmethyl]-5-thia-1-
with 10 mL of the standard solution under the above operat- azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
ing conditions, the relative standard deviation of the peak monopropane-1,2-diolate (1/1)
area of cefalotin is not more than 2.0z. [51627-14-6, Cefatrizine]
Cefatrizine Propylene Glycolate contains not less
606 Cefatrizine Propylene Glycolate for Syrup / Official Monographs JP XVII
than 816 mg (potency) and not more than 876 mg lene Glycolate and Cefatrizine Propylene Glycolate RS,
(potency) per mg, calculated on the anhydrous basis. equivalent to about 0.1 g (potency), dissolve each in water to
The potency of Cefatrizine Propylene Glycolate is make exactly 500 mL, and use these solutions as the sample
expressed as mass (potency) of cefatrizine solution and the standard solution, respectively. Perform the
(C18H18N6O5S2: 462.50). test with exactly 10 mL each of the sample solution and
Description Cefatrizine Propylene Glycolate occurs as a
white to yellowish white powder.
the peak areas, AT and AS, of cefatrizine in each solution.
methanol and in ethanol (95). Amount [ mg (potency)] of cefatrizine (C18H18N6O5S2)
＝ MS × AT/AS × 1000
solution of Cefatrizine Propylene Glycolate (1 in 50,000) as MS: Amount [mg (potency)] of Cefatrizine Propylene
directed under Ultraviolet-visible Spectrophotometry <2.24>, Glycolate RS taken
the spectrum of a solution of Cefatrizine Propylene Glyco-
late RS prepared in the same manner as the sample solution:
length: 270 nm).
same wavelengths.
Cefatrizine Propylene Glycolate as directed in the potassium
409C.
Mobile phase: A mixture of a solution of potassium dihy-
trum or the spectrum of Cefatrizine Propylene Glycolate RS:
drogenphosphate (17 in 12,500) and methanol (17:3).
Flow rate: Adjust so that the retention time of cefatrizine
same wave numbers.
(3) Determine the 1H spectrum of a solution of Cefatri-
zine Propylene Glycolate in a mixture of heavy water for
System performance: Dissolve about 10 mg (potency) of
nuclear magnetic resonance spectroscopy and deuterated hy-
Cefatrizine Propylene Glycolate and about 5 mg (potency) of
drochloric acid for nuclear magnetic resonance spectroscopy
Cefadroxil in 50 mL of water. When the procedure is run
(3:1) (1 in 10), using sodium 3-(trimethylsilyl)propionate-d4
for nuclear magnetic resonance spectroscopy as an internal
tions, cefadroxil and cefatrizine are eluted in this order with
reference compound, as directed under Nuclear Magnetic
the resolution between these peaks being not less than 4.
Resonance Spectroscopy <2.21>: it exhibits a double signal A
at around d 1.2 ppm, a double signal B at around d 7.0 ppm,
a double signal C at around d 7.5 ppm and a single signal D
ing conditions, the relative standard deviation of peak areas
at around d 8.3 ppm. The ratio of integrated intensity of
of cefatrizine is not more than 1.0z.
these signals, A:B:C:D, is about 3:2:2:1.
D : ＋52 – ＋589(2.5 g calculated
on the anhydrous bases, 1 mol/L hydrochloric acid TS, 50
mL, 100 mm).
Cefatrizine Propylene Glycolate for
Cefatrizine Propylene Glycolate according to Method 2, and Syrup
シロップ用セファトリジンプロピレングリコール
of Cefatrizine Propylene Glycolate according to Method 3, Cefatrizine Propylene Glycolate for Syrup is a
and perform the test (not more than 2 ppm). Use a solution preparation for syrup, which is dissolved before use.
of magnesium nitrate hexahydrate in ethanol (1 in 25). It contains not less than 90.0z and not more
(3) Related substances—Dissolve 25 mg of Cefatrizine than 105.0z of the labeled potency of Cefatrizine
Propylene Glycolate in 5 mL of water, and use this solution (C18H18N6O5S2: 462.50).
tions for Syrup, with Cefatrizine Propylene Glycolate.
as directed under Thin-layer Chromatography <2.03>. Spot Identification Powder Cefatrizine Propylene Glycolate for
5 mL each of the sample solution and standard solution on Syrup, weigh a portion of the powder, equivalent to 10 mg
a plate of silica gel for thin-layer chromatography. Develop (potency) of Cefatrizine Propylene Glycolate, and dissolve in
with a mixture of 1-butanol, water and acetic acid (100) 10 mL of water. To 2 mL of this solution add water to make
(3:1:1) to a distance of about 12 cm, and air-dry the 100 mL. Determine the absorption spectrum of this solution
plate. Spray evenly ninhydrin-citric acid-acetic acid TS on as directed under Ultraviolet-visible Spectrophotometry
the plate, and heat at 1009C for 10 minutes: the spots other <2.24>: it exhibits maxima between 225 nm and 229 nm, and
than the principal spot from the sample solution are not between 266 nm and 271 nm.
pH <2.54> Take an amount of Cefatrizine Propylene
Water <2.48> Not more than 2.0z (0.5 g, volumetric titra- Glycolate for Syrup, equivalent to 0.4 g (potency) of Cefatri-
tion, direct titration). zine Propylene Glycolate, and suspend in 10 mL of water:
the pH of this suspension is between 4.0 and 6.0.
Assay Weigh accurately an amount of Cefatrizine Propy-
JP XVII Official Monographs / Cefazolin Sodium 607
Purity Related substances—Use the sample solution ob-

tained in the Assay as the sample solution. Pipet 1 mL of the Cefazolin Sodium
sample solution, add water to make exactly 100 mL, and use
this solution as the standard solution. Perform the test with セファゾリンナトリウム
area in each solution by the automatic integration method:
the area of each peak other than cefatrizine obtained from
the sample solution is not larger than the peak area of
cefatrizine obtained from the standard solution, and the
total area of the peaks other than cefatrizine from the sam- C14H13N8NaO4S3: 476.49
ple solution is not larger than 2 times the peak area of Monosodium (6R,7R)-3-(5-methyl-1,3,4-thiadiazol-2-
cefatrizine from the standard solution. ylsulfanylmethyl)-8-oxo-7-[2-(1H-tetrazol-1-
Operating conditions— yl)acetylamino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
Detector, column, column temperature, mobile phase and carboxylate
flow rate: Proceed as directed in the operating conditions in [27164-46-1]
the Assay under Cefatrizine Propylene Glycolate.
Time span of measurement: About 2.5 times as long as the Cefazolin Sodium contains not less than 900 mg
retention time of cefatrizine, beginning after the solvent (potency) and not more than 975 mg (potency) per mg,
peak. calculated on the anhydrous basis. The potency of
System suitability— Cefazolin Sodium is expressed as mass (potency) of
System performance: Proceed as directed in the system cefazolin (C14H14N8O4S3: 454.51).
suitability in the Assay under Cefatrizine Propylene Glyco-
Description Cefazolin Sodium occurs as a white to light
late.
yellow-white, crystals or crystalline powder.
It is freely soluble in water and in formamide, slightly
soluble in methanol, and practically insoluble in ethanol
that the peak area of cefatrizine obtained from 10 mL of this
(95).
solution is equivalent to 15 to 25z of that of cefatrizine ob-
tained from 10 mL of the standard solution. Identification (1) Determine the absorption spectrum of a
System repeatability: When the test is repeated 6 times solution of Cefazolin Sodium (1 in 50,000) as directed under
with 10 mL of the standard solution under the above operat- Ultraviolet-visible Spectrophotometry <2.24>, and compare
ing conditions, the relative standard deviation of the peak the spectrum with the Reference Spectrum: both spectra
area of cefatrizine is not more than 2.0z. exhibit similar intensities of absorption at the same wave-
lengths.
Uniformity of dosage units <6.02> Cefatrizine Propylene
Glycolate for Syrup in single-dose packages meets the re-
Cefazolin Sodium as directed in the potassium bromide disk
quirement of the Mass variation test.
Assay Powder Cefatrizine Propylene Glycolate for Syrup, pare the spectrum with the Reference Spectrum: both spectra
weigh accurately a portion of the powder, equivalent to exhibit similar intensities of absorption at the same wave
about 0.1 g (potency) of Cefatrizine Propylene Glycolate, numbers.
dissolve in water to make exactly 500 mL, and use this solu- (3) Determine the 1H spectrum of a solution of Cefazolin
tion as the sample solution. Separately, weigh accurately an Sodium in heavy water for nuclear magnetic resonance spec-
amount of Cefatrizine Propylene Glycolate RS, equivalent troscopy (1 in 10), using sodium 3-trimethylsilylpropionate-
to about 20 mg (potency), dissolve in water to make exactly d4 for nuclear magnetic resonance spectroscopy as an inter-
100 mL, and use this solution as the standard solution. nal reference compound, as directed under Nuclear Magnetic
Then, proceed as directed in the Assay under Cefatrizine Resonance Spectroscopy <2.21>: it exhibits single signals, A
Propylene Glycolate. and B, at around d 2.7 ppm and at around d 9.3 ppm, re-
spectively. The ratio of integrated intensity of these signals,
Amount [mg (potency)] of cefatrizine (C18H18N6O5S2)
A:B, is about 3:1.
＝ M S × AT / AS × 5
(4) Cefazolin Sodium responds to the Qualitative Tests
MS: Amount [mg (potency)] of Cefatrizine Propylene <1.09> (1) for sodium salt.
Glycolate RS taken
D : －19 – －239(2.5 g calculated
Containers and storage Containers—Tight containers. as the anhydrous basis, water, 25 mL, 100 mm).
pH <2.54> Dissolve 1.0 g of Cefazolin Sodium in 10 mL of
water: pH of the solution is between 4.8 and 6.3.
of Cefazolin Sodium in 10 mL of water: the solution is clear
and colorless to pale yellow, and its absorbance at 400 nm
determined as directed under Ultraviolet-visible Spectropho-
tometry <2.24> is not more than 0.35. The test should be per-
formed within 10 minutes after preparing of the solution.
(2) Heavy metals <1.07>—Proceed with 2.0 g of Cefazo-
lin Sodium according to Method 2, and perform the test.
608 Cefazolin Sodium for Injection / Official Monographs JP XVII
Solution (not more than 10 ppm). Operating conditions—
(3) Arsenic <1.11>—Prepare the test solution with 2.0 g Detector: An ultraviolet absorption photometer (wave-
of Cefazolin Sodium according to Method 3, and perform length: 254 nm).
the test. When prepare the test solution, add 1.5 mL of Column: A stainless steel column 4 mm in inside diameter
hydrogen peroxide (30) after addition of 10 mL of a solution and 15 cm in length, packed with octadecylsilanized silica gel
of magnesium nitrate hexahydrate in ethanol (95) (1 in 50), for liquid chromatography (10 mm in particle diameter).
and then ignite (not more than 1 ppm). Column temperature: A constant temperature of about
(4) Related substances—Dissolve 0.10 g of Cefazolin So- 259C.
dium in 20 mL of 0.1 mol/L phosphate buffer solution (pH Mobile phase: Dissolve 2.27 g of disodium hydrogen phos-
7.0) and use this solution as the sample solution. Prepare the phate dodecahydrate and 0.47 g of citric acid monohydrate
sample solution before use. Perform the test with 5 mL of the in water to make 935 mL, and add 65 mL of acetonitrile.
sample solution as directed under Liquid Chromatography Flow rate: Adjust so that the retention time of cefazolin is
<2.01> according to the following conditions. Determine each about 8 minutes.
peak area by the automatic integration method, and calcu- System suitability—
late the amount of the peaks by the area percentage method: System performance: When the procedure is run with 5 mL
the amount of the peak, having the relative retention time of of the standard solution under the above operating condi-
about 0.2 to cefazolin and the amount of the peak other than tions, cefazolin and the internal standard are eluted in this
cefazolin and the peak mentioned above are not more than order with the resolution between these peaks being not less
1.5z, respectively. The total amount of the peaks other than than 4.
cefazolin is not more than 2.5z. For the area of the peak, System repeatability: When the test is repeated 6 times
having the relative retention time of about 0.2 to the cefazo- with 5 mL of the standard solution under the above operating
lin, multiply the relative response factor, 1.43. conditions, the relative standard deviation of the ratios of
Operating conditions— the peak area of cefazolin to that of the internal standard is
Detector, column, column temperature, mobile phase, and not more than 1.0z.
the Assay.
retention time of cefazolin, beginning after the solvent peak.
System suitability— Cefazolin Sodium for Injection
注射用セファゾリンナトリウム
Test for required detectability: Dissolve about 80 mg of
Cefazolin RS in 0.1 mol/L phosphate buffer solution (pH Cefazolin Sodium for Injection is a preparation for
7.0) to make 100 mL, and use this solution as the solution injection which is dissolved before use.
for system suitability test. Pipet 1 mL of the solution for It contains not less than 90.0z and not more
system suitability test, and add 0.1 mol/L phosphate buffer than 110.0z of the labeled potency of cefazolin
solution (pH 7.0) to make exactly 20 mL. Confirm that the (C14H14N8O4S3: 454.51).
peak area of cefazolin obtained from 5 mL of this solution is
equivalent to 3 to 7z of that obtained from 5 mL of the solu-
tions, with Cefazolin Sodium.
tion for system suitability test.
System repeatability: When the test is repeated 6 times Description Cefazolin Sodium for Injection occurs as white
with 5 mL of the solution for system suitability test under the to light yellowish white crystals or crystalline powder or
above operating conditions, the relative standard deviation masses.
of the peak areas of cefazolin is not more than 1.0z.
Water <2.48> Not more than 2.5z (1.0 g, volumetric titra- solution of Cefazolin Sodium for Injection (1 in 50,000) as
tion, direct titration. Use a mixture of formamide for water directed under Ultraviolet-visible Spectrophotometry <2.24>:
determination and methanol for water determination (2:1) it exhibits a maximum between 270 nm and 274 nm.
instead of methanol for water determination). (2) Cefazolin Sodium for Injection responds to the
Qualitative Tests <1.09> (1) for chloride.
Assay Weigh accurately an amount of Cefazolin Sodium
and Cefazolin RS, equivalent to about 20 mg (potency), dis- Osmotic pressure ratio Being specified separately when the
solve each in the internal standard solution to make exactly drug is granted approval based on the Law.
20 mL, and use these solutions as the sample solution and
the standard solution, respectively. Perform the test with 5
amount of Cefazolin Sodium for Injection, equivalent to
mL each of the sample solution and standard solution as di-
1.0 g (potency) of Cefazolin Sodium, in 10 mL of water is
4.5 to 6.5.
QS, of the peak area of cefazolin to that of the internal Purity (1) Clarity and color of solution—Conduct this
standard. procedure within 10 minutes after the preparation of the
solutions. A solution prepared by dissolving an amount of
Amount [mg (potency)] of cefazolin (C14H14N8O4S3)
Cefazolin Sodium for Injection, equivalent to 1.0 g (po-
＝ MS × QT/QS × 1000
tency) of Cefazolin Sodium, in 10 mL of water is clear, and
MS: Amount [mg (potency)] of Cefazolin RS taken the absorbance of this solution at 400 nm, determined as di-
rected under Ultraviolet-visible Spectrophotometry <2.24>, is
Internal standard solution—A solution of p-acetanisidide in
not more than 0.35.
0.1 mol/L phosphate buffer solution (pH 7.0) (11 in 20,000).
(2) Related substances—Dissolve an amount of Cefazo-
lin Sodium for Injection, equivalent to 0.10 g (potency) of
JP XVII Official Monographs / Cefazolin Sodium Hydrate 609
Cefazolin Sodium, in 20 mL of 0.1 mol/L phosphate buffer MS: Amount [mg (potency)] of Cefazolin RS taken
solution (pH 7.0) and use this solution as the sample solu-
Internal standard solution—A solution of p-acetanisidide in
tion. Prepare the sample solution before use. Perform the
0.1 mol/L phosphate buffer solution (pH 7.0) (11 in 20,000).
test with 5 mL of the sample solution as directed under Liq-
uid Chromatography <2.01> according to the following con- Containers and storage Containers—Hermetic containers.
ditions, and determine each peak area by the automatic in- Plastic containers for aqueous injections may be used.
tegration method. Calculate the amount of each peak by the
area percentage method: the amount of the peaks other than
cefazolin is not more than 1.5z. Furthermore the total Cefazolin Sodium Hydrate
amount of the peaks other than cefazolin is not more than
2.5z. For the area of the peak, having the relative retention セファゾリンナトリウム水和物
time of about 0.2 to cefazolin, multiply the relative response
factor, 1.43.
the Assay under Cefazolin Sodium.
retention time of cefazolin, beginning after the solvent peak. C14H13N8NaO4S3.5H2O: 566.57
System suitability— Monosodium (6R,7R)-3-(5-methyl-1,3,4-thiadiazol-2-
System performance: Proceed as directed in the system ylsulfanylmethyl)-8-oxo-7-[2-(1H-tetrazol-
suitability in the Assay under Cefazolin Sodium. 1-yl)acetylamino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
Test for required detectability: To 8 mL of the sample so- carboxylate pentahydrate
lution, add 0.1 mol/L phosphate buffer solution (pH 7.0) to [115850-11-8]
make 50 mL, and use this solution as the solution for system
suitability test. Pipet 1 mL of the solution for system suita- Cefazolin Sodium Hydrate contains not less than
bility test, add 0.1 mol/L phosphate buffer solution (pH 7.0) 920 mg (potency) and not more than 975 mg (potency)
to make exactly 20 mL. Confirm that the peak area of per mg, calculated on the anhydrous basis. The po-
cefazolin obtained from 5 mL of this solution is equivalent to tency of Cefazolin Sodium Hydrate is expressed as
3 to 7z of that obtained from 5 mL of the solution for sys- mass (potency) of cefazolin (C14H14N8O4S3: 454.51).
tem suitability test.
Description Cefazolin Sodium Hydrate occurs as white to
pale yellowish white crystals.
with 5 mL of the solution for system suitability test under the
slightly soluble in ethanol (95), and practically insoluble in
of the peak area of cefazolin is not more than 1.0z.
diethyl ether.
tion, direct titration). Use a mixture of formamide for water
solution of Cefazolin Sodium Hydrate (1 in 50,000) as di-
instead of methanol for water determination.
Bacterial endotoxins <4.01> Less than 0.05 EU/mg (po- both spectra exhibit similar intensities of absorption at the
tency). same wavelengths.
Cefazolin Sodium Hydrate as directed in the potassium bro-
Foreign insoluble matter <6.06> Perform the test according and compare the spectrum with the Reference Spectrum:
to Method 2: it meets the requirement. both spectra exhibit similar intensities of absorption at the
same wave numbers.
(3) Determine the 1H spectrum of a solution of Cefazolin
ment.
Sodium Hydrate in heavy water for nuclear magnetic reso-
Sterility <4.06> Perform the test according to the Mem- nance spectroscopy (1 in 10), using sodium 3-trimethylsilyl-
brane filtration method: it meets the requirement. propionate-d4 for nuclear magnetic resonance spectroscopy
as an internal reference compound, as directed under
Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits
than 10 containers of Cefazolin Sodium for Injection. Weigh
single signals, A and B, at around d 2.7 ppm and at around d
accurately an amount of the contents, equivalent to about 50
9.3 ppm. The ratio of integrated intensity of each signal,
mg (potency) of Cefazolin Sodium, dissolve in the internal
A:B, is about 3:1.
standard solution to make exactly 50 mL, and use this solu-
(4) Cefazolin Sodium Hydrate responds to the Qualita-
tive Tests <1.09> (1) for sodium salt.
amount of Cefazolin RS, equivalent to about 50 mg (po-
tency), dissolve in the internal standard solution to make Absorbance <2.24> E 11zcm (272 nm): 272 – 292 (80 mg calcu-
exactly 50 mL, and use this solution as the standard solution. lated on the anhydrous basis, water, 5000 mL).
Then, proceed as directed in the Assay under Cefazolin
D : －20 – －259(2.5 g calculated
Sodium.
Amount [mg (potency)] of cefazolin (C14H14N8O4S3)
pH <2.54> Dissolve 1.0 g of Cefazolin Sodium Hydrate in
＝ M S × Q T / QS
10 mL of water: the pH of the solution is between 4.8 and
610 Cefbuperazone Sodium / Official Monographs JP XVII
6.3. uid Chromatography <2.01> according to the following con-
of cefazolin to that of the internal standard.
of Cefazolin Sodium Hydrate in 10 mL of water: the solu-
tion is clear, and when determine the absorbance at 400 nm Amount [ mg (potency)] of cefazolin (C14H14N8O4S3)
of this solution as directed under Ultraviolet-visible Spectro- ＝ MS × QT/QS × 1000
photometry <2.24>, it is not more than 0.15.
MS: Amount [mg (potency)] of Cefazolin RS taken
(2) Heavy metals <1.07>—Proceed with 2.0 g of Cefazo-
lin Sodium Hydrate according to Method 2, and perform the Internal standard solution—A solution of p-acetanisidide in
test. Prepare the control solution with 2.0 mL of Standard 0.1 mol/L phosphate buffer solution (pH 7.0) (11 in 20,000).
Lead Solution (not more than 10 ppm). Operating conditions—
(3) Related substances—Dissolve 0.10 of Cefazolin So- Detector: An ultraviolet absorption photometer (wave-
dium Hydrate in 20 mL of 0.1 mol/L phosphate buffer solu- length: 254 nm).
tion (pH 7.0) and use this solution as the sample solution. Column: A stainless steel column 4 mm in inside diameter
Perform the test with 5 mL of the sample solution as directed and 15 cm in length, packed with octadecylsilanized silica gel
under Liquid Chromatography <2.01> according to the fol- for liquid chromatography (10 mm in particle diameter).
lowing conditions. Determine each peak area by the auto- Column temperature: A constant temperature of about
matic integration method, and calculate the amount of them 259C.
by the area percentage method: the amount of the peak hav- Mobile phase: Dissolve 2.27 g of disodium hydrogen phos-
ing the relative retention time of about 0.2 to cefazolin is not phate dodecahydrate and 0.47 g of citric acid monohydrate
more than 1.0z, the amount of the peak other than cefazo- in water to make 935 mL. To this solution, add 65 mL of
lin and the peak mentioned above is not more than 0.5z, acetonitrile.
and the total amount of the peaks other than cefazolin is not Flow rate: Adjust so that the retention time of cefazolin is
more than 2.0z. For the area of the peak, having the rela- about 8 minutes.
tive retention time of about 0.2 to cefazolin, multiply the System suitability—
relative response factor 1.43. System performance: When the procedure is run with 5 mL
Operating conditions— of the standard solution under the above operating condi-
Detector, column, column temperature, mobile phase, and tions, cefazolin and the internal standard are eluted in this
flow rate: Proceed as directed in the operating conditions in order with the resolution between these peaks being not less
the Assay. than 4.
Time span of measurement: About 3 times as long as the System repeatability: When the test is repeated 6 times
retention time of cefazolin, beginning after the solvent peak. with 5 mL of the standard solution under the above operating
System suitability— conditions, the relative standard deviation of the peak area
Test for required detectability: To 1 mL of the sample so- of cefazolin is not more than 1.0z.
lution add 0.1 mol/L phosphate buffer solution (pH 7.0) to
make 100 mL, and use this solution as the solution for sys-
tem suitability test. Pipet 1 mL of the solution for system
suitability test, add 0.1 mol/L phosphate buffer solution
(pH 7.0) to make exactly 10 mL. Confirm that the peak area
of cefazolin obtained with 5 mL of this solution is equivalent Cefbuperazone Sodium
to 7 to 13z of that obtained with 5 mL of the solution for
セフブペラゾンナトリウム
system suitability test.
System performance: Dissolve 20 mg of Cefazolin Sodium
Hydrate in 20 mL of a solution of p-acetanisidide in 0.1
mol/L phosphate buffer solution (pH 7.0) (11 in 20,000).
When the procedure is run with 5 mL of this solution
under the above operating conditions, cefazolin and p-
acetanisidide are eluted in this order with the resolution
between these peaks being not less than 4.
C22H28N9NaO9S2: 649.63
Monosodium (6R,7S )-7-{(2R,3S )-2-[(4-ethyl-2,3-
dioxopiperazine-1-carbonyl)amino]-3-
of the peak area of cefazolin is not more than 2.0z.
hydroxybutanoylamino}-7-methoxy-3-(1-methyl-1H-
Water <2.48> Not less than 13.7z and not more than tetrazol-5-ylsulfanylmethyl)-8-oxo-5-thia-1-
16.0z (0.1 g, volumetric titration, direct titration. Use a azabicyclo[4.2.0]oct-2-ene-2-carboxylate
mixture of formamide for water determination and metha- [76648-01-6]
nol for water determination (2:1) instead of methanol for
water determination). Cefbuperazone Sodium contains not less than 870
mg (potency) per mg, calculated on the anhydrous
basis. The potency of Cefbuperazone Sodium is
tency).
expressed as mass (potency) of cefbuperazone
Assay Weigh accurately an amount of Cefazolin Sodium (C22H29N9O9S2: 627.65).
Hydrate and Cefazolin RS, equivalent to about 20 mg (po-
Description Cefbuperazone Sodium occurs as white to light
tency), dissolve in exactly 20 mL of the internal standard so-
yellowish white, powder or masses.
lution, and use these solutions as the sample solution and
It is very soluble in water, freely soluble in methanol and
in pyridine, sparingly soluble in ethanol (95), and very
JP XVII Official Monographs / Cefbuperazone Sodium 611
slightly soluble in acetonitrile. exactly 10 mL. Confirm that the peak area of cefbuperazone
solution of Cefbuperazone Sodium (1 in 50,000) as directed
factor of the peak of cefbuperazone are not less than 5000
wavelengths.
(2) Dissolve 0.1 g of Cefbuperazone Sodium in 0.5 mL
of deuterated pyridine for nuclear magnetic resonance spec-
troscopy and 1 drop of heavy water for nuclear magnetic
resonance spectroscopy, and determine the 1H spectrum of
area of cefbuperazone is not more than 2.0z.
this solution as directed under Nuclear Magnetic Resonance
Spectroscopy <2.21>, using tetramethylsilane for nuclear Water <2.48> Not more than 1.0z (3 g, volumetric titra-
magnetic resonance spectroscopy as an internal reference tion, direct titration).
compound: it exhibits a triplet signal A at around d 1.1 ppm,
Assay Weigh accurately an amount of Cefbuperazone So-
and two doublet signals, B and C, at around d 1.6 ppm and
dium and Cefbuperazone RS, equivalent to about 0.1 g (po-
at around d 5.1 ppm, respectively. The ratio of the inte-
tency), and dissolve each in the mobile phase to make exactly
grated intensity of each signal, A:B:C, is about 3:3:1.
100 mL. Measure exactly 10 mL each of these solutions, add
(3) Cefbuperazone Sodium responds to the Qualitative
exactly 10 mL of the internal standard solution and the
mobile phase to make 50 mL, and use these solutions as the
D : ＋48 – ＋569(0.4 g calculated sample solution and standard solution. Perform the test with
on the anhydrous basis, water, 20 mL, 100 mm). 10 mL each of the sample solution and standard solution as
pH <2.54> Dissolve 1.0 g of Cefbuperazone Sodium in 4
QS, of the peak area of cefbuperazone to that of the internal
Purity (1) Clarity and color of solution—Dissolve 1.0 g standard.
of Cefbuperazone Sodium in 4 mL of water: the solution is
Amount [ mg (potency)] of cefbuperazone (C22H29N9O9S2)
clear and light yellow.
＝ MS × QT/QS × 1000
(2) Heavy metals <1.07>—Proceed with 2.0 g of Cef-
buperazone Sodium according to Method 4, and perform the MS: Amount [mg (potency)] of Cefbuperazone RS taken
Internal standard solution—A solution of acetanilide in the
of Cefbuperazone Sodium according to Method 4, and per-
length: 254 nm).
(4) Related substances—Dissolve 0.10 g of Cefbupera-
zone Sodium in 100 mL of the mobile phase, and use this so-
259C.
Mobile phase: Dissolve 2.0 g of tetra-n-propylammonium
bromide in 1000 mL of a mixture of water, acetonitrile
and acetic acid-sodium acetate buffer solution (pH 5.0)
(83:13:4).
the percentages of each peak area of related substances from
Flow rate: Adjust so that the retention time of cefbupera-
the sample solution against 50 times of the peak area of
zone is about 16 minutes.
cefbuperazone from the standard solution; the amount of
related substance I having the relative retention time of
about 0.2 to cefbuperazone is not more than 2.0z, the
amount of related substance II having the relative retention
ditions, the internal standard and cefbuperazone are eluted
time of about 0.6 to cefbuperazone is not more than 4.5z
and the amount of related substance III having the relative
not less than 3.
retention time of about 1.6 to cefbuperazone is not more
than 1.0z, and the total amount of these related substances
is not more than 6.0z. For the peak areas of the related sub-
stances I and III, multiply their relative response factors,
of the peak area of cefbuperazone to that of the internal
0.72 and 0.69, respectively.
Detector, column, column temperature, mobile phase, and Containers and storage Containers—Hermetic containers.
flow rate: Proceed as directed in the operating conditions in Storage—In a cold place.
the Assay.
retention time of cefbuperazone.
612 Cefcapene Pivoxil Hydrochloride Hydrate / Official Monographs JP XVII
with 2.0 mL of Standard Lead Solution (not more than 10
Cefcapene Pivoxil Hydrochloride ppm).
(2) Related substance I—Dissolve an amount of Cefca-
Hydrate pene Pivoxil Hydrochloride Hydrate, equivalent to about 10
mg (potency), in 2 mL of methanol, add a mixture of water
セフカペンピボキシル塩酸塩水和物
and methanol (1:1) to make 50 mL, and use this solution as
the sample solution. Perform the test with 30 mL of the sam-
ple solution as directed under Liquid Chromatography
each peak area by the automatic integration method. If nec-
essary, compensate the base-line by performing in the same
manner as the test with 30 mL of a mixture of water and
methanol (1:1). Measure the amount of the peak other than
cefcapene pivoxil by the area percentage method: the
C23H29N5O8S2.HCl.H2O: 622.11
amounts of the peaks, having the relative retention times of
2,2-Dimethylpropanoyloxymethyl (6R,7R)-7-[(2Z )-2-(2-
about 1.5 and about 1.7 to cefcapene pivoxil, are not more
aminothiazol-4-yl)pent-2-enoylamino]-3-
than 0.2z, respectively. The amount of the peaks other than
carbamoyloxymethyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-
the peaks mentioned above is not more than 0.1z, and the
ene-2-caboxylate monohydrochloride monohydrate
total of them is not more than 1.5z.
[147816-24-8]
Cefcapene Pivoxil Hydrochloride Hydrate contains
length: 265 nm).
not less than 722 mg (potency) and not more than 764
mg (potency) per mg, calculated on the anhydrous
basis. The potency of Cefcapene Pivoxil Hydrochlo-
ride Hydrate is expressed as mass (potency) of cefca-
pene (C17H19N5O6S2: 453.49).
209C.
Description Cefcapene Pivoxil Hydrochloride Hydrate oc- Mobile phase A: Dissolve 5.99 g of potassium dihydrogen
curs as a white to pale yellowish white, crystalline powder or phosphate in water to make 1100 mL. To this solution add a
mass. It has slightly a characteristic odor. solution prepared by dissolving 1.89 g of tetra-n-pentylam-
It is freely soluble in N, N-dimethylformamide and in monium bromide in methanol to make 1000 mL.
methanol, soluble in ethanol (99.5), slightly soluble in water, Mobile phase B: A mixture of methanol and water (22:3).
and practically insoluble in diethyl ether. Flowing of mobile phase: Control the gradient by mixing
solution of Cefcapene Pivoxil Hydrochloride Hydrate in
methanol (1 in 50,000) as directed under Ultraviolet-visible Time after injection Mobile phase A Mobile phase B
Spectrophotometry <2.24>, and compare the spectrum with of sample (min) (volz) (volz)
the Reference Spectrum or the spectrum of a solution of Cef-
0 – 20 98 2
capene Pivoxil Hydrochloride RS prepared in the same man-
20 – 40 98 → 50 2 → 50
ner as the sample solution: both spectra exhibit similar inten-
40 – 50 50 50
sities of absorption at the same wavelengths.
(2) Determine the infrared absorption spectra of Cefca-
pene Pivoxil Hydrochloride Hydrate and Cefcapene Pivoxil Flow rate: 0.8 mL per minute.
Hydrochloride RS as directed in the paste method under In- Time span of measurement: About 2.5 times as long as the
frared Spectrophotometry <2.25>, and compare these spec- retention time of cefcapene pivoxil.
tra: both spectra exhibit similar intensities of absorption at System suitability—
the same wave numbers. Test for required detectability: To exactly 1 mL of the
(3) Determine the 1H spectrum of a solution of Cefca- sample solution add a mixture of water and methanol (1:1)
pene Pivoxil Hydrochloride Hydrate in deuterated methanol to make 100 mL, and use this solution as the solution for
for nuclear magnetic resonance spectroscopy (1 in 50) as system suitability test. Pipet 1 mL of the solution for system
directed under Nuclear Magnetic Resonance Spectroscopy suitability test, and add the mixture of water and methanol
<2.21>, using tetramethylsilane for nuclear magnetic reso- (1:1) to make exactly 10 mL. Confirm that the peak area of
nance spectroscopy as an internal reference compound: it cefcapene pivoxil obtained from 30 mL of this solution is
exhibits a triplet signal A at around d 6.3 ppm, and a single equivalent to 7 to 13z of that of cefcapene pivoxil obtained
signal B at around d 6.7 ppm, and the ratio of integrated from 30 mL of the solution for system suitability test.
intensity of each signal, A:B, is about 1:1. System performance: Dissolve 10 mg of Cefcapene Pivoxil
(4) Dissolve 10 mg of Cefcapene Pivoxil Hydrochloride Hydrochloride Hydrate and 10 mg of propyl parahydroxy-
Hydrate in 2 mL of a mixture of water and methanol (1:1), benzoate in 25 mL of methanol, and add water to make 50
and add 1 drop of silver nitrate TS: a white precipitate is mL. To 5 mL of this solution add the mixture of water and
formed. methanol (1:1) to make 50 mL. When the procedure is run
D : ＋51 – ＋549(0.1 g calculated tions, cefcapene pivoxil and propyl parahydroxybenzoate are
on the anhydrous basis, methanol, 10 mL, 100 mm). eluted in this order with the resolution between these peaks
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of being not less than 7.
Cefcapene Pivoxil Hydrochloride Hydrate according to System repeatability: When the test is repeated 3 times
Method 4, and perform the test. Prepare the control solution with 30 mL of the solution for system suitability test under
JP XVII Official Monographs / Cefcapene Pivoxil Hydrochloride Fine Granules 613
the above operating conditions, the relative standard devia- Amount [ mg (potency)] of cefcapene (C17H19N5O6S2)
tion of the peak area of cefcapene pivoxil is not more than ＝ MS × QT/QS × 1000
4.0z.
MS: Amount [mg (potency)] of Cefcapene Pivoxil Hydro-
(3) Related substance II—Dissolve an amount of Cefca-
chloride RS taken
pene Pivoxil Hydrochloride Hydrate, equivalent to about 2
mg (potency), in N, N-dimethylformamide for liquid chro- Internal standard solution—A solution of p-benzylphenol in
matography to make 20 mL, and use this solution as the a mixture of water and methanol (1:1) (7 in 4000).
sample solution. Perform the test with 20 mL of the sample Operating conditions—
solution as directed under Liquid Chromatography <2.01> Detector: An ultraviolet absorption photometer (wave-
according to the following conditions, and determine each length: 265 nm).
peak area by the automatic integration method: the total Column: A stainless steel column 3.0 mm in inside diame-
area of the peaks which appear earlier than cefcapene pivoxil ter and 7.5 cm in length, packed with octadecylsilanized
is not more than 1.7z of the total area of the peaks other silica gel for liquid chromatography (3 mm in particle diame-
than the solvent. ter).
Operating conditions— Column temperature: A constant temperature of about
Detector: An ultraviolet absorption photometer (wave- 409C.
length: 280 nm). Mobile phase: Dissolve 1.56 g of sodium dihydrogenphos-
Column: A stainless steel column 7.8 mm in inside diame- phate dihydrate and 1.22 g of sodium 1-decanesulfonate in
ter and 30 cm in length, packed with styrene-divinylbenzene water to make 1000 mL. To 700 mL of this solution add 300
copolymer for liquid chromatography. mL of acetonitrile and 100 mL of methanol.
Column temperature: A constant temperature of about Flow rate: Adjust so that the retention time of cefcapene
259 C. pivoxil is about 5 minutes.
Mobile phase: A solution of lithium bromide in N, N- System suitability—
dimethylformamide for liquid chromatography (13 in 5000). System performance: Dissolve 0.2 g of Cefcapene Pivoxil
Flow rate: Adjust so that the retention time of cefcapene Hydrochloride Hydrate in 10 mL of methanol, and warm in
pivoxil is about 22 minutes. a water bath at 609 C for 20 minutes. After cooling, pipet 1
Time span of measurement: About 1.8 times as long as the mL of this solution, and add exactly 10 mL of the internal
retention time of cefcapene pivoxil. standard solution and the mixture of water and methanol
System suitability— (1:1) to make 50 mL. When the procedure is run with 10 mL
Test for required detectability: To exactly 1 mL of the of this solution under the above operating conditions, cefca-
sample solution add N, N-dimethylformamide for liquid pene pivoxil, trans-cefcapene pivoxil and the internal stand-
chromatography to make 100 mL, and use this solution as ard are eluted in this order, the relative retention time of
the solution for system suitability test. Pipet 3 mL of the so- trans-cefcapene pivoxil and the internal standard to that of
lution for system suitability test, and add N, N-dimethylfor- cefcapene pivoxil are about 1.7 and about 2.0, respectively,
mamide for liquid chromatography to make exactly 10 mL. and the resolution between the peaks of trans-cefcapene
Conform that the peak area of cefcapene pivoxil obtained pivoxil and the internal standard is not less than 1.5.
from 20 mL of this solution is equivalent to 20 to 40z of that System repeatability: When the test is repeated 5 times
of cefcapene pivoxil obtained from 20 mL of the solution for with 10 mL of the standard solution under the above operat-
system suitability test. ing conditions, the relative standard deviation of the ratio of
System performance: When the procedure is run with 20 the peak area of cefcapene pivoxil to that of the internal
mL of the sample solution under the above operating condi- standard is not more than 1.0z.
tions, the number of theoretical plates of the peak of cefca-
pene pivoxil is not less than 12,000.
Storage—Light-resistant, at a temperature not exceeding
C.
59
tion of the peak areas of cefcapene pivoxil is not more than
4.0z. Cefcapene Pivoxil Hydrochloride
Water <2.48> Not less than 2.8z and not more than 3.7z Fine Granules
(0.5 g, volumetric titration, back titration).
セフカペンピボキシル塩酸塩細粒
Assay Weigh accurately an amount of Cefcapene Pivoxil
Hydrochloride Hydrate and Cefcapene Pivoxil Hydrochlo-
Cefcapene Pivoxil Hydrochloride Fine Granules
ride RS, equivalent to about 20 mg (potency), and dissolve
contain not less than 90.0z and not more than
each in a mixture of water and methanol (1:1) to make
110.0z of the labeled potency of cefcapene
exactly 50 mL. Pipet 10 mL each of these solutions, add
(C17H19N5O6S2: 453.49).
exactly 10 mL of the internal standard solution and the
mixture of water and methanol (1:1) to them to make 50 mL, Method of preparation Prepare as directed under Gran-
and use these solutions as the sample solution and the stand- ules, with Cefcapene Pivoxil Hydrochloride Hydrate.
ard solution, respectively. Perform the test with 10 mL each
Identification Powder Cefcapene Pivoxil Hydrochloride
Fine Granules. To a portion of the powder, equivalent to 10
mg (potency) of Cefcapene Pivoxil Hydrochloride Hydrate,
add 40 mL of methanol, shake vigorously, and add metha-
the peak area of cefcapene pivoxil to that of the internal
nol to make 50 mL. To 4 mL of this solution add methanol
standard.
to make 50 mL, and filter through a membrane filter with a
pore size of 0.45 mm. Determine the absorption spectrum of
614 Cefcapene Pivoxil Hydrochloride Tablets / Official Monographs JP XVII
the filtrate as directed under Ultraviolet-visible Spectropho- add 100 mL of the mixture of water and methanol (1:1),
tometry <2.24>: it exhibits a maximum between 264 nm and shake vigorously for 10 minutes, add the mixture of water
268 nm. and methanol (1:1) to make exactly 200 mL, and centrifuge
at 3000 rpm for 5 minutes. Filter the supernatant liquid
Purity (1) Related substances I—Powder Cefcapene
through a membrane filter with a pore size of 0.45 mm, dis-
Pivoxil Hydrochloride Fine Granules. To a portion of the
powder, equivalent to 5 mg (potency) of Cefcapene Pivoxil
Hydrochloride Hydrate, add 1 mL of methanol, and shake.
lution and the mixture of water and methanol (1:1) to make
Add 25 mL of a mixture of water and methanol (1:1), shake
vigorously for 5 minutes, and filter through a membrane
rately, weigh accurately about 20 mg (potency) of Cefcapene
filter with a pore size of 0.45 mm. Discard the first 3 mL of
Pivoxil Hydrochloride RS, and dissolve in the mixture of
the filtrate, and use the subsequent filtrate as the sample
water and methanol (1:1) to make exactly 50 mL. Pipet 10
solution. Perform the test with 30 mL of the sample solution
mL of this solution, add exactly 10 mL of the internal stand-
ard solution and the mixture of water and methanol (1:1) to
to the following conditions. Determine each peak area by the
automatic integration method. If necessary, compensate the
Proceed as directed in the Assay under Cefcapene Pivoxil
base-line by performing in the same manner as the test with
Hydrochloride Hydrate.
30 mL of a mixture of water and mothod (1:1). Calculate the
amount of the peaks other than the peak of cefcapene Amount [mg (potency)] of cefcapene (C17H19N5O6S2)
pivoxil by the area percentage method: the amount of the ＝ MS × QT/QS × 10
substance, having the relative retention time of about 1.3 to
cefcapene pivoxil, is not more than 0.4z, the amount of the
chloride RS taken
trans-isomer of cefcapene pivoxil, having the relative reten-
tion time of about 1.5, is not more than 1.1z, the amount Internal standard solution—A solution of p-benzylphenol in
of the substance other than that mentioned above is not the mixture of water and methanol (1:1) (7 in 4000).
more than 0.3z, and the total amount of these substances is
not more than 2.8z.
Purity (2) under Cefcapene Pivoxil Hydrochloride Hydrate.
System suitability— Cefcapene Pivoxil Hydrochloride
Proceed as directed in the system suitability in the Purity Tablets
(2) under Cefcapene Pivoxil Hydrochloride Hydrate.
(2) Related substances II—Powder Cefcapene Pivoxil セフカペンピボキシル塩酸塩錠
Hydrochloride Fine Granules. To a portion of the powder,
equivalent to 2 mg (potency) of Cefcapene Pivoxil Hydro-
Cefcapene Pivoxil Hydrochloride Tablets contain
chloride Hydrate, add 20 mL of N, N-dimethylformamide
not less than 90.0z and not more than 105.0z of the
for liquid chromatography, shake vigorously for 10 minutes,
labeled potency of cefcapene (C17H19N5O6S2: 453.49).
and filter through a membrane filter with a pore size of 0.45
mm. Discard the first 3 mL of the filtrate, and use the subse- Method of preparation Prepare as directed under Tablets,
quent filtrate as the sample solution. Perform the test with with Cefcapene Pivoxil Hydrochloride Hydrate.
20 mL of the sample solution as directed under Liquid Chro-
Identification To an amount of powdered Cefcapene
Pivoxil Hydrochloride Tablets, equivalent to about 10 mg
and determine each peak area by the automatic integration
(potency) of Cefcapene Pivoxil Hydrochloride Hydrate, add
method: the total area of the peaks eluted before that of cef-
40 mL of methanol, shake vigorously, and add methanol to
capene pivoxil is not more than 4.0z of the total area of all
make 50 mL. To 4 mL of this solution add methanol to
peaks other than the solvent peak.
make 50 mL, filter through a membrane filter with pore size
of 0.45 mm, and use the filtrate as the sample solution. De-
Proceed as directed in the operating conitions in the Purity
termine the absorption spectrum of the sample solution as
directed under Ultraviolet-visible Spectrophotometry <2.24>:
it exhibits a maximum between 263 nm and 267 nm.
(3) under Cefcapene Pivoxil Hydrochloride Hydrate. Purity (1) Related substances I—To an amount of pow-
dered Cefcapene Pivoxil Hydrochloride Tables, equivalent
to about 5 mg (potency) of Cefcapene Pivoxil Hydrochloride
tion, back titration). Perform the test without pulverizing
Hydrate, add 1 mL of methanol, and shake. Add 25 mL of a
the sample, and handling the sample under a relative hu-
mixture of water and methanol (1:1), shake vigorously for 5
midity of less than 30z.
minutes, and filter through a membrane filter with pore size
Uniformity of dosage units <6.02> The granules in single- of 0.45 mm. Discard the first 3 mL of the filtrate, and use the
dose packages meet the requirement of the Mass variation subsequent filtrate as the sample solution. Perform the test
test. with 30 mL of the sample solution as directed under Liquid
tegration method. If necessary, proceed with 30 mL of the
Assay Weigh accurately an amount of Cefcapene Pivoxil mixture of water and methanol (1:1) in the same manner as
Hydrochloride Fine Granules, equivalent to about 0.2 g (po- the sample solution to compensate the base line. Calculate
tency) of and Cefcapene Pivoxil Hydrochloride Hydrate, the amounts of the peaks other than cefcapene pivoxil by the
JP XVII Official Monographs / Cefdinir 615
area percentage method: the amount of the peak, having the Tablets, equivalent to about 0.6 g (potency) of Cefcapene
relative retention time of about 1.3 to cefcapene pivoxil, is Pivoxil Hydrochloride Hydrate, add 20 mL of water, and
not more than 0.4z, the amount of the peak of cefcapene shake for 5 minutes to disintegrate. Add 80 mL of methanol,
pivoxil trans-isomer, having the relative retention time of shake vigorously for 5 minutes, add a mixture of methanol
about 1.5, is not more than 0.5z, the amount of the peaks and water (4:1) to make exactly 200 mL, and centrifuge at
other than the peaks mentioned above are not more than 3000 rpm for 5 minutes. Filter the supernatant liquid
0.3z, respectively, and the total amount of these peaks is through a membrane filter with pore size of 0.45 mm, and
not more than 2.0z. discard the first 1 mL of the filtrate. Pipet 2 mL of the sub-
Operating conditions— sequent filtrate, add exactly 15 mL of the internal standard
Proceed as directed in the operating conditions in the solution, add the mixture of water and methanol (1:1) to
Purity (2) under Cefcapene Pivoxil Hydrochloride Hydrate. make 75 mL, and use this solution as the sample solution.
System suitability— Separately, weigh accurately an amount of Cefcapene
Proceed as directed in the system suitability in the Purity Pivoxil Hydrochloride RS, equivalent to about 20 mg (po-
(2) under Cefcapene Pivoxil Hydrochloride Hydrate. tency), and dissolve in the mixture of water and methanol
(2) Related substances II—To an amount of powdered (1:1) to make exactly 50 mL. Pipet 10 mL of this solution,
Cefcapene Pivoxil Hydrochloride Tablets, equivalent to 2 add exactly 10 mL of the internal standard solution, add the
mg (potency) of Cefcapene Pivoxil Hydrochloride Hydrate, mixture of water and methanol (1:1) to make 50 mL, and use
add 20 mL of N, N-dimethylformamide for liquid chroma- this solution as the standard solution. Proceed as directed in
tography, shake vigorously for 10 minutes, and filter the Assay under Cefcapene Pivoxil Hydrochloride Hydrate.
through a membrane filter with pore size of 0.45 mm. Dis-
Amount [mg (potency)] of cefcapene (C17H19N5O6S2)
card the first 3 mL of the filtrate, and use the subsequent fil-
＝ MS × QT/QS × 30
trate as the sample solution. Perform the test with 20 mL of
the sample solution as directed under Liquid Chromatogra- MS: Amount [mg (potency)] of Cefcapene Pivoxil Hydro-
phy <2.01> according to the following conditions, and deter- chloride RS taken
mine each peak area by the automatic integration method:
Internal standard solution—A solution of p-benzylphenol in
the total area of the peaks which are eluted before cefcapene
the mixture of water and methanol (1:1) (7 in 4000).
pivoxil is not more than 3.3z of the total area of the peaks
other than the solvent peak. Containers and storage Containers—Tight containers.
Purity (3) under Cefcapene Pivoxil Hydrochloride Hydrate. Cefdinir
Proceed as directed in the system suitability in the Purity セフジニル
tion, back titration). Powdering of the sample tablets and
handling of the powder are performed under the relative
humidity of not exceeding 30z.
C14H13N5O5S2: 395.41
(6R,7R)-7-[(Z )-2-(2-Aminothiazol-4-yl)-
2-(hydroxyimino)acetylamino]-8-oxo-3-vinyl-5-thia-1-
To 1 tablet of Cefcapene Pivoxil Hydrochloride Tablets
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
add 5 mL of water, and shake vigorously for 5 minutes to
[91832-40-5]
disintegrate. Add 20 mL of methanol, shake vigorously for 5
minutes, add a mixture of methanol and water (4:1) to make
Cefdinir contains not less than 930 mg (potency) and
exactly 50 mL, and centrifuge at 3000 rpm for 5 minutes.
not more than 1020 mg (potency) per mg. The potency
Filter the supernatant liquid through a membrane filter with
of Cefdinir is expressed as mass (potency) of cefdinir
pore size of 0.45 mm, and discard the first 1 mL of the fil-
(C14H13N5O5S2).
trate. Pipet V mL of the subsequent filtrate, equivalent to
about 6 mg (potency) of Cefcapene Pivoxil Hydrochloride Description Cefdinir occurs as a white to light yellow crys-
Hydrate, add exactly 15 mL of the internal standard solu- talline powder.
tion, add a mixture of water and methanol (1:1) to make 75 It is practically insoluble in water, in ethanol (95) and in
mL, and use this solution as the sample solution. Then, diethyl ether.
proceed as directed in the Assay. It dissolves in 0.1 mol/L phosphate buffer solution (pH
7.0).
Amount [mg (potency)] of cefcapene (C17H19N5O6S2)
＝ MS × QT/QS × 15/V Identification (1) Determine the absorption spectra of so-
lutions of Cefdinir and Cefdinir RS in 0.1 mol/L phosphate
buffer solution (pH 7.0) (1 in 100,000) as directed under Ul-
chloride RS taken
traviolet-visible Spectrophotometry <2.24>, and compare
Internal standard solution—A solution of p-benzylphenol in these spectra: both spectra exhibit similar intensities of ab-
a mixture of water and methanol (1:1) (7 in 4000). sorption at the same wavelengths.
(2) Determine the infrared absorption spectra of Cef-
dinir and Cefdinir RS as directed in the paste method under
Infrared Spectrophotometry <2.25>, and compare these spec-
Assay To an amount of Cefcapene Pivoxil Hydrochloride tra: both spectra exhibit similar intensities of absorption at
616 Cefdinir / Official Monographs JP XVII
the same wave numbers. tion, beginning after the solvent peak.
(3) Determine the 1H spectrum of a solution of Cefdinir System suitability—
in a mixture of deuterated dimethyl sulfoxide for nuclear Test for required detectability: To 1 mL of the sample so-
magnetic resonance spectroscopy and heavy water for lution add tetramethylammonium hydroxide TS (pH 5.5) to
nuclear magnetic resonance spectroscopy (4:1) (1 in 10), make 100 mL, and use this solution as the solution for sys-
using tetramethylsilane for nuclear magnetic resonance spec- tem suitability test. Pipet 1 mL of the solution for system
troscopy as an internal reference compound, as directed suitability test, add tetramethylammonium hydroxide TS
under Nuclear Magnetic Resonance Spectroscopy <2.21>: it (pH 5.5) to make exactly 10 mL. Confirm that the peak area
exhibits multiple signals, A at around d 5.0 – 6.1 ppm and B of cefdinir obtained from 10 mL of this solution is equivalent
at around d 6.4 – 7.5 ppm. The ratio of integrated intensity to 7 to 13z of that obtained from 10 mL of the solution for
of each signal, A:B is about 2:1. system suitability test.
System performance: Dissolve 30 mg of Cefdinir RS and 2
D : －58 – －669 (0.25 g, 0.1
mg of cefdinir lactam ring-cleavage lactones in 3 mL of 0.1
mol/L phosphate buffer solution (pH 7.0), 25 mL, 100 mm).
mol/L phosphate buffer solution (pH 7.0), add tetramethy-
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of lammonium hydroxide TS (pH 5.5) to make 20 mL. When
Cefdinir according to Method 2, and perform the test. Pre- the procedure is run with 10 mL of this solution under the
pare the control solution with 2.0 mL of Standard Lead So- above operating conditions, peak 1 and peak 2 of cefdinir
lution (not more than 10 ppm). lactam ring-cleavage lactones separated into 4 peaks,
(2) Related substances—Dissolve about 0.1 g of Cefdinir cefdinir, peak 3 and peak 4 of remaining cefdinir lactam
in 10 mL of 0.1 mol/L phosphate buffer solution (pH 7.0). ring-cleavage lactones are eluted in this order. Relative reten-
To 3 mL of this solution add tetramethylammonium hydrox- tion time of peak 3 of cefdinir lactam ring-cleavage lactone
ide TS (pH 5.5) to make 20 mL, and use this solution as the to cefdinir is about 1.11. The number of theoretical plates
sample solution. Perform the test with 10 mL of the sample and the symmetry factor of the peak of cefdinir are not less
solution as directed under Liquid Chromatography <2.01> than 7000 and not more than 3.0, respectively.
according to the following conditions. Determine each peak System repeatability: When the test is repeated 3 times
area by the automatic integration method, and calculate the with 10 mL of the solution for system suitability test under
amounts of their peaks by the area percentage method: the the above operating conditions, the relative standard devia-
amount of the peaks, having the relative retention time of tion of the peak areas of cefdinir is not more than 2.0z.
about 0.7, about 1.2 and about 1.5 to cefdinir, are not more
than 0.7z, not more than 0.3z and not more than 0.8z,
tion, direct titration. Use a mixture of formamide for water
respectively, the total amount of the peaks, having the rela-
tive retention time of about 0.85, about 0.93, about 1.11 and
instead of methanol for water determination).
about 1.14 to cefdinir, is not more than 0.4z, and the
amount of the peak other than cefdinir and the peaks men- Assay Weigh accurately an amount of Cefdinir and Cef-
tioned above is not more than 0.2z. And the total amount dinir RS equivalent to about 20 mg (potency), dissolve each
of the peaks other than cefdinir is not more than 3.0z. in 0.1 mol/L phosphate buffer solution (pH 7.0) to make ex-
Operating conditions— actly 100 mL, and use these solutions as the sample solution
Detector: An ultraviolet absorption photometer (wave- and standard solution. Perform the test with exactly 5 mL of
length: 254 nm). the sample solution and standard solution as directed under
Column: A stainless steel column 4.6 mm in inside diame- Liquid Chromatography <2.01> according to the following
ter and 15 cm in length, packed with octadecylsilanized silica conditions, and determine the peak areas, AT and AS, of
gel for liquid chromatography (5 mm in particle diameter). cefdinir in each solution.
Amount [ mg (potency)] of cefdinir (C14H13N5O5S2)
409 C.
＝ MS × AT/AS × 1000
Mobile phase A: To 1000 mL of tetramethylammonium
hydroxide TS (pH 5.5) add 0.4 mL of 0.1 mol/L disodium MS: Amount [mg (potency)] of Cefdinir RS taken
dihydrogen ethylenediamine tetraacetate TS.
Mobile phase B: To 500 mL of tetramethylammonium hy-
droxide TS (pH 5.5) add 300 mL of acetonitrile for liquid
length: 254 nm).
chromatography and 200 mL of methanol, and add 0.4 mL
of 0.1 mol/L disodium dihydrogen ethylenediamine tetraace-
tate TS.
409C.
Mobile phase: To 1000 mL of tetramethylammonium
Time after injection Mobile phase A Mobile phase B hydroxide TS (pH 5.5) add 0.4 mL of 0.1 mol/L disodium
of sample (min) (volz) (volz) dihydrogen ethylenediamine tetraacetate TS. To 900 mL of
0– 2 95 5 this solution add 60 mL of acetonitrile for liquid chromatog-
2 – 22 95 → 75 5 → 25 raphy and 40 mL of methanol.
22 – 32 75 → 50 25 → 50 Flow rate: Adjust so that the retention time of cefdinir is
32 – 37 50 50 about 8 minutes.
System performance: Dissolve 2 mg of Cefdinir RS and 5
Flow rate: 1.0 mL per minute (the retention time of mg of cefdinir lactam ring-cleavage lactones in 10 mL of 0.1
cefdinir is about 22 minutes). mol/L phosphate buffer solution, pH 7.0. When the proce-
Time span of measurement: For 37 minutes after injec- dure is run with 5 mL of this solution under the above operat-
JP XVII Official Monographs / Cefdinir Fine Granules 617
ing conditions, peak 1 and peak 2 of cefdinir lactam ring- (C14H13N5O5S2) in 1 capsule
cleavage lactones separated into 4 peaks, cefdinir, peak 3
and peak 4 of remaining cefdinir lactam ring-cleavage
lactones are eluted in this order. The resolution between the
Assay under Cefdinir.
peak 2 of cefdinir lactam ring-cleavage lactone and that of
cefdinir is not less than 1.2. The number of theoretical plates
and the symmetry factor of the peak of cefdinir are not less
than 2000 and not more than 1.5, respectively.
factor of the peak of cefdinir are not less than 2000 and not
conditions, the relative standard deviation of the peak areas
of cefdinir is not more than 1.0z.
Containers and storage Containers—Tight containers. ing conditions, the relative standard deviation of the peak
Storage—Light-resistant. area of cefdinir is not more than 1.0z.
Assay Weigh accurately not less than 5 Cefdinir Capsules,
take out the contents, and powder. Wash the empty capsules
Cefdinir Capsules with a little amount of diethyl ether, if necessary, allow to
stand at a room temperature to vaporize the adhering diethyl
セフジニルカプセル
ether, and weigh accurately the mass of the capsules to calcu-
late the mass of the contents. Weigh accurately an amount of
Cefdinir Capsules contain not less than 90.0z and the contents, equivalent to about 0.1 g (potency) of Cefdinir,
not more than 110.0z of the labeled potency of add 70 mL of 0.1 mol/L phosphate buffer solution (pH 7.0),
cefdinir (C14H13N5O5S2: 395.41). shake for 30 minutes, and add 0.1 mol/L phosphate buffer
solution (pH 7.0) to make exactly 100 mL. Centrifuge this
solution at 3000 revolutions per minute for 10 minutes, pipet
sules, with Cefdinir.
4 mL of the supernatant liquid, add 0.1 mol/L phosphate
Identification To an amount of the contents of Cefdinir buffer solution (pH 7.0) to make exactly 20 mL, and use this
Capsules, equivalent to 10 mg (potency) of Cefdinir, add 100 solution as the sample solution. Separately, weigh accurately
mL of 0.1 mol/L phosphate buffer solution (pH 7.0), ex- an amount of Cefdinir RS, equivalent to about 20 mg (po-
posure to ultrasonic waves for 1 minute, and filter. To 2 mL tency), dissolve in 0.1 mol/L phosphate buffer solution (pH
of the filtrate add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL, and use this solution as the
7.0) to make 20 mL, and determine the absorption spectrum standard solution. Proceed as directed in the Assay under
of this solution as directed under Ultraviolet-visible Spectro- Cefdinir.
Amount [mg (potency)] of cefdinir (C14H13N5O5S2)
225 nm and between 285 nm and 289 nm.
＝ MS × AT/AS × 5
MS: Amount [mg (potency)] of Cefdinir RS taken
sinker, using 900 mL of 2nd fluid for dissolution test as the
dissolution medium, the dissolution rate of a 50-mg capsule Cefdinir Fine Granules
in 30 minutes is not less than 80z, and that of a 100-mg cap-
セフジニル細粒
sule in 45 minutes is not less than 75z.
Start the test with 1 capsule of Cefdinir Capsules, with-
draw not less than 20 mL of the medium at the specified Cefdinir Fine Granules contain not less than 93.0z
minute after starting the test, and filter through a membrane and not more than 107.0z of the labeled potency of
filter with a pore size not exceeding 0.5 mm. Discard the first cefdinir (C14H13N5O5S2: 395.41).
add the dissolution medium to make exactly V? mL so that
ules, with Cefdinir.
each mL contains about 56 mg (potency) of Cefdinir, and use
this solution as the sample solution. Separately, weigh accu- Identification To an amount of Cefdinir Fine Granules,
rately about 28 mg (potency) of Cefdinir RS, and dissolve in equivalent to 10 mg (potency) of Cefdinir, add 100 mL of
the dissolution medium to make exactly 100 mL. Pipet 4 mL 0.1 mol/L phosphate buffer solution (pH 7.0), exposure to
of this solution, add the dissolution medium to make exactly ultrasonic waves for 1 minute, and filter. To 2 mL of the fil-
20 mL, and use this solution as the standard solution. Per- trate add 0.1 mol/L phosphate buffer solution (pH 7.0) to
form the test with exactly 20 mL each of the sample solution make 20 mL, and determine the absorption spectrum of this
and standard solution as directed under Liquid Chromatog- solution as directed under Ultraviolet-visible Spectropho-
raphy <2.01>, and determine the peak areas, AT and AS, of tometry <2.24>: it exhibits maxima between 221 nm and 225
cefdinir in each solution. nm and between 285 nm and 289 nm.
Dissolution rate (z) with respect to the labeled amount Uniformity of dosage units <6.02> The granules in single-
of cefdinir (C14H13N5O5S2) dose packages meet the requirement of the Mass variation
＝ MS × AT/AS × V?/V × 1/C × 180 test.
MS: Amount [mg (potency)] of Cefdinir RS taken Dissolution <6.10> When the test is performed at 50 revolu-
C: Labeled amount [mg (potency)] of cefdinir tions per minute according to the Paddle method, using 900
618 Cefditoren Pivoxil / Official Monographs JP XVII
mL of 2nd fluid for dissolution test as the dissolution me-
dium, the dissolution rate in 30 minutes of Cefdinir Fine Cefditoren Pivoxil
Granules is not less than 75z.
Start the test with an accurate amount of Cefdinir Fine セフジトレンピボキシル
Granules, equivalent to about 0.1 g (potency) of Cefdinir,
withdraw not less than about 20 mL of the medium at the
card the first 10 mL of the filtrate, and use the subsequent
filtrate as the sample solution. Separately, weigh accurately
about 28 mg (potency) of Cefdinir RS, and disslove in the
dissolution medium to make exactly 50 mL. Pipet 4 mL of
this solution, add the dissolution medium to make exactly 20
C25H28N6O7S3: 620.72
2,2-Dimethylpropanoyloxymethyl (6R,7R)-7-[(Z )-2-(2-
aminothiazol-4-yl)-2-(methoxyimino)acetylamino]-3-
<2.01>, and determine the peak areas, AT and AS, of cefdinir
[(1Z )-2-(4-methylthiazol-5-yl)ethenyl]-8-oxo-5-thia-1-
in each solution.
azabicyclo[4.2.0]oct-2-ene-2-carboxylate
Dissolution rate (z) with respect to the labeled amount [117467-28-4]
of cefdinir (C14H13N5O5S2)
＝ MS/MT × AT/AS × 1/C × 360 Cefditoren Pivoxil contains not less than 770 mg
MT: Amount (g) of Cefdinir Fine Granules taken
Cefditoren Pivoxil is expressed as mass (potency) of
C: Labeled amount [mg (potency)] of cefdinir
cefditoren (C19H18N6O5S3: 506.58).
(C14H13N5O5S2) in 1 g
Description Cefditoren Pivoxil occurs as a light yellowish
white to light yellow crystalline powder.
Assay under Cefdinir.
acetonitrile and in ethanol (95), very slightly sobuble in
diethyleter and practically insoluble in water.
ditions, the number of theoretical plates and the symmetry Identification (1) Dissolve 5 mg of Cefditoren Pivoxil in
factor of the peak of cefdinir are not less than 2000 and not 3 mL of hydroxylammonium chloride-ethanol TS, allow to
more than 2.0, respectively. stand for 5 minutes, add 1 mL of acidic ammonium iron
System repeatability: When the test is repeated 6 times (III) sulfate TS and shake: a red-brown color develops.
with 20 mL of the standard solution under the above operat- (2) Dissolve 1 mg of Cefditoren Pivoxil in 1 mL of dilute
ing conditions, the relative standard deviation of the peak hydrochloric acid and 4 mL of water, add 3 drops of sodium
area of cefdinir is not more than 1.0z. nitrite TS under ice-cooling, shake, and allow to stand for 2
minutes. Then add 1 mL of ammonium amidosulfate TS,
Assay Powder, if necessary, and weigh accurately an
shake well, and allow to stand for 1 minute, and add 1 mL
amount of Cefdinir Fine Granules, equivalent to about 0.1 g
of N, N-diethyl-N?-1-naphthylethylenediamine oxalate TS: a
(potency) of Cefdinir, add 70 mL of 0.1 mol/L phosphate
purple color develops.
buffer solution (pH 7.0), shake for 30 minutes, and add 0.1
mol/L phosphate buffer solution (pH 7.0) to make exactly
Cefditoren Pivoxil in methanol (1 in 50,000) as directed
100 mL. Centrifuge at 3000 revolutions per minute for 10
minutes, pipet 4 mL of the supernatant liquid, add 0.1
mol/L phosphate buffer solution (pH 7.0) to make 20 mL,
spectrum of a solution of Cefditoren Pivoxil RS prepared in
weigh accurately an amount of Cefdinir RS, equivalent to
about 20 mg (potency), dissolve in 0.1 mol/L phosphate
(4) Determine the 1H spectrum of a solution of Cefdito-
buffer solution (pH 7.0) to make exactly 100 mL, and use
ren Pivoxil in deuterated chloroform for nuclear magnetic
this solution as the standard solution. Proceed as directed in
resonance spectroscopy (1 in 50), using tetramethylsilane for
the Assay under Cefdinir.
nuclear magnetic resonance spectroscopy as an internal
Amount [mg (potency)] of cefdinir (C14H13N5O5S2) reference compound, as directed under Nuclear Magnetic
＝ M S × AT / AS × 5 Resonance Spectroscopy <2.21>: it exhibits single signals A,
B and C, at around d 1.1 ppm, at around d 2.4 ppm and at
around d 4.0 ppm, double signals D and E, at around d 6.4
Containers and storage Containers—Tight containers. ppm and at around d 6.7 ppm, and a single signal F at
Storage—Light-resistant. around d 8.6 ppm. The ratio of integrated intensity of each
signal A:B:C:D:E:F, is about 9:3:3:1:1:1.
Absorbance <2.24> E 11zcm (231 nm): 340 – 360 (50 mg, meth-
anol, 2500 mL).
D : －45 – －529(50 mg, metha-
nol, 10 mL, 100 mm).
JP XVII Official Monographs / Cefditoren Pivoxil Fine Granules 619

Cefditoren Pivoxil according to Method 2, and perform the Cefditoren Pivoxil Fine Granules
Lead Solution (not more than 10 ppm). セフジトレンピボキシル細粒
Cefditoren Pivoxil Fine Granules contain not less
Water <2.48> Not more than 1.5z (0.5 g, volumetric titra- than 90.0z and not more than 110.0z of the labeled
tion, direct titration). potency of cefditoren (C19H18N6O5S3: 506.58).
Residue on ignition Being specified separately when the Method of preparation Prepare as directed under Gran-
drug is granted approval based on the Law. ules, with Cefditoren Pivoxil.
Assay Conduct this procedure using light-resistant vessels. Identification To an amount of powdered Cefditoren
Weigh accurately an amount of Cefditoren Pivoxil and Cef- Pivoxil Fine Granules, equivalent to 0.1 g (potency) of Cef-
ditoren Pivoxil RS, equivalent to about 40 mg (potency), dis- ditoren Pivoxil, add 10 mL of acetonitrile, shake vigorously,
solve in 40 mL of acetonitrile, add exactly 10 mL each of the and filter. To 1 mL of the filtrate add acetonitrile to make 50
internal standard solution, and add acetonitrile to make 100 mL. To 1 mL of this solution add acetonitrile to make 20
mL, and use these solutions as the sample solution and mL, and determine the absorption spectrum of this solution
standard solution. Perform the test with 10 mL each of the as directed under Ultraviolet-visible Spectrophotometry
sample solution and standard solution as directed under Liq- <2.24>: it exhibits a maximum between 230 nm and 234 nm.
Purity Related substances—Being specified separately
when the drug is granted approval based on the Law.
of cefditoren pivoxil to that of the internal standard.
Amount [ mg (potency)] of cefditoren (C19H18N6O5S3)
pressure not exceeding 0.67 kPa, 609
C, 3 hours).
＝ MS × QT/QS × 1000
MS: Amount [mg (potency)] of Cefditoren Pivoxil RS
taken
test.
Internal standard solution—A solution of propyl p-hydrox-
ybenzoate in acetonitrile (1 in 200).
900 mL of 1st fluid for dissolution test as the dissolution
medium, the dissolution rate in 15 minutes of Cefditoren
length: 230 nm).
Pivoxil Fine Granules is not less than 80z.
Start the test with an accurately weighed amount of Cef-
ditoren Pivoxil Fine Granules, equivalent to about 0.1 g (po-
tency) of Cefditoren Pivoxil, withdraw not less than 20 mL
259 C.
Mobile phase: Dissolve 1.58 g of ammonium formate in
900 mL of water, adjust to pH 6.0 with diluted formic acid
pipet 2 mL of the subsequent filtrate, add water to make
(1 in 250), and add water to make 1000 mL. To 450 mL of
this solution add 275 mL of acetonitrile and 275 mL of
Separately, weigh accurately an amount of Cefditoren
methanol.
Pivoxil RS, equivalent to about 22 mg (potency), dissolve in
Flow rate: Adjust so that the retention time of cefditoren
20 mL of diluted acetonitrile (3 in 4), and add the dissolution
pivoxil is about 15 minutes.
medium to make exactly 200 mL. Pipet 2 mL of this solu-
as the standard solution. Determine the absorbances, AT and
ditions, the internal standard and cefditoren pivoxil are
<2.24>, using water as the control.
System repeatability: When the test is repeated 5 times Dissolution rate (z) with respect to the labeled amount
with 10 mL of the standard solution under the above operat- of cefditoren pivoxil (C25H28N6O7S3)
ing conditions, the relative standard deviation of the ratios ＝ MS/MT × AT/AS × 1/C × 450
of the peak area of cefditoren pivoxil to that of the internal
MS: Amount [mg(potency)] of Cefditoren Pivoxil RS
taken
Containers and storage Containers—Tight containers. MT: Amount (g) of Cefditoren Pivoxil Fine Granules
Storage—Light-resistant. taken
C: Labeled amount [mg(potency)] of cefditoren pivoxil
(C25H28N6O7S3) in 1 g
Assay Conduct this procedure using light-resistant vessels.
Weigh accurately an amount of powdered Cefditoren Pivoxil
Fine Granules, equivalent to about 40 mg (potency) of Cef-
ditoren Pivoxil, add 70 mL of diluted acetonitrile (3 in 4),
and shake vigorously. To this solution add exactly 10 mL of
the internal standard solution, then add acetonitrile to make
620 Cefditoren Pivoxil Tablets / Official Monographs JP XVII
100 mL, filter, and use the filtrate as the sample solution. Dissolution <6.10> When the test is performed at 50 revolu-
Separately, weigh accurately an amount of Cefditoren tions per minute according to the Paddle method, using
Pivoxil RS, equivalent to about 20 mg (potency), dissolve in 900 mL of 1st fluid for dissolution test as the dissolution
20 mL of acetonitrile, add exactly 5 mL of the internal medium, the dissolution rate in 20 minutes of Cefditoren
standard solution, then add acetonitrile to make 50 mL, and Pivoxil Tablets is not less than 85z.
use this solution as the standard solution. Proceed as di- Start the test with 1 tablet of Cefditoren Pivoxil Tablets,
rected in the Assay under Cefditoren Pivoxil. withdraw not less than 20 mL of the medium at the specified
Amount [mg (potency)] of cefditoren (C19H18N6O5S3)
＝ M S × QT / QS × 2
MS: Amount [mg (potency)] of Cefditoren Pivoxil RS filtrate, add water to make exactly V? mL so that each mL
taken contains about 11 mg (potency) of Cefditoren Pivoxil, and
use this solution as the sample solution. Separately, weigh
accurately an amount of Cefditoren Pivoxil RS, equivalent
droxybenzoate in acetonitrile (1 in 200).
to about 22 mg (potency), dissolve in 20 mL of diluted aceto-
Containers and storage Containers—Tight containers. nitrile (3 in 4), then add the dissolution medium to make ex-
Storage—Light-resistant. actly 200 mL. Pipet 2 mL of this solution, add water to make
exactly 20 mL, and use this solution as the standard solution.
Determine the absorbances, AT and AS, at 272 nm of the
Cefditoren Pivoxil Tablets sample solution and standard solution as directed under
Ultraviolelt-visible Spectrophotometry <2.24> using water as
セフジトレンピボキシル錠 the control.
Cefditoren Pivoxil Tablets contain not less than of cefditoren pivoxil (C25H28N6O7S3)
90.0z and not more than 110.0z of the labeled ＝ MS × AT/AS × V?/V × 1/C × 45
potency of cefditoren (C19H18N6O5S3: 506.58).
Method of preparation Prepare as directed under Tablets, taken
with Cefditoren Pivoxil. C: Labeled amount [mg (potency)] of cefditoren pivoxil
(C25H28N6O7S3) in 1 tablet
Identification To an amount of powdered Cefditoren
Pivoxil Tablets, equivalent to 35 mg (potency) of Cefditoren Assay Conduct this procedure using light-resistant vessels.
Pivoxil, add 100 mL of methanol, shake, and filter. To 5 mL To an amount of Cefditoren Pivoxil Tablets, equivalent to
of the filtrate add methanol to make 100 mL, and determine 0.5 g (potency) of Cefditoren Pivoxil, add 63 mL of the 1st
the absorption spectrum of this solution as directed under fluid for disintegration test, shake vigorously, add about 125
Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a mL of acetonitrile, shake again, and add acetonitrile to
maximum between 229 nm and 233 nm. make exactly 250 mL. Pipet 10 mL of this solution, add ex-
actly 5 mL of the internal standard solution, then add
Purity Related substances—Being specified separately
diluted acetonitrile (3 in 4) to make 50 mL, filter, and use the
when the drug is granted approval based on the Law.
filtrate as the sample solution. Separately, weigh accurately
Loss on drying <2.41> Not more than 4.0z (0.5 g, reduced an amount of Cefditoren Pivoxil RS, equivalent to about 20
pressure not exceeding 0.67 kPa, 609C, 3 hours). mg (potency), dissolve in 20 mL of acetonitrile, add exactly 5
mL of the internal standard solution, then add acetonitrile to
Proceed as directed in the Assay under Cefditoren Pivoxil.
Conduct this procedure using light-resistant vessels. To 1 Amount [mg (potency)] of cefditoren (C19H18N6O5S3)
tablet of Cefditoren Pivoxil Tablets add 12.5 mL of the 1st ＝ MS × QT/QS × 25
fluid for disintegration test, shake vigorously, add about 25
mL of acetonitrile, shake again, and add acetonitrile to
taken
make exactly 50 mL. Pipet V mL of this solution, equivalent
to about 20 mg (potency) of Cefditoren Pivoxil, add exactly Internal standard solution—A solution of propyl parahy-
5 mL of the internal standard solution, then add diluted droxybenzoate in acetonitrile (1 in 200).
acetonitrile (3 in 4) to make 50 mL, filter, and use the filtrate
as the sample solution. Separately, weigh accurately an
amount of Cefditoren Pivoxil RS, equivalent to about 20 mg
(potency), dissolve in 20 mL of acetonitrile, add exactly 5
mL of the internal standard solution, then add acetonitrile to
Proceed as directed in the Assay under Cefditoren Pivoxil.
Amount [mg (potency)] of cefditoren (C19H18N6O5S3)
＝ MS × QT/QS × 50/V
taken
Internal standard solution—A solution of propyl para-
hydroxybezoate in acetonitrile (1 in 200).
JP XVII Official Monographs / Cefepime Dihydrochloride Hydrate 621
drate in 10 mL of water: the pH of this solution is between

Cefepime Dihydrochloride Hydrate 1.6 and 2.1.
セフェピム塩酸塩水和物
of Cefepime Dihydrochloride Hydrate in 5 mL of a solution
of L-arginine (3 in 50): the solution is clear and has no more
color than Matching Fluid H.
Cefepime Dihydrochloride Hydrate according to Method 2,
and perform the test. Prepare the control solution with 2.0
mL of Standard Lead Solution (not more than 20 ppm).
(3) N-Methylpyrrolidine—Weigh accurately an amount
C19H24N6O5S2.2HCl.H2O: 571.50
of Cefepime Dihydrochloride Hydrate equivalent to about
(6R,7R)-7-[(Z )-2-(2-Aminothiazol-4-yl)-2-
80 mg (potency), dissolve in diluted nitric acid (2 in 3125) to
(methoxyimino)acetylamino]-3-(1-methylpyrrolidinium-1-
make exactly 10 mL, and use this solution as the sample so-
ylmethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
lution. Separately, put 30 mL of water in a 100-mL volumet-
carboxylate dihydrochloride monohydrate
ric flask, weigh accurately the mass of flask, then add about
[123171-59-5]
0.125 g of N-methylpyrrolidine, weigh accurately the mass of
the flask again, and add water to make exactly 100 mL.
Cefepime Dihydrochloride Hydrate contains not
Pipet 4 mL of this solution, add diluted nitric acid (2 in
less than 835 mg (potency) and not more than 886 mg
3125) to make exactly 100 mL, and use this solution as the
(potency) per mg, calculated on the anhydrous
basis. The potency of Cefepime Dihydrochloride
Hydrate is expressed as mass (potency) of cefepime
(C19H24N6O5S2: 480.56).
lowing conditions, and determine the peak areas, AT and
Description Cefepime Dihydrochloride Hydrate occurs as a AS, of N-methylpyrrolidine by the automatic integration
white to yellowish white, crystals or crystalline powder. method. Calculate the amount of N-methylpyrrolidine per
It is freely soluble in water and in methanol, and slightly 1 mg (potency) of Cefepime Dihydrochloride Hydrate by the
soluble in ethanol (95), and practically in soluble in diethyl following equation: not more than 0.5z. The sample solu-
ether. tion must be tested within 20 minutes after preparation.
Identification (1) Dissolve 0.02 g of Cefepime Dihydro- Amount (z) of N-methylpyrrolidine
chloride Hydrate in 2 mL of water, add 1 mL of a solution ＝ (MS × f )/MT × AT/AS × 1/250
of hydroxylammonium chloride (1 in 10) and 2 mL of so-
MS: Amount (mg) of N-methylpyrrolidine taken
dium hydroxide TS, allow to stand for 5 minutes, then add 3
MT: Amount [mg (potency)] of Cefepime Dihydrochloride
mL of 1 mol/L hydrochloric acid TS and 3 drops of iron
Hydrate taken
(III) chloride TS: a red-brown color develops.
f: Purity (z) of N-methylpyrrolidine
(2) Determine the absorption spectra of solutions (1 in
20,000) of Cefepime Dihydrochloride Hydrate and Cefepime Operating conditions—
Dihydrochloride RS as directed under Ultraviolet-visible Detector: An electric conductivity detector.
Spectrophotometry <2.24>, and compare these spectra: both Column: A plastic tube 4.6 mm in inside diameter and 5
spectra exhibit similar intensities of absorption at the same cm in length, packed with hydrophilic silica gel for liquid
wavelengths. chromatography carrying sulfonic acid groups having the
(3) Determine the infrared absorption spectra of exchange capacity of about 0.3 meq per g (5 mm in particle
Cefepime Dihydrochloride Hydrate and Cefepime Dihydro- diameter).
chloride RS as directed in the potassium bromide disk Column temperature: A constant temperature of about
method under Infrared Spectrophotometry <2.25>, and com- 359C.
pare these spectra: both spectra exhibit similar intensities of Mobile phase: To 990 mL of diluted nitric acid (2 in 3125)
absorption at the same wave numbers. add 10 mL of acetonitrile.
(4) Determine the 1H spectrum of a solution of Cefepime Flow rate: 1.0 mL per minute.
Dihydrochloride Hydrate in heavy water for nuclear mag- System suitability—
netic resonance spectroscopy (1 in 10) as directed under System performance: To 20 mL of a solution of sodium
Nuclear Magnetic Resonance Spectroscopy <2.21>, using chloride (3 in 1000) add 0.125 g of N-methylpyrrolidine, and
sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic add water to make 100 mL. To 4 mL of this solution add
resonance spectroscopy as an internal reference compound: diluted nitric acid (2 in 3125) to make 100 mL. When the
it exhibits single signals, A and B, at around d 3.1 ppm and procedure is run with 100 mL of this solution under the
at around d 7.2 ppm, respectively, and the ratio of integrated above operating conditions, sodium and N-methylpyrroli-
intensity of each signal, A:B, is about 3:1. dine are eluted in this order with the resolution between these
(5) Dissolve 15 mg of Cefepime Dihydrochloride Hy- peaks being not less than 2.0.
drate in 5 mL of water, and add 2 drops of silver nitrate TS: System repeatability: When the test is repeated 5 times
a white turbidity is produced. with 100 mL of the standard solution under the above operat-
Absorbance <2.24> E 11zcm (259 nm): 310 – 340 (50 mg calcu-
areas of N-methylpyrrolidine is not more than 4.0z.
lated on the anhydrous basis, water, 1000 mL).
(4) Related substances—Dissolve about 0.1 g of
D : ＋39 – ＋479 (60 mg calcu- Cefepime Dihydrochloride Hydrate in the mobile phase A to
lated on the anhydrous basis, water, 20 mL, 100 mm). make 50 mL, and use this solution as the sample solution.
Perform the test with 5 mL of the sample solution as directed
pH <2.54> Dissolve 0.1 g of Cefepime Dihydrochloride Hy-
622 Cefepime Dihydrochloride for Injection / Official Monographs JP XVII
lowing conditions, and determine the area of each peak by Amount [ mg (potency)] of cefepime (C19H24N6O5S2)
the automatic integration method. Calculate the total ＝ MS × AT/AS × 1000
amount of the peaks other than cefepime by the area percen-
MS: Amount [mg (potency)] of Cefepime Dihydrochloride
tage method: not more than 0.5z.
RS taken
Detector: An ultraviolet absorption photometer (wave- Operating conditions—
length: 254 nm). Detector: An ultraviolet absorption photometer (wave-
Column: A stainless steel column 4.6 mm in inside diame- length: 254 nm).
ter and 25 cm in length, packed with octadecylsilanized silica Column: A stainless steel column 3.9 mm in inside diame-
gel for liquid chromatography (10 mm in particle diameter). ter and 30 cm in length, packed with octadecylsilanized silica
Column temperature: A constant temperature of about gel for liquid chromatography (10 mm in particle diameter).
259 C. Column temperature: A constant temperature of about
Mobile phase A: Dissolve 0.57 g of ammonium dihydro- 409C.
genphosphate in 1000 mL of water. Mobile phase: Adjust a solution of sodium 1-pentanesul-
Mobile phase B: Acetonitrile. fonate (261 in 100,000) to pH 3.4 with acetic acid (100), then
Flowing of mobile phase: Control the gradient by mixing adjust this solution to pH 4.0 with a solution of potassium
the mobile phases A and B as directed in the following table. hydroxide (13 in 20). To 950 mL of this solution add 50 mL
of acetonitrile.
Time after injection Mobile phase A Mobile phase B Flow rate: Adjust so that the retention time of cefepime is
of the sample (min) (volz) (volz) about 8 minutes.
0 – 25 100 → 75 0 → 25 System performance: When the procedure is run with 10
Flow rate: Adjust so that the retention time of cefepime is ditions, the number of theoretical plates of the peak of
about 9.5 minutes. cefepime is not less than 1500.
Time span of measurement: About 2.5 times as long as the System repeatability: When the test is repeated 5 times
retention time of cefepime. with 10 mL of the standard solution under the above operat-
System suitability— ing conditions, the relative standard deviation of the peak
Test for required detectability: To 1 mL of the sample so- areas of cefepime is not more than 2.0z.
lution add the mobile phase A to make 10 mL, and use this Containers and storage Containers—Hermetic containers.
solution as the solution for system suitability test. To 1 mL Storage—Light-resistant.
of the solution for system suitability test add the mobile
phase A to make 10 mL, and use this solution as the solution
for test for required detectability. Pipet 1 mL of the solution
for test for required detectability, add the mobile phase A to
Cefepime Dihydrochloride for
make exactly 10 mL. Conform that the peak area of Injection
cefepime obtained from 5 mL of this solution is equivalent to
7 to 13z of that of cefepime obtained from 5 mL of the solu- 注射用セフェピム塩酸塩
tion for test for required detectability.
System performance: When the procedure is run with 5 mL Cefepime Dihydrochloride for Injection is a prepa-
of the solution for system suitability test under the above ration for injection, which is dissolved before use.
operating conditions, the number of theoretical plates of the It contains not less than 95.0z and not more
peak of cefepime is not less than 6000. than 110.0z of the labeled potency of cefepime
System repeatability: When the test is repeated 3 times (C19H24N6O5S2: 480.56).
above operating conditions, the relative standard deviation Method of preparation Prepare as directed under Injec-
of the peak areas of cefepime is not more than 2.0z. tions, with Cefepime Dihydrochloride Hydrate.
Water <2.48> Not less than 3.0z and not more than 4.5z Description Cefepime Dihydrochloride for Injection occurs
(Weigh accurately about 50 mg of Cefepime Dihydrochlo- as a white to pale yellow powder.
ride Hydrate, dissolve in exactly 2 mL of methanol for water Identification (1) Dissolve 40 mg of Cefepime Dihydro-
determination and perform the test with exactly 0.5 mL of chloride in 2 mL of water, add 1 mL of a solution of hydrox-
this solution; coulometric titration). ylammonium chloride (1 in 10) and 2 mL of sodium hydrox-
Residue on ignition <2.44> Not more than 0.1z (1 g). ide TS, allow to stand for 5 minutes, then add 3 mL of 1
mol/L hydrochloric acid TS and 3 drops of iron (III) chlo-
Bacterial endotoxins <4.01> Less than 0.04 EU/mg (po- ride TS: a red-brown color develops.
tency). (2) Determine the absorption spectrum of a solution of
Assay Weigh accurately an amount of Cefepime Dihydro- Cefepime Dihydrochloride for Injection (1 in 12,500) as di-
chloride Hydrate and Cefepime Dihydrochloride RS, equiva- rected under Ultraviolet-visible Spectrophotometry <2.24>: it
lent to about 60 mg (potency), dissolve in the mobile phase exhibits maxima between 233 nm and 237 nm and between
to make exactly 50 mL, and use these solutions as the sample 255 nm and 259 nm.
solution and the standard solution, respectively. Perform the pH <2.54> The pH of a solution obtained by dissolving an
test with exactly 10 mL each of the sample solution and amount of Cefepime Dihydrochloride for Injection, equiva-
standard solution as directed under Liquid Chromatography lent to 0.5 g (potency) of Cefepime Dihydrochloride Hy-
<2.01> according to the following conditions, and determine drate, in 5 mL of water is between 4.0 and 6.0.
the peak areas, AT and AS, of cefepime in each solution.
JP XVII Official Monographs / Cefixime Hydrate 623
amount of Cefepime Dihydrochloride for Injection, equiva- Amount [ mg (potency)] of cefepime (C19H24N6O5S2)
lent to 0.5 g (potency) of Cefepime Dihydrochloride Hy- ＝ MS × AT/AS × 1000
drate, in 5 mL of water: the solution is clear and colorless or
MS: Amount [mg (potency)] of Cefepime Dihydrochloride
light yellow. The color is not darker than Matching Fluid I.
RS taken
(2) N-Methylpyrrolidine—Weigh accurately an amount
of Cefepime Dihydrochloride for Injection, equivalent to Containers and storage Containers—Hermetic containers.
about 0.2 g (potency) of Cefepime Dihydrochloride Hydrate, Storage—Light-resistant.
dissolve in diluted nitric acid (2 in 625) to make exactly 20
transfer 30 mL of water into a 100-mL volumetric flask, Cefixime Hydrate
weigh accurately the mass of the flask, add about 0.125 g of
N-methylpyrrolidine, then weigh accurately the mass, and セフィキシム水和物
add water to make exactly 100 mL. Pipet 4 mL of this solu-
tion, add diluted nitric acid (2 in 3125) to make exactly 100
the peak areas of N-methylpyrrolidine, AT and AS, by the
automatic integration method within 20 minutes after the C16H15N5O7S2.3H2O: 507.50
sample solution is prepared. Calculate the amount of N- (6R,7R)-7-[(Z )-2-(2-Aminothiazol-4-yl)-2-
methylpyrrolidine per mg (potency) of Cefepime Dihydro- (carboxymethoxyimino)acetylamino]-8-oxo-3-vinyl-5-thia-1-
chloride for Injection by the following formula: not more azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid trihydrate
than 1.0z. [125110-14-7]
Amount (z) of N-methylpyrrolidine
＝ (MS × f )/MT × AT/AS × 1/125
Cefixime Hydrate contains not less than 930 mg
MS: Amount (mg) of N-methylpyrrolidine taken mg, calculated on the anhydrous basis. The potency of
MT: Amount [mg (potency)] of Cefepime Dihydrochloride Cefixime Hydrate is expressed as mass (potency) of
for Injection taken cefixime (C16H15N5O7S2: 453.45).
f: Purity (z) of N-methylpyrrolidine
Description Cefixime Hydrate occurs as a white to light
Operating conditions— yellow crystalline powder.
Proceed as directed in the operating conditions in the It is freely soluble in methanol and in dimethylsulfoxide,
Purity (3) under Cefepime Dihydrochloride Hydrate. sparingly soluble in ethanol (99.5), and practically insoluble
System suitability— in water.
(3) under Cefepime Dihydrochloride Hydrate.
solution of Cefixime Hydrate in 0.1 mol/L phosphate buffer
Water <2.48> Not more than 4.0z (Weigh accurately about solution (pH 7.0) (1 in 62,500) as directed under Ultraviolet-
50 mg of Cefepime Dihydrochloride for Injection, dissolve visible Spectrophotometry <2.24>, and compare the spectrum
in exactly 2 mL of methanol for water determination, and with the Reference Spectrum or the spectrum of a solution of
perform the test with exactly 0.5 mL of this solution: coulo- Cefixime RS prepared in the same manner as the sample so-
metric titration). lution: both spectra exhibit similar intensities of absorption
tency).
Cefixime Hydrate as directed in the potassium bromide disk
Uniformity of dosage units <6.02> It meets the requirement method under Infrared Spectrophotometry <2.25>, and com-
of the Mass variation test. pare the spectrum with the Reference Spectrum or the spec-
trum of Cefixime RS: both spectra exhibit similar intensities
(3) Dissolve 0.05 g of Cefixime Hydrate in 0.5 mL of a
Insoluble particulate matter <6.07> It meets the require- mixture of deuterated dimethylsulfoxide for nuclear mag-
ment. netic resonance spectroscopy and heavy water for nuclear
magnetic resonance spectroscopy (4:1). Determine the 1H
spectrum of this solution, using tetramethylsilane for nuclear
magnetic resonance spectroscopy as an internal reference
Assay Weigh accurately the mass of the contents of not less compound, as directed under Nuclear Magnetic Resonance
than 10 Cefepime Dihydrochloride for Injection. Weigh ac- Spectroscopy <2.21>: it exhibits a single signal A at around
curately an amount of the content, equivalent to about 60 d 4.7 ppm, and a multiple signal B between d 6.5 ppm and
mg (potency) of Cefepime Dihydrochloride Hydrate, dis- d 7.4 ppm. The ratio of integrated intensity of these signals,
solve in the mobile phase to make exactly 50 mL, and use A:B, is about 1:1.
Optical rotation <2.49> [a]20D : －75 – －889 (0.45 g calcu-
rately an amount of Cefepime Dihydrochloride RS, equiva-
lated on the anhydrous bases, a solution of sodium hydrogen
lent to about 60 mg (potency), dissolve in the mobile phase
carbonate (1 in 50), 50 mL, 100 mm).
solution. Proceed as directed in the Assay under Cefepime Purity Dissolve 0.1 g of Cefixime Hydrate in 100 mL of 0.1
Dihydrochloride Hydrate. mol/L phosphate buffer solution (pH 7.0), and use this solu-
624 Cefixime Capsules / Official Monographs JP XVII
tion as the sample solution. Perform the test with 10 mL of System suitability—
the sample solution as directed under Liquid Chromatogra- System performance: When the procedure is run with 10
phy <2.01> according to the following conditions, determine mL of the standard solution under the above operating con-
the areas of the peaks by the automatic integration method, ditions, the number of theoretical plates and the symmetry
and calculate the amounts of these peak areas by the area factor of the peak of cefixime are not less than 4000 and not
percentage method: the amount of each peak other than more than 2.0, respectively.
cefixime is not more than 1.0z, and the total amount of the System repeatability: When the test is repeated 6 times
peaks other than cefixime is not more than 2.5z. with 10 mL of the standard solution under the above operat-
Operating conditions— ing conditions, the relative standard deviation of peak areas
Detector, column, column temperature, mobile phase, and of cefixime is not more than 2.0z.
the Assay.
retention time of cefixime beginning after the solvent peak.
Test for required detectability: Pipet 1 mL of the sample Cefixime Capsules
セフィキシムカプセル
7.0) to make exactly 100 mL. Confirm that the peak height
of cefixime obtained from 10 mL of this solution is equiva-
lent to 20 to 60 mm. Cefixime Capsules contain not less than 90.0z and
System performance: Dissolve about 2 mg of Cefixime RS not more than 105.0z of the labeled potency of
in 200 mL of 0.1 mol/L phosphate buffer solution (pH 7.0) cefixime (C16H15N5O7S2: 453.45).
and use this solution as the solution for system suitability
test. When the procedure is run with 10 mL of the solution
sules, with Cefixime Hydrate.
according to the above operating conditions, the number of
theoretical plates and the symmetry factor of the peak of Identification Take out the contents of Cefixime Capsules,
cefixime are not less than 4000 and not more than 2.0, to a quantity of the contents of Cefixime Capsules, equiva-
respectively. lent to 70 mg (potency) of Cefixime Hydrate, add 100 mL of
System repeatability: When the test is repeated 6 times 0.1 mol/L phosphate buffer solution (pH 7.0), shake for 30
with 10 mL of the solution for system suitability test under minutes, and filter. To 1 mL of the filtrate add 0.1 mol/L
the above operating conditions, the relative standard devia- phosphate buffer solution (pH 7.0) to make 50 mL. Deter-
tion of the peak areas of cefixime is not more than 2.0z. mine the absorption spectrum of this solution as directed
under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-
Water <2.48> Not less than 9.0 and not more than 12.0z
its a maximum between 286 nm and 290 nm.
Purity Related substances—Take out the contents of
Cefixime Capsules, to a quantity of the contents of Cefixime
Assay Weigh accurately an amount of Cefixime Hydrate Capsules, equivalent to 0.1 g (potency) of Cefixime Hydrate,
and Cefixime RS, equivalent to about 0.1 g (potency), and add 100 mL of 0.1 mol/L phosphate buffer solution (pH
dissolve in 0.1 mol/L phosphate buffer solution (pH 7.0) to 7.0), shake for 30 minutes, filter, and use the filtrate as the
make exactly 100 mL each. Pipet 10 mL each of these solu- sample solution. Perform the test with 10 mL of the sample
tions, add 0.1 mol/L phosphate buffer solution (pH 7.0) to solution as directed under Liquid Chromatography <2.01>
make exactly 50 mL each, and use these solutions as the sam- according to the following conditions. Determine each peak
ple solution and the standard solution, respectively. Perform area from the sample solution by the automatic integration
the test with exactly 10 mL each of the sample solution and method, and calculate the amount of them by the area per-
standard solution as directed under Liquid Chromatography centage method: the amount of each peak other than
<2.01> according to the following conditions, and determine cefixime is not more than 1.0z, and the total amount of the
the peak areas, AT and AS, of cefixime in each solution. peaks other than cefixime is not more than 2.5z.
Amount [ mg (potency)] of C16H15N5O7S2
Detector, column, column temperature, mobile phase and
＝ MS × AT/AS × 10000
MS: Amount [mg (potency)] of Cefixime RS taken the Assay under Cefixime Hydrate.
Time span for measurement: Proceed as directed in the
operating conditions in the Purity under Cefixime Hydrate.
length: 254 nm).
Test for required detectability: Pipet 1 mL of the sample
and 125 mm in length, packed with octadecylsilanized silica
7.0) to make exactly 100 mL, and use this solution as the so-
lution for system suitability test. Pipet 1 mL of the solution
for system suitability test, and add 0.1 mol/L phosphate
409 C.
buffer solution (pH 7.0) to make exactly 10 mL. Confirm
Mobile phase: To 25 mL of a solution of tetrabutylammo-
that the peak area of cefixime obtained from 10 mL of this
nium hydroxide TS (10 in 13) add water to make 1000 mL,
solution is equivalent to 7 to 13z of that obtained from 10
adjust to pH 6.5 with diluted phosphoric acid (1 in 10). To
mL of the solution for system suitability test.
Flow rate: Adjust so that the retention time of cefixime is
about 10 minutes.
JP XVII Official Monographs / Cefmenoxime Hydrochloride 625
the symmetry factor of the peak of cefixime are not less than mL of the standard solution under the above operating con-
4000 and not more than 2.0, respectively. ditions, the number of theoretical plates and the symmetry
System repeatability: When the test is repeated 6 times factor of the peak of cefixime are not less than 4000 and not
with 10 mL of the solution for system suitability test under more than 2.0, respectively.
the above operating conditions, the relative standard devia- System repeatability: When the test is repeated 6 times
tion of the peak area of cefixime is not more than 2.0z. with 20 mL of the standard solution under the above operat-
Water <2.48> Not more than 12.0z (0.1 g of the contents,
area of cefixime is not more than 2.0z.
volumetric titration, direct titration).
Assay Take out the contents of not less than 20 Cefixime
Capsules, weigh accurately the mass of the contents, and
powder. Weigh accurately a portion of the powder, equiva-
lent to about 0.1 g (potency) of Cefixime Hydrate, add 70
Take out the contents of 1 capsule of Cefixime Capsules,
mL of 0.1 mol/L phosphate buffer solution (pH 7.0) and
and to the contents and the capsule shells add 7V/10 mL of
shake for 30 minutes, add 0.1 mol/L phosphate buffer solu-
0.1 mol/L phosphate buffer solution (pH 7.0), shake for 30
tion (pH 7.0) to make exactly 100 mL. Centrifuge this solu-
minutes, and add 0.1 mol/L phosphate buffer solution (pH
tion, pipet 10 mL of the supernatant liquid, add 0.1 mol/L
7.0) to make exactly V mL so that each mL contains about
1 mg (potency) of Cefixime Hydrate. Centrifuge this solu-
tion, pipet 10 mL of the supernatant liquid, add 0.1 mol/L
weigh accurately an amount of Cefixime RS, equivalent to
weigh accurately an amount of Cefixime RS, equivalent to
this solution as the standard solution. Then, proceed as di-
rected in the Assay under Cefixime Hydrate.
this solution as the standard solution. Then, proceed as Amount [mg (potency)] of cefixime (C16H15N5O7S2)
directed in the Assay under Cefixime Hydrate. ＝ M S × AT / AS × 5
Amount [mg (potency)] of cefixime (C16H15N5O7S2) MS: Amount [mg (potency)] of Cefixime RS taken
＝ MS × AT/AS × V/20
MS: Amount [mg (potency)] of Cefixime RS taken
tions per minute according to the Paddle method using the Cefmenoxime Hydrochloride
sinker, using 900 mL of disodium hydrogen phosphate-citric
セフメノキシム塩酸塩
acid buffer solution (pH 7.5) as the dissolution medium, the
dissolution rates in 60 minutes of 50-mg (potency) capsule
and in 90 minutes of 100-mg (potency) capsule are not less
than 80z, respectively.
Start the test with 1 capsule of Cefixime Capsules, with-
(C16H17N9O5S3.)2.HCl: 1059.58
add the dissolution medium to make exactly V? mL so that
each mL contains about 56 mg (potency) of Cefixime Hy-
(methoxyimino)acetylamino]-3-(1-methyl-1H-tetrazol-5-
drate, and use this solution as the sample solution. Sepa-
ylsulfanylmethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-
rately, weigh accurately an amount of Cefixime RS, equiva-
ene-2-carboxylic acid hemihydrochloride
lent to about 28 mg (potency), and dissolve in the dissolution
[75738-58-8]
medium to make exactly 100 mL. Pipet 4 mL of this solu-
tion, add the dissolution medium to make exactly 20 mL,
Cefmenoxime Hydrochloride contains not less than
890 mg (potency) and not more than 975 mg (potency)
per mg, calculated on the anhydrous basis. The
potency of Cefmenoxime Hydrochloride is expressed
as mass (potency) of cefmenoxime (C16H17N9O5S3:
the peak areas, AT and AS, of cefixime in each solution.
511.56).
Description Cefmenoxime Hydrochloride occurs as white
of cefixime (C16H15N5O7S2)
to light orange-yellow, crystals or crystalline powder.
＝ MS × AT/AS × V?/V × 1/C × 180
It is freely soluble in formamide and in dimethylsulfoxide,
MS: Amount [mg (potency)] of Cefixime RS taken slightly soluble in methanol, very slightly soluble in water,
C: Labeled amount [mg (potency)] of Cefixime Hydrate in and practically insoluble in ethanol (95).
1 capsule
Operating conditions— solution of Cefmenoxime Hydrochloride in 0.1 mol/L phos-
Proceed as directed in the operating conditions in the phate buffer solution (pH 6.8) (3 in 200,000) as directed
Assay under Cefixime Hydrate. under Ultraviolet-visible Spectrophotometry <2.24>, and
System suitability— compare the spectrum with the Reference Spectrum or the
System performance: When the procedure is run with 20 spectrum of a solution of Cefmenoxime Hydrochloride RS
626 Cefmenoxime Hydrochloride / Official Monographs JP XVII
prepared in the same manner as the sample solution: both Amount (z) of 1-methyl-1H-tetrazol-5-thiol
spectra exhibit similar intensities of absorption at the same ＝ MSa/MT × ATa/ASa × 20
wavelengths.
Amount (z) of total related substances
(2) Determine the infrared absorption spectrum of Cef-
＝ MSa/MT × ATa/ASa × 20
menoxime Hydrochloride as directed in the potassium bro-
＋ MSb/MT × ST/ASb × 5
and compare the spectrum with the Reference Spectrum or MSa: Amount (g) of 1-methyl-1H-tetrazol-5-thiol taken
the spectrum of Cefmenoxime Hydrochloride RS: both spec- MSb: Amount (g) of Cefmenoxime Hydrochloride RS
tra exhibit similar intensities of absorption at the same wave taken
numbers. MT: Amount (g) of Cefmenoxime Hydrochloride taken
(3) Determine the 1H spectrum of a solution of Cef- ASa: Peak area of 1-methyl-1H-tetrazol-5-thiol from the
menoxime Hydrochloride in deuterated dimethylsulfoxide standard solution (1)
for nuclear magnetic resonance spectroscopy (1 in 10) as ASb: Peak area of cefmenoxime from the standard solu-
directed under Nuclear Magnetic Resonance Spectroscopy tion (2)
<2.21>, using tetramethylsilane for nuclear magnetic reso- ATa: Peak area of 1-methyl-1H-tetrazol-5-thiol from the
nance spectroscopy as an internal reference compound: it ex- sample solution
hibits two single signals, A and B, at around d 3.9 ppm, and ST: Total area of the peaks other than 1-methyl-1H-
a single signal C at around d 6.8 ppm. The ratio of the inte- tetrazol-5-thiol and other than cefmenoxime from the
grated intensity of each signal, A:B:C, is about 3:3:1. sample solution
(4) Dissolve 10 mg of Cefmenoxime Hydrochloride in 1
mL of diluted sodium carbonate TS (1 in 20), add 5 mL of
acetic acid (100) and 2 drops of silver nitrate TS: a white pre-
cipitate is formed.
the Assay.
D : －27 – －359(1 g, 0.1 mol/L Time span of measurement: About 2.5 times as long as the
phosphate buffer solution (pH 6.8), 100 mL, 100 mm). retention time of cefmenoxime.
0.10 g of Cefmenoxime Hydrochloride in 150 mL of water is
Purity (1) Clarity and color of solution—A solution ob- the standard solution (1), add the mobile phase to make
tained by dissolving 1.0 g of Cefmenoxime Hydrochloride in exactly 100 mL. Confirm that the peak area of 1-methyl-1H-
10 mL of diluted sodium carbonate TS (1 in 4) is clear and tetrazol-5-thiol obtained from 10 mL of this solution is
colorless to light yellow. equivalent to 4.5 to 5.5z of that obtained from 10 mL of the
(2) Heavy metals <1.07>—Proceed with 1.0 g of Cef- standard solution (1). Then, measure exactly 2 mL of the
menoxime Hydrochloride according to Method 4, and per- standard solution (2), add the mobile phase to make exactly
form the test. Prepare the control solution with 2.0 mL of 100 mL. Confirm that the peak area of cefmenoxime ob-
Standard Lead Solution (not more than 20 ppm). tained from 10 mL of this solution is equivalent to 1.5 to
(3) Arsenic <1.11>—Prepare the test solution with 1.0 g 2.5z of that obtained from 10 mL of the standard solution
of Cefmenoxime Hydrochloride according to Method 4 and (2).
adding 10 mL of dilute hydrochloric acid to the residue after System repeatability: When the test is repeated 6 times
cooling, and perform the test (not more than 2 ppm). with 10 mL of the standard solution (1) under the above
(4) Related substances—Weigh accurately about 0.1 g of operating conditions, the relative standard deviation of the
Cefmenoxime Hydrochloride, dissolve in 20 mL of 0.1 peak area of 1-methyl-1H-tetrazol-5-thiol is not more than
mol/L phosphate buffer solution (pH 6.8) and add the 1.0z.
mobile phase to make exactly 100 mL. Pipet 4 mL of this
determination and methanol for water determination (2:1)).
accurately about 10 mg of 1-methyl-1H-tetrazol-5-thiol, and
dissolve in the mobile phase to make exactly 100 mL. Pipet 4 Assay Weigh accurately an amount of Cefmenoxime Hy-
mL of this solution, add the mobile phase to make exactly drochloride and Cefmenoxime Hydrochloride RS, equiva-
250 mL, and use this solution as the standard solution (1). lent to about 50 mg (potency), dissolve each in 10 mL of 0.1
Weigh accurately about 0.1 g of Cefmenoxime Hydrochlo- mol/L phosphate buffer solution (pH 6.8) and add the
ride RS, dissolve in 20 mL of 0.1 mol/L phosphate buffer mobile phase to make exactly 50 mL. Pipet 4 mL each of
solution (pH 6.8) and add the mobile phase to make exactly these solutions, add exactly 20 mL of the internal standard
100 mL. Pipet 1 mL of this solution, add the mobile phase to solution and the mobile phase to make 50 mL, and use these
make exactly 250 mL, and use this solution as the standard solutions as the sample solution and the standard solution,
solution (2). Perform the test immediately after preparation respectively. Perform the test with 10 mL each of the sample
of these solutions with exactly 10 mL each of the sample solution and standard solution as directed under Liquid
solution and standard solutions (1) and (2) as directed under Chromatography <2.01> according to the following condi-
Liquid Chromatography <2.01> according to the following tions, and calculate the ratios, QT and QS, of the peak area
conditions. Determine each peak area of these solutions by of cefmenoxime to that of the internal standard.
the automatic integration method, and calculate the amounts
Amount [ mg (potency)] of cefmenoxime (C16H17N9O5S3)
of 1-methyl-1H-tetrazol-5-thiol and the total related sub-
＝ MS × QT/QS × 1000
stance by the following formula: the amount of 1-methyl-
1H-tetrazol-5-thiol is not more than 1.0z, and the total MS: Amount [mg (potency)] of Cefmenoxime Hydrochlo-
related substance is not more than 3.0z. ride RS taken
JP XVII Official Monographs / Cefmetazole Sodium 627
Internal standard solution—A solution of phthalimide in pare the spectrum with the Reference Spectrum: both spectra
methanol (3 in 2000). exhibit similar intensities of absorption at the same wave
Operating conditions— numbers.
Detector: An ultraviolet absorption photometer (wave- (3) Determine the 1H spectrum of a solution of Cefmeta-
length: 254 nm). zole Sodium in heavy water for nuclear magnetic resonance
Column: A stainless steel column 4 mm in inside diameter spectroscopy (1 in 10) as directed under Nuclear Magnetic
and 15 cm in length, packed with octadecylsilanized silica gel Resonance Spectroscopy <2.21>, using sodium 3-trimethyl-
for liquid chromatography (5 mm in particle diameter). silylpropanesulfonate for nuclear magnetic resonance spec-
Column temperature: A constant temperature of about troscopy as an internal reference compound: it exhibits
259 C. single signals, A, B and C, at around d 3.6 ppm, at around
Mobile phase: A mixture of water, acetonitrile and acetic d 4.1 ppm and at around d 5.2 ppm, respectively. The ratio
acid (100) (50:10:1). of integrated intensity of each signal, A:B:C, is about 3:3:1.
Flow rate: Adjust so that the retention time of cef- (4) Cefmetazole Sodium responds to the Qualitative
menoxime is about 8 minutes. Tests <1.09> (1) for sodium salt.
D : ＋73 – ＋859(0.25 g, water,
25 mL, 100 mm).
ditions, cefmenoxime and the internal standard are eluted in pH <2.54> Dissolve 1.0 g of Cefmetazole Sodium in 10 mL
this order with the resolution between these peaks being not of water: the pH of the solution is between 4.2 and 6.2.
less than 2.3.
of Cefmetazole Sodium in 10 mL of water: the solution is
clear, and has no more color than the following control
solution.
of the peak areas of cefmenoxime to that of the internal
Control solution: To a mixture of exactly 0.5 mL of
Cobalt (II) Chloride CS and exactly 5 mL of Iron (III)
Containers and storage Containers—Hermetic containers. Chloride CS add water to make exactly 50 mL. To exactly 15
mL of this solution add water to make exactly 20 mL.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Cef-
Cefmetazole Sodium metazole Sodium according to Method 2, and perform the
セフメタゾールナトリウム Lead Solution (not more than 20 ppm).
of Cefmetazole Sodium according to Method 3, and perform
(4) Related substances—Dissolve 0.50 g of Cefmetazole
Sodium in 10 mL of water, and use this solution as the sam-
ple solution. Pipet 4 mL, 2 mL, 1 mL, 0.5 mL and 0.25 mL
of the sample solution, add water to them to make exactly
100 mL, and use these solutions as the standard solutions
C15H16N7NaO5S3: 493.52
(1), (2), (3), (4) and (5), respectively. Separately, dissolve
Monosodium (6R,7R)-7-
0.10 g of 1-methyl-1H-tetrazole-5-thiol in water to make ex-
{[(cyanomethylsulfanyl)acetyl]amino}-7-methoxy-3-
actly 100 mL, and use this solution as the standard solution
(1-methyl-1H-tetrazol-5-ylsulfanylmethyl)-8-oxo-5-thia-
(6). Perform the test with these solutions as directed under
1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate
[56796-20-4]
sample solution and standard solutions (1) to (6) on a plate
of silica gel for thin-layer chromatography. Develop with a
Cefmetazole Sodium contains not less than 860 mg
mixture of 1-butanol, water and acetic acid (100) (4:1:1) to a
distance of about 12 cm, and air-dry the plate. Allow the
plate to stand in iodine vapor: the spot obtained from the
Cefmetazole Sodium is expressed as mass (potency) of
sample solution corresponding to the spot obtained from the
cefmetazole (C15H17N7O5S3: 471.53).
standard solution (6) is not more intense than the spot ob-
Description Cefmetazole Sodium occurs as a white to light tained from the standard solution (6), and the spots other
yellowish white, powder or mass. than this spot and other than the principal spot are not more
It is very soluble in water, freely soluble in methanol, intense than the spot from the standard solution (1). Fur-
slightly soluble in ethanol (95), and very slightly soluble in thermore, the total amount of the spots other than the prin-
tetrahydrofuran. cipal spot from the sample solution, calculated by the com-
It is hygroscopic. parison with the spots from the standard solutions (1), (2),
(3), (4) and (5), is not more than 8.0z.
solution of Cefmetazole Sodium (1 in 40,000) as directed Water <2.48> Not more than 1.0z (1 g, volumetric titra-
under Ultraviolet-visible Spectrophotometry <2.24>, and tion, direct titration).
Assay Weigh accurately an amount of Cefmetazole So-
dium and Cefmetazole RS, equivalent to about 50 mg (po-
wavelengths.
tency), and dissolve each in the mobile phase to make exactly
25 mL. Pipet 1 mL each of these solutions, add exactly 10
metazole Sodium as directed in the potassium bromide disk
mL of the internal standard solution, and use these solutions
628 Cefmetazole Sodium for Injection / Official Monographs JP XVII
as the sample solution and the standard solution, respec- <2.25>, and compare the spectrum with the Reference Spec-
tively. Perform the test with 10 mL each of the sample solu- trum: both spectra exhibit similar intensities of absorption at
tion and standard solution as directed under Liquid Chroma- the same wave numbers.
pH <2.54> Take an amount of Cefmetazole Sodium for In-
calculate the ratios, QT and QS, of the peak area of cefmeta-
jection equivalent to 1.0 g (potency) of Cefmetazole Sodium,
zole to that of the internal standard.
and dissolve in 10 mL of water: the pH of the solution is 4.2
Amount [mg (potency)] of cefmetazole (C15H17N7O5S3) to 6.2.
＝ MS × QT/QS × 1000
MS: Amount [mg (potency)] of Cefmetazole RS taken amount of Cefmetazole Sodium for Injection, equivalent to
1.0 g (potency) of Cefmetazole Sodium, in 10 mL of water:
Internal standard solution—A solution of methyl para-
the solution is clear and the color is not darker than the fol-
lowing control solution.
Control solution: Pipet 5 mL of Iron (III) Chloride CS
and 0.5 mL of Cobalt (II) Chloride CS, and add water to
length: 214 nm).
make exactly 50 mL. Pipet 15 mL of this solution, and add
water to make exactly 20 mL.
(2) Related substances—Proceed as directed in the Purity
(4) under Cefmetazole Sodium.
259 C. Bacterial endotoxins <4.01> Less than 0.06 EU/mg (po-
Mobile phase: Dissolve 5.75 g of ammonium dihydrogen- tency).
phosphate in 700 mL of water, add 280 mL of methanol, 20
mL of tetrahydrofuran and 3.2 mL of 40z tetrabutylammo-
nium hydroxide TS, and adjust to pH 4.5 with phosphoric
acid. Foreign particulate matter <6.06> Perform the test accord-
Flow rate: Adjust so that the retention time of cefmetazole ing to Method 2: it meets the requirement.
is about 8 minutes.
ment.
mL of the standard solution under the above operating con- Sterility <4.06> Perform the test according to the Mem-
ditions, cefmetazole and the internal standard are eluted in brane filtration method: it meets the requirement.
Assay Take 10 containers of Cefmetazole Sodium for In-
less than 10.
jection, dissolve the contents of each in the mobile phase,
rinse each of the containers with the mobile phase, combine
the rinse with the respective previous solution, and add the
mobile phase to make exactly 500 mL. Take exactly a
of the peak area of cefmetazole to that of the internal
volume of this solution equivalent to about 0.2 g (potency)
of Cefmetazole Sodium, and add the mobile phase to make
Containers and storage Containers—Hermetic containers. exactly 100 mL. Pipet 1 mL of this solution, add exactly 10
mL of the internal standard solution, and use this solution as
the sample solution. Separately, weigh accurately an amount
Cefmetazole Sodium for Injection of Cefmetazole RS, equivalent to about 50 mg (potency),
and dissolve in the mobile phase to make exactly 25 mL.
注射用セフメタゾールナトリウム Pipet 1 mL of this solution, add exactly 10 mL of the inter-
nal standard solution, and use this solution as the standard
solution. Then, proceed as directed in the Assay under
Cefmetazole Sodium for Injection is a preparation
Cefmetazole Sodium.
for injection which is dissolved before use.
It contains not less than 90.0z and not more than Amount [mg (potency)] of cefmetazole (C15H17N7O5S3)
110.0z of the labeled potency of cefmetazole ＝ M S × QT / QS × 4
(C15H17N7O5S3: 471.53).
MS: Amount [mg (potency)] of Cefmetazole RS taken
Internal standard solution—A solution of methyl parahy-
tions, with Cefmetazole Sodium.
droxybenzoate in the mobile phase (1 in 10,000).
Description Cefmetazole Sodium for Injection is a white to
light yellow powder or masses.
It is hygroscopic.
solution of Cefmetazole Sodium for Injection (1 in 40,000)
metazole Sodium for Injection as directed in the potassium
JP XVII Official Monographs / Cefodizime Sodium 629
of Cefminox Sodium Hydrate according to Method 3, and

Cefminox Sodium Hydrate perform the test (not more than 1 ppm).
Water <2.48> Not less than 18.0z and not more than
セフミノクスナトリウム水和物
20.0z (0.1 g, volumetric titration, direct titration).
(i) Test organism—Escherichia coli NIHJ
(ii) Culture medium—Use the medium iii in 3) under (1)
Agar media for seed and base layer. Adjust the pH of the
medium so that it will be 6.5 to 6.6 after sterilization.
C16H20N7NaO7S3.7H2O: 667.66
(iii) Standard solution—Weigh accurately an amount of
Monosodium (6R,7S )-7-{2-[(2S )-2-amino-2-
Cefminox Sodium RS, equivalent to about 40 mg (potency),
carboxyethylsulfanyl]acetylamino}-7-methoxy-3-(1-methyl-
1H-tetrazol-5-ylsulfanylmethyl)-8-oxo-5-thia-1-
azabicyclo[4.2.0]oct-2-ene-2-carboxylate heptahydrate
stock solution. Keep the standard stock solution at 59 C or
[75498-96-3]
below and use within 7 days. Take exactly a suitable amount
of the standard stock solution before use, add 0.05 mol/L
Cefminox Sodium Hydrate contains not less than
phosphate buffer solution (pH 7.0) to make solutions so that
each mL contains 40 mg (potency) and 20 mg (potency), and
per mg, calculated on the anhydrous basis. The po-
use these solutions as the high concentration standard solu-
tency of Cefminox Sodium Hydrate is expressed as
tion and the low concentration standard solution, respec-
mass (potency) of cefminox (C16H21N7O7S3: 519.58).
tively.
Description Cefminox Sodium Hydrate occurs as a white (iv) Sample solution—Weigh accurately an amount of
to light yellow crystalline powder. Cefminox Sodium Hydrate equivalent to about 40 mg (po-
It is freely soluble in water, sparingly soluble in methanol, tency), dissolve in 0.05 mol/L phosphate buffer solution (pH
and practically insoluble in ethanol (99.5). 7.0) to make exactly 50 mL. Take exactly a suitable amount
of this solution, add 0.05 mol/L phosphate buffer solution
(pH 7.0) to make solutions so that each mL contains 40 mg
solution of Cefminox Sodium Hydrate (1 in 50,000) as di-
(potency) and 20 mg (potency), and use these solutions as the
high concentration sample solution and the low concentra-
tion sample solution, respectively.
the spectrum of a solution of Cefminox Sodium RS prepared
(v) Procedure—Incubate between 329C and 359C.
in the same manner as the sample solution: both spectra
exhibit similar intensities of absorption at the same wave- Containers and storage Containers—Hermetic containers.
lengths.
minox Sodium Hydrate as directed in the potassium bromide Cefodizime Sodium
compare the spectrum with the Reference Spectrum or the セフォジジムナトリウム
spectrum of Cefminox Sodium RS: both spectra exhibit
(3) Determine the 1H spectrum of a solution of Cefmi-
nox Sodium Hydrate in heavy water for nuclear magnetic
trimethylsilylpropanesulfonate for nuclear magnetic reso-
nance spectroscopy as an internal reference compound: it
exhibits a multiple signal, A, at around d 3.2 ppm, a single C20H18N6Na2O7S4: 628.63
signal, B, at around d 3.5 ppm, a single signal, C, at around Disodium (6R,7R)-7-[(Z )-2-(2-aminothiazol-4-yl)-
d 4.0 ppm, and a single signal, D, at around d 5.1 ppm. The 2-(methoxyimino)acetylamino]-3-[(5-carboxylatomethyl-
ratio of integrated intensity of each signal, A:B:C:D, is 4-methylthiazol-2-yl)sulfanylmethyl]-8-oxo-5-thia-1-
about 2:3:3:1. azabicyclo[4.2.0]oct-2-ene-2-carboxylate
(4) Cefminox Sodium Hydrate responds to the Qualita- [86329-79-5]
Cefodizime Sodium contains not less than 890 mg
D : ＋62 – ＋729(50 mg, water,
(potency) per mg, calculated on the anhydrous and
10 mL, 100 mm).
ethanol-free basis. The potency of Cefodizime
pH <2.54> Dissolve 0.70 g of Cefminox Sodium Hydrate in Sodium is expressed as mass (potency) of cefodizime
10 mL of water: the pH of the solution is between 4.5 and (C20H20N6O7S4: 584.67).
6.0.
Description Cefodizime Sodium occurs as a white to light
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of yellowish white crystalline powder.
Cefminox Sodium Hydrate according to Method 2, and per- It is very soluble in water, and practically insoluble in
form the test. Prepare the control solution with 2.0 mL of acetonitrile and in ethanol (99.5).
630 Cefodizime Sodium / Official Monographs JP XVII
solution of Cefodizime Sodium (1 in 50,000) as directed System suitability—
under Ultraviolet-visible Spectrophotometry <2.24>, and System performance, and system repeatability: Proceed as
compare the spectrum with the Reference Spectrum or the directed in the system suitability in the Assay.
spectrum of a solution of Cefodizime Sodium RS prepared Test for required detectability: Measure exactly 2 mL of
in the same manner as the sample solution: both spectra the standard solution, and add the mobile phase to make ex-
exhibit similar intensities of absorption at the same wave- actly 20 mL. Confirm that the peak area of cefodizime ob-
lengths. tained from 5 mL of this solution is equivalent to 7 to 13z of
(2) Determine the infrared absorption spectrum of that obtained from 5 mL of the standard solution.
Cefodizime Sodium as directed in the potassium bromide (5) Ethanol—Weigh accurately about 1 g of Cefodizime
disk method under Infrared Spectrophotometry <2.25>, and Sodium, and dissolve in water to make exactly 10 mL. Pipet
compare the spectrum with the Reference Spectrum or the 2 mL of this solution, add exactly 2 mL of the internal stand-
spectrum of Cefodizime Sodium RS: both spectra exhibit ard solution, and use this solution as the sample solution.
similar intensities of absorption at the same wave numbers. Separately, weigh accurately about 2 g of ethanol for gas
(3) Determine the 1H spectrum of a solution of Cefodi- chromatography, and add water to make exactly 1000 mL.
zime Sodium in heavy water for nuclear magnetic resonance Pipet 2 mL of this solution, add exactly 2 mL of the internal
spectroscopy (1 in 10) as directed under Nuclear Magnetic standard solution, and use this solution as the standard solu-
Resonance Spectroscopy <2.21>, using sodium 3-trimethyl- tion. Perform the test with 10 mL each of the sample solution
silylpropanesulfonate for nuclear magnetic resonance spec- and standard solution as directed under Gas Chromatogra-
troscopy as an internal reference compound: it exhibits phy <2.02> according to the following conditions, and calcu-
single signals, A, B and C, at around d 2.3 ppm, at around d late the ratios, QT and QS, of the peak area of ethanol to that
4.0 ppm, and at around d 7.0 ppm. The ratio of the inte- of the internal standard: the amount of ethanol is not more
grated intensity of these signals, A:B:C, is about 3:3:1. than 2.0z.
(4) Cefodizime Sodium responds to the Qualitative Tests
Amount (z) of ethanol ＝ MS/MT × QT/QS
<1.09> (1) for sodium salt.
MS: Amount (g) of ethanol for gas chromatography taken
D : －56 – －629(0.2 g calculated
MT: Amount (g) of Cefodizime Sodium taken
on the anhydrous and ethanol-free basis, water, 20 mL, 100
mm). Internal standard solution—A solution of 1-propanol (1 in
400).
pH <2.54> Dissolve 1.0 g of Cefodizime Sodium in 10 mL
Purity (1) Clarity and color of solution—Dissolve 1.0 g Column: A glass column 3.2 mm in inside diameter and
of Cefodizime Sodium in 10 mL of water: the solution is 3 m in length, packed with tetrafluoroethylene polymer for
clear and pale yellow to light yellow. gas chromatography (180 – 250 mm in particle diameter)
(2) Heavy metals <1.07>—Weigh 1.0 g of Cefodizime coated in 15z with polyethylene glycol 20 M.
Sodium in a crucible, cover loosely, and carbonize by gentle Column temperature: A constant temperature of about
heating. After cooling, add 2 mL of sulfuric acid, heat 1009C.
gradually until the white fumes are no longer evolved, and Carrier gas: Nitrogen.
ignite between 5009C and 6009C. Proceed according to Flow rate: Adjust so that the retention time of ethanol is
Method 2, and perform the test. Prepare the control solution about 3 minutes.
with 2.0 mL of Standard Lead Solution (not more than 20 System suitability—
ppm). System performance: When the procedure is run with 10
(3) Arsenic <1.11>—Prepare the test solution with 1.0 g mL of the standard solution under the above operating con-
of Cefodizime Sodium according to Method 3, and perform ditions, ethanol and the internal standard are eluted in this
the test (not more than 2 ppm). order with the resolution between these peaks being not less
(4) Related substances—Dissolve 30 mg of Cefodizime than 2.5.
Sodium in 10 mL of the mobile phase, and use this solution System repeatability: When the test is repeated 6 times
as the sample solution. Pipet 1 mL of the sample solution, with 10 mL of the standard solution under the above operat-
add the mobile phase to make exactly 100 mL, and use this ing conditions, the relative standard deviation of the ratios
solution as the standard solution. Perform the test with ex- of the peak area of ethanol to that of the internal standard is
actly 5 mL each of the sample solution and standard solution not more than 2.0z.
to the following conditions, and determine each peak area by
the automatic integration method: the area of the peaks
other than cefodizime from the sample solution is not larger Assay Weigh accurately an amount of Cefodizime Sodium
than the peak area of cefodizime from the standard solution, and Cefodizime Sodium RS, equivalent to about 50 mg (po-
and the total area of the peaks other than cefodizime from tency), add exactly 10 mL of the internal standard solution
the sample solution is not larger than 3 times the peak area to dissolve, add water to make 100 mL, and use these solu-
of cefodizime from the standard solution. tions as the sample solution and standard solution. Perform
Operating conditions— the test with 10 mL each of the sample solution and standard
Detector, column, column temperature, mobile phase, and solution as directed under Liquid Chromatography <2.01>
flow rate: Proceed as directed in the operating conditions in according to the following conditions, and calculate the
the Assay. ratios, QT and QS, of the peak area of cefodizime to that of
Time span of measurement: About 4 times as long as the the internal standard.
retention time of cefodizime, beginning after the solvent
Amount [ mg (potency)] of cefodizime (C20H20N6O7S4)
peak.
＝ MS × QT/QS × 1000
JP XVII Official Monographs / Cefoperazone Sodium 631
MS: Amount [mg (potency)] of Cefodizime Sodium RS wavelengths.

taken (2) Determine the 1H spectrum of a solution of Cefoper-
azone Sodium in heavy water for nuclear magnetic resonance
Internal standard solution—A solution of anhydrous
spectroscopy (1 in 10) as directed under Nuclear Magnetic
caffeine (3 in 400).
Resonance Spectroscopy <2.21>, using sodium 3-trimethyl-
silylpropanesulfonate for nuclear magnetic resonance spec-
troscopy as an internal reference compound: it exhibits a
length: 254 nm).
triplet signal A at around d 1.2 ppm, and a pair of double
signals, B and C, at around d 6.8 ppm and at around d 7.3
ppm. The ratio of integrated intensity of these signals,
A:B:C, is about 3:2:2.
(3) Cefoperazone Sodium responds to the Qualitative
259 C.
phosphate and 0.20 g of anhydrous disodium hydrogen Optical rotation <2.49> [a]20
D : －15 – －259(1 g, water, 100
phosphate in a suitable amount of water, and add 80 mL of mL, 100 mm).
acetonitrile and water to make 1000 mL.
pH <2.54> Dissolve 1.0 g of Cefoperazone Sodium in 4 mL
Flow rate: Adjust so that the retention time of cefodizime
is about 5 minutes.
System suitability— Purity (1) Clarity and color of solution—Dissolve 1.0 g
System performance: When the procedure is run with 10 of Cefoperazone Sodium in 10 mL of water: the solution is
mL of the standard solution under the above operating con- clear, and its absorbance at 400 nm, determined as directed
ditions, cefodizime and the internal standard are eluted in under Ultraviolet-visible Spectrophotometry <2.24>, is not
this order with the resolution between these peaks being not more than 0.18.
less than 6. (2) Heavy metals <1.07>—Proceed with 1.0 g of Cefoper-
System repeatability: When the test is repeated 6 times azone Sodium according to Method 4, and perform the test.
with 10 mL of the standard solution under the above operat- Prepare the control solution with 2.0 mL of Standard Lead
ing conditions, the relative standard deviation of the ratios Solution (not more than 20 ppm).
of the peak area of cefodizime to that of the internal stand- (3) Arsenic <1.11>—Prepare the test solution with 1.0 g
ard is not more than 2.0z. of Cefoperazone Sodium according to Method 4, and per-
(4) Related substances—Dissolve 0.1 g of Cefoperazone
Sodium in 100 mL of water, and use this solution as the sam-
ple solution. Pipet 1 mL of the sample solution, add water to
Cefoperazone Sodium make exactly 50 mL, and use this solution as the standard
セフォペラゾンナトリウム
ditions, and determine the areas of each peak by the auto-
matic integration method. Calculate the percentages of each
peak area from the sample solution to 50 times of the peak
area of cefoperazone from the standard solution: the related
substance I with the retention time of about 8 minutes is not
more than 5.0z, the related substance II with that of about
17 minutes is not more than 1.5z, and the total of all related
C25H26N9NaO8S2: 667.65
substances is not more than 7.0z. For the peak areas of the
Monosodium (6R,7R)-7-{(2R)-2-[(4-ethyl-2,3-
related substances I and II, multiply their relative response
dioxopiperazine-1-carbonyl)amino]-2-(4-
factors, 0.90 and 0.75, respectively.
hydroxyphenyl)acetylamino}-3-(1-methyl-1H-tetrazol-
5-ylsulfanylmethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-
2-ene-2-carboxylate
[62893-20-3]
the Assay.
Cefoperazone Sodium contains not less than 871 mg
retention time of cefoperazone, beginning after the solvent
peak.
Cefoperazone Sodium is expressed as mass (potency)
of cefoperazone (C25H27N9O8S2: 645.67).
Description Cefoperazone Sodium occurs as a white to yel- Confirm that the peak area of cefoperazone obtained from
lowish white crystalline powder. 25 mL of this solution is equivalent to 3.5 to 6.5z of that ob-
It is very soluble in water, soluble in methanol, and tained from 25 mL of the standard solution.
slightly soluble in ethanol (99.5). System performance: When the procedure is run with 25
solution of Cefoperazone Sodium (1 in 50,000) as directed
factor of the peak of cefoperazone are not less than 5000 and
632 Cefoperazone Sodium and Sulbactam Sodium for Injection / Official Monographs JP XVII
ing conditions, the relative standard deviation of the peak Cefoperazone Sodium and Sulbac-
areas of cefoperazone is not more than 2.0z.
tam Sodium for Injection
tion, direct titration). 注射用セフォペラゾンナトリウム･スルバクタムナトリウ
ム
Assay Weigh accurately an amount of Cefoperazone So-
dium equivalent to about 0.1 g (potency), and dissolve in
water to make exactly 100 mL. Pipet 5 mL of this solution, Cefoperazone Sodium and Sulbactam Sodium for
add exactly 5 mL of the internal standard solution, and use Injection is a preparation for injection which is dis-
this solution as the sample solution. Separately, weigh accu- solved before use.
rately an amount of Cefoperazone RS equivalent to about It contains not less than 90.0z and not more
20 mg (potency), dissolve in 1 mL of 0.1 mol/L phosphate than 110.0z of the labeled potency of cefoperazone
buffer solution (pH 7.0) and add water to make exactly 20 (C25H27N9O8S2: 645.67), and not less than 95.0z and
mL. Pipet 5 mL of this solution, add exactly 5 mL of the in- not more than 110.0z of the labeled potency of sul-
ternal standard solution, and use this solution as the stand- bactam (C8H11NO5S: 233.24).
Method of Preparation Prepare as directed under Injec-
tions, with Cefoperazone Sodium and Sulbactam Sodium.
tions, and calculate the ratios, QT and QS, of the peak area Description Cefoperazone Sodium and Sulbactam Sodium
of cefoperazone to that of the internal standard. for Injection occurs as white to pale yellowish white, masses
or powder.
Amount [mg (potency)] of cefoperazone (C25H27N9O8S2)
＝ MS × QT/QS × 5000 Identification (1) The retention times of cefoperazone in
the chromatogram obtained from the sample solution and
MS: Amount [mg (potency)] of Cefoperazone RS taken
the standard solution in the Assay are the same, and the
Internal standard solution—A solution of acetanilide in a peak area of cefoperazone obtained from the sample solu-
mixture of water and acetonitrile (43:7) (3 in 8000). tion in the Assay is 0.8 to 1.1 times the peak area of cefoper-
Operating conditions— azone obtained by the test performed with 10 mL of the sam-
Detector: An ultraviolet absorption photometer (wave- ple solution obtained in the Assay as directed under Liquid
length: 254 nm). Chromatography <2.01> according to the following condi-
Column: A stainless steel column 4.6 mm in inside diame- tions.
ter and 15 cm in length, packed with octadecylsilanized silica Operating conditions—
gel for liquid chromatography (5 mm in particle diameter). Column, column temperature, mobile phase, and flow
Column temperature: A constant temperature of about rate: Proceed as directed in the operating conditions in the
359 C. Assay.
Mobile phase: To 57 mL of acetic acid (100) add 139 mL Detector: An ultraviolet absorption photometer (wave-
of triethylamine and water to make 1000 mL. To 20 mL of length: 230 nm).
this solution add 835 mL of water, 140 mL of acetonitrile System suitability—
and 5 mL of dilute acetic acid. System performance: Proceed as directed in the system
Flow rate: Adjust so that the retention time of cefopera- suitability in the Assay.
zone is about 10 minutes. (2) The retention times of sulbactam in the chromato-
System suitability— gram obtained from the sample solution and the standard
System performance: When the procedure is run with 10 solution in the Assay are the same, and the peak area of sul-
mL of the standard solution under the above operating con- bactam obtained from the sample solution in the Assay is 1.4
ditions, the internal standard and cefoperazone are eluted in to 1.9 times the peak area of sulbactam obtained by the test
this order with the resolution between these peaks being not performed with 10 mL of the sample solution obtained in the
less than 5. Assay as directed under Liquid Chromatography <2.01> ac-
System repeatability: When the test is repeated 6 times cording to the following conditions.
with 10 mL of the standard solution under the above operat- Operating conditions—
ing conditions, the relative standard deviation of the ratios Column, column temperature, mobile phase, and flow
of the peak area of cefoperazone to that of the internal rate: Proceed as directed in the operating conditions in the
standard is not more than 1.0z. Assay.
length: 230 nm).
amount of Cefoperazone Sodium and Sulbactam Sodium for
Injection, equivalent to 1.0 g (potency) of Cefoperazone So-
dium, in 20 mL of water is 4.5 to 6.5.
Purity (1) Clarity and color of solution—A solution of an
amount of Cefoperazone Sodium and Sulbactam Sodium for
Injection, equivalent to 0.5 g (potency) of Cefoperazone So-
dium, in 10 mL of water is clear. Perform the test with this
JP XVII Official Monographs / Cefoperazone Sodium and Sulbactam Sodium for Injection 633
solution as directed under Ultraviolet Spectrophotometry Uniformity of dosage units <6.02> It meets the requirement
<2.24>: the absorbance at 425 nm is not more than 0.10. of the Mass variation test (T: 105.0z).
(2) Related substances—Weigh accurately an amount of
Cefoperazone Sodium and Sulbactam Sodium for Injection,
equivalent to 0.1 g (potency) of Cefoperazone Sodium, dis-
solve in the mobile phase to make exactly 50 mL, and use Insoluble particulate matter <6.07> It meets the require-
this solution as the sample solution. Pipet 2 mL of the sam- ment.
and use this solution as the standard solution (1). Weigh ac-
curately about 40 mg of sulbactam sodium for sulbactam
penicillamine, dissolve in 2 mL of water, add 0.5 mL of so- Assay Weigh accurately the mass of the content of not less
dium hydroxide TS, allow to stand at room temperature for than 5 Cefoperazone Sodium and Sulbactam Sodium for In-
10 minutes, then add 0.5 mL of 1 mol/L hydrochloric acid jection. Weigh accurately a portion of the content, equiva-
TS, and add the mobile phase to make exactly 100 mL. Pipet lent to about 50 mg (potency) of Cefoperazone Sodium, dis-
5 mL of this solution, add the mobile phase to make exactly solve in suitable amount of the mobile phase, add exactly 5
50 mL, and use this solution as the standard solution (2). mL of the internal standard solution, add the mobile phase
Perform the test with exactly 10 mL each of the sample solu- to make 50 mL, and use this solution as the sample solution.
tion and the standard solutions (1) and (2) as directed under Separately, weigh accurately about 50 mg (potency) each of
Liquid Chromatography <2.01> according to the following Sulbactam RS and Cefoperazone RS, dissolve in suitable
conditions, and determine each peak area by the automatic amount of the mobile phase, add exactly 5 mL of the inter-
integration method: the area of the peak, having a relative nal standard solution, add the mobile phase to make 50 mL,
retention time of about 0.3 (related substance I) to cefopera- and use this solution as the standard solution. Perform the
zone, obtained from the sample solution is not larger than test with 10 mL each of the sample solution and standard so-
1.75 times the peak area of cefoperazone obtained from the lution as directed under Liquid Chromatography <2.01> ac-
standard solution (1), the area of the peak, having a relative cording to the following conditions, and calculate the ratios,
retention time of about 0.4 (related substance III) and about QTa and QTb of the peak areas of sulbactam and cefopera-
1.3 (related substance II) to cefoperazone, obtained from the zone to that of the internal standard obtained from the sam-
sample solution is not larger than 1/2 times the peak area of ple solution, and the ratios, QSa and QSb of the peak areas of
cefoperazone obtained from the standard solution (1). When sulbactam and cefoperazone to that of the internal standard
determine the peak areas, AT and AS, of sulbactam penicilla- obtained from the standard solution.
mine with the sample solution and the standard solution (2),
Amount [mg (potency)] of sulbactam (C8H11NO5S)
and calculate the amount of sulbactam penicillamine by the
＝ MS1 × QTa/QSa
following equation, it is not more than 1.0z. For the area
of the peak of related substance III, multiply the relative Amount [mg (potency)] of cefoperazone (C25H27N9O8S2)
response factor 0.4. ＝ MS2 × QTb/QSb
Amount of sulbactam penicillamine (z) MS1: Amount [mg (potency)] of Sulbactam RS taken
＝ MS/MT × AT/AS × 5 MS2: Amount [mg (potency)] of Cefoperazone RS taken
MS: Amount (mg) of sulbactam sodium for sulbactam Internal standard solution—A solution of ethyl parahy-
penicillamine taken droxybenzoate (7 in 1000).
MT: Amount (mg) of Cefoperazone Sodium and Sulbac- Operating conditions—
tam Sodium for Injection taken Detector: An ultraviolet absorption photometer (wave-
length: 220 nm).
Column, column temperature, mobile phase, and flow
rate: Proceed as directed in the operating conditions in the
Assay.
359C.
length: 230 nm).
Mobile phase: A mixture of 0.005 mol/L tetrabutylammo-
nium hydroxide TS and acetonitrile for liquid chromatogra-
System performance: To 1 mL of the standard solution (1)
phy (3:1).
add 1 mL of the standard solution (2). When the procedure
Flow rate: Adjust so that the retention time of sulbactam
is run with 10 mL of this solution under the above operating
is about 7 minutes.
conditions, sulbactam penicillamine, sulbactam and cefoper-
azone are eluted in this order with the resolutions between
the peaks, sulbactam penicillamine and sulbactam, and sul-
bactam and cefoperazone, being not less than 4 and not less
ditions, sulbactam, the internal standard, and cefoperazone
than 5, respectively.
peaks being not less than 1.5.
with 10 mL of the standard solution (2) under the above
peak area of sulbactam penicillamine is not more than 2.0z.
ing conditions, the relative standard deviation of the peak of
Water <2.48> Not more than 1.0z (1 g, volumetric titra- sulbactam is not more than 1.0z.
Bacterial endotoxins <4.01> Less than 0.060 EU/mg (po- Plastic containers for aqueous injections may be used.
tency).
634 Cefotaxime Sodium / Official Monographs JP XVII
Cefotaxime Sodium follows: To 1.0 mL of 0.005 mol/L sulfuric acid VS add 1
mL of dilute hydrochloric acid and water to make 50 mL
セフォタキシムナトリウム (not more than 0.048z).
Cefotaxime Sodium according to Method 2, and perform the
of Cefotaxime Sodium according to Method 3, and perform
C16H16N5NaO7S2: 477.45
(5) Related substances—Perform the test with 10 mL of
Monosodium (6R,7R)-3-acetoxymethyl-7-[(Z)-2-(2-
the sample solution obtained in the Assay as directed under
aminothiazol-4-yl)-2-(methoxyimino)acetylamino]-8-oxo-5-
thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate
conditions, and determine each peak area obtained from the
[64485-93-4]
chromatogram by the automatic integration method, and
calculated the amounts of them by the area percentage
Cefotaxime Sodium contains not less than 916 mg
method: the amount of the peak other than cefotaxime is not
more than 1.0z and the total amount of these peaks is not
calculated on the dried basis. The potency of
more than 3.0z.
Cefotaxime Sodium is expressed as mass (potency) of
cefotaxime (C16H17N5O7S2: 455.47).
Detector, column, column temperature, mobile phase A,
Description Cefotaxime Sodium occurs as white to light mobile phase B, flowing of mobile phase, and flow rate:
yellowish white crystalline powder. Proceed as directed in the operating conditions in the Assay.
It is freely soluble in water, sparingly soluble in methanol, Time span of measurement: About 3.5 times as long as the
and very slightly soluble in ethanol (95). retention time of cefotaxime, beginning after the solvent
peak.
Identification (1) Dissolve 2 mg of Cefotaxime Sodium in
0.01 mol/L hydrochloric acid TS to make 100 mL. Deter-
mine the absorption spectrum of this solution as directed
the standard solution, and add the mobile phase A to make
exactly 100 mL. Pipet 2 mL of this solution, and add the
wavelengths.
mobile phase A to make exactly 20 mL. Confirm that the
peak area of cefotaxime obtained from 10 mL of this solution
Cefotaxime Sodium as directed in the potassium bromide
is equivalent to 0.15 to 0.25z of that obtained from 10 mL
of the standard solution.
spectra exhibit similar intensities of absorption at the same Loss on drying <2.41> Not more than 3.0z (1 g, 1059C,
wave numbers. 3 hours).
Assay Weigh accurately an amount of Cefotaxime Sodium
Cefotaxime Sodium in heavy water for nuclear magnetic
and Cefotaxime RS, equivalent to about 40 mg (potency),
dissolve each in the mobile phase A to make exactly 50 mL,
and use these solutions as the sample solution and standard
nance spectroscopy as an internal reference compound: it ex-
hibits three single signals, A, B and C, at around d 2.1 ppm,
at around d 4.0 ppm and at around d 7.0 ppm. The ratio of
the integrated intensity of each signal, A:B:C, is about 3:3:1.
cefotaxime in each solution.
(4) Cefotaxime Sodium responds to the Qualitative Tests
<1.09> (1) for sodium salt. Amount [ mg (potency)] of cefotaxime (C16H17N5O7S2)
＝ MS × AT/AS × 1000
＋58 – ＋649 (0.25 g calcu-
D:
lated on the dried basis, water, 25 mL, 100 mm). MS: Amount [mg (potency)] of Cefotaxime RS taken
pH <2.54> The pH of a solution obtained by dissolving Operating conditions—
1.0 g of Cefotaxime Sodium in 10 mL of water is between Detector: An ultraviolet absorption photometer (wave-
4.5 and 6.5. length: 235 nm).
of Cefotaxime Sodium in 10 mL of water: the solution is
clear, and its absorbance at 430 nm, determined as directed
under Ultraviolet-visible Spectrophotometry <2.24>, is not
309C.
more than 0.40.
Mobile phase A: To 0.05 mol/L disodium hydrogen phos-
(2) Sulfate <1.14>—Dissolve 2.0 g of Cefotaxime Sodium
phate TS add phosphoric acid to adjust the pH to 6.25. To
in 40 mL of water, add 2 mL of dilute hydrochloric acid and
860 mL of this solution add 140 mL of methanol.
water to make 50 mL, shake well, and filter. Discard first 10
Mobile phase B: To 0.05 mol/L disodium hydrogen phos-
mL of the filtrate, and to the subsequent 25 mL of the fil-
phate TS add phosphoric acid to adjust the pH to 6.25. To
trate add water to make 50 mL. Perform the test with this
JP XVII Official Monographs / Cefotetan 635
600 mL of this solution add 400 mL of methanol. with the Reference Spectrum or the spectrum of a solution of
Flowing of mobile phase: Control the gradient by mixing Cefotetan RS prepared in the same manner as the sample so-
the mobile phases A and B as directed in the following table. lution: both spectra exhibit similar intensities of absorption
Time after injection Mobile phase A Mobile phase B (2) Determine the infrared absorption spectrum of
of sample (min) (volz) (volz) Cefotetan as directed in the potassium bromide disk method
0–7 100 0 spectrum with the Reference Spectrum or the spectrum of
7–9 100 → 80 0 → 20 Cefotetan RS: both spectra exhibit similar intensities of ab-
9 – 16 80 20 sorption at the same wave numbers.
16 – 45 80 → 0 20 → 100 (3) Dissolve 50 mg of Cefotetan in 0.5 mL of a solution
45 – 50 0 100 of sodium hydrogen carbonate in heavy water for nuclear
magnetic resonance spectroscopy (1 in 25). Determine the 1H
Flow rate: 1.3 mL per minute (the retention time of spectrum of this solution as directed under Nuclear Magnetic
cefotaxime is about 14 minutes). Resonance Spectroscopy <2.21>, using sodium 3-trimethyl-
System suitability— silylpropanesulfonate for nuclear magnetic resonance spec-
System performance: To 1 mL of the standard solution troscopy as an internal reference compound: it exhibits
add 7.0 mL of water and 2.0 mL of methanol, mix, then add single signals, A, B, C and D, at around d 3.6 ppm, at
25 mg of sodium carbonate decahydrate, and shake. After around d 4.0 ppm, at around d 5.1 ppm and at around d 5.2
allowing to stand for 10 minutes, add 3 drops of acetic acid ppm, respectively. The ratio of the integrated intensity of
(100) and 1 mL of the standard solution, and mix. When the each signal, A:B:C:D, is about 3:3:1:1.
procedure is run with 10 mL of this solution under the above Optical rotation <2.49> [a]20
D : ＋112 – ＋1249(0.5 g calcu-
operating conditions, desacetyl cefotaxime with the relative lated on the anhydrous basis, a solution of sodium hydrogen
retention time being about 0.3 to cefotaxime and cefotaxime carbonate (1 in 200), 50 mL, 100 mm).
peaks being not less than 20, and the symmetry factor of the Purity (1) Clarity and color of solution—Dissolve 1.0 g
peak of cefotaxime is not more than 2. of Cefotetan in 10 mL of a solution of sodium hydrogen car-
System repeatability: When the test is repeated 6 times bonate (1 in 30): the solution is clear, and colorless or light
with 10 mL of the standard solution under the above operat- yellow.
ing conditions, the relative standard deviation of the peak (2) Heavy metals <1.07>—Proceed with 1.0 g of Cefote-
area of cefotaxime is not more than 2.0z. tan according to Method 2, and perform the test. Prepare
Containers and storage Containers—Tight containers. (not more than 20 ppm).
(3) Related substances—Weigh accurately about 0.1 g of
Cefotetan, dissolve in a suitable amount of methanol, add
Cefotetan exactly 2 mL of the internal standard solution and methanol
to make 20 mL, and use this solution as the sample solution.
セフォテタン Separately, weigh accurately about 3 mg of 1-methyl-1H-
tetrazole-5-thiol for liquid chromatography, previously dried
in a desiccator (in vacuum, silica gel) for 2 hours, and about
2 mg of Cefotetan RS, calculated on the anhydrous basis,
dissolve in methanol to make exactly 20 mL. Pipet 2 mL of
this solution, add exactly 2 mL of the internal standard solu-
tion and methanol to make 20 mL, and use this solution as
the standard solution. Perform the test with 5 mL of the sam-
C17H17N7O8S4: 575.62 ple solution and standard solution as directed under Liquid
(6R,7R)-7-{[4-(Carbamoylcarboxymethylidene)-1,3- Chromatography <2.01> according to the following condi-
dithietane-2-carbonyl]amino}-7-methoxy-3-(1-methyl-1H- tions, and calculate the ratios, QTa, QTb, QTc, QTd, QTe and
tetrazol-5-ylsulfanylmethyl)-8-oxo-5-thia-1- QTf, of the peak areas of 1-methyl-1H-tetrazole-5-thiol,
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid cefotetan lactone having the relative retention time of about
[69712-56-7] 0.5 to cefotetan, D2-cefotetan having the relative retention
time of about 1.2, isothiazole substance having the relative
Cefotetan contains not less than 960 mg (potency) retention time of about 1.3, each of other related substances
and not more than 1010 mg (potency) per mg, calcu- and the total of other related substances, to the peak area of
lated on the anhydrous basis. The potency of Cefote- the internal standard, respectively, obtained from the sample
tan is expressed as mass (potency) of cefotetan solution, and the ratios, QSa and QSb, of the peak areas of 1-
(C17H17N7O8S4). methyl-1H-tetrazole-5-thiol and cefotetan, to the peak area
of the internal standard, respectively, obtained from the
Description Cefotetan occurs as white to light yellowish standard solution. Calculate the amount of 1-methyl-1H-
white powder. tetrazole-5-thiol, cefotetan lactone, D2-cefotetan, isothiazole
It is sparingly soluble in methanol, and slightly soluble in substance, each of other related substances and the total of
water and in ethanol (99.5). other related substances from the following equations: the
Identification (1) Determine the absorption spectrum of a amount of 1-methyl-1H-tetrazole-5-thiol is not more than
solution of Cefotetan in phosphate buffer solution for anti- 0.3z, cefotetan lactone is not more than 0.3z, D2-cefotetan
biotics, pH 6.5 (1 in 100,000) as directed under Ultraviolet- is not more than 0.5z, isothiazole substance is not more
visible Spectrophotometry <2.24>, and compare the spectrum than 0.5z, each of other related substances is not more than
0.2z and the total of other related substances is not more
636 Cefotetan / Official Monographs JP XVII
than 0.4z. solution (pH 7.0), water and a solution of tetrabutylammo-
nium hydrogensulfate in acetonitrile (1 in 150) (9:9:2).
1-Methyl-1H-tetrazole-5-thiol (z)
Flow rate: Adjust so that the retention time of l-substance
＝ MSa/MT × QTa/QSa × 1/100
Cefotetan lactone (z) System suitability—
＝ MSb/MT × QTb/QSb × 1/100 System performance: When the procedure is run with 5 mL
of the sample solution under the above operating conditions,
D2-Cefotetan (z)
l-substance and d-substance are eluted in this order with the
＝ MSb/MT × QTc/QSb × 1/100
resolution between these peaks being not less than 1.5.
Isothiazole substance (z) System repeatability: To exactly 1 mL of the sample solu-
＝ MSb/MT × QTd/QSb × 1/100 tion add methanol to make exactly 10 mL. When the test is
repeated 6 times with 5 mL of this solution under the above
Each of other related substances (z)
＝ MSb/MT × QTe/QSb × 1/100
peak area of l-substance is not more than 5.0z.
Total of other related substances (z)
Assay Weigh accurately an amount of Cefotetan and
＝ MSb/MT × QTf/QSb × 1/100
Cefotetan RS, equivalent to about 50 mg (potency), and dis-
MSa: Amount (mg) of 1-methyl-1H-tetrazole-5-thiol taken solve each in phosphate buffer solution for antibiotics, pH
MSb: Amount (mg) of Cefotetan RS, calculated on the an- 6.5 to make exactly 50 mL. Pipet 15 mL each of these solu-
hydrous basis taken tions, add exactly 10 mL of the internal standard solution
MT: Amount (g) of Cefotetan taken and phosphate buffer solution for antibiotics, pH 6.5 to
make 50 mL, and use these solutions as the sample solution
Internal standard solution—A solution of anhydrous
and standard solution. Perform the test with 5 mL each of
caffeine in methanol (3 in 10,000).
area of cefotetan to that of the internal standard.
the Assay.
Time span of measurement: About 3.5 times as long as the Amount [ mg (potency)] of cefotetan (C17H17N7O8S4)
retention time of cefotetan. ＝ MS × QT/QS × 1000
MS: Amount [mg (potency)] of Cefotetan RS taken
suitability in the Assay. Internal standard solution—A solution of anhydrous
Test for required detectability: Measure exactly 15 mL of caffeine (1 in 1000).
the standard solution, and add methanol to make exactly 100 Operating conditions—
mL. Confirm that the peak area of cefotetan obtained from Detector: An ultraviolet absorption photometer (wave-
5 mL of this solution is equivalent to 12 to 18z of that ob- length: 254 nm).
tained from 5 mL of the standard solution. Column: A stainless steel column 4.6 mm in inside diame-
System repeatability: When the test is repeated 6 times ter and 15 cm in length, packed with octadecylsilanized silica
with 5 mL of the standard solution under the above operating gel for liquid chromatography (5 mm in particle diameter).
conditions, the relative standard deviation of the ratio of the Column temperature: A constant temperature of about
peak area of cefotetan to that of the internal standard is not 409C.
more than 2.0z. Mobile phase: Dissolve 11.53 g of phosphoric acid in 1000
mL of water. To 850 mL of this solution add 50 mL of
acetonitrile, 50 mL of acetic acid (100) and 50 mL of metha-
nol.
Residue on ignition <2.44> Not more than 0.1z (1 g). Flow rate: Adjust so that the retention time of cefotetan is
about 17 minutes.
Isomer ratio Dissolve 10 mg of Cefotetan in 20 mL of
methanol, and use this solution as the sample solution. Per-
form the test with 5 mL of the sample solution as directed
tions, the internal standard and cefotetan are eluted in this
automatic integration method. Calculate the amount of the
than 8.
adjacent two peaks appeared at around the retention time of
40 minutes, one having shorter retention time is l-substance
and another having longer retention time is d-substance, by
the area percentage method: the amount of l-substance is not
peak area of cefotetan to that of the internal standard is not
less than 35z and not more than 45z.
more than 1.0z.
Detector: An ultraviolet absorption photometer (wave- Containers and storage Containers—Tight containers.
length: 254 nm). Storage—Light-resistant, and at a temperature not
Column: A stainless steel column 4 mm in inside diameter exceeding 59C.
409 C.
Mobile phase: A mixture of 0.1 mol/L phosphate buffer
JP XVII Official Monographs / Cefotiam Hexetil Hydrochloride 637
and perform the test, using a solution of magnesium nitrate

Cefotiam Hexetil Hydrochloride hexahydrate in ethanol (95) (1 in 5) (not more than 1 ppm).
(3) Related substance 1—Weigh accurately about 50 mg
セフォチアムヘキセチル塩酸塩 of Cefotiam Hexetil Hydrochloride, and dissolve in a mix-
ture of diluted phosphoric acid (1 in 100) and acetonitrile
(4:1) to make exactly 50 mL. Pipet 10 mL of this solution,
add a mixture of diluted phosphoric acid (1 in 100) and
acetonitrile (4:1) to make exactly 25 mL, and use this solu-
about 50 mg of Cefotiam Hexetil Hydrochloride RS, and
dissolve in a mixture of diluted phosphoric acid (1 in 100)
and acetonitrile (4:1) to make exactly 50 mL. Pipet 1 mL of
this solution, add a mixture of diluted phosphoric acid (1 in
100) and acetonitrile (4:1) to make exactly 50 mL, and use
C27H37N9O7S3.2HCl768.76
(1RS )-1-Cyclohexyloxycarbonyloxyethyl (6R,7R)-7-
[2-(2-aminothiazol-4-yl)acetylamino]-3-[1-(2-
dimethylaminoethyl)-1H-tetrazol-5-ylsulfanylmethyl]-
8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate
dihydrochloride
the amount of the related substances by the following equa-
[95789-30-3]
tion: the amount of the related substance having the relative
retention time of about 1.2 to one of the peaks of cefotiam
Cefotiam Hexetil Hydrochloride contains not less
hexetil, which has the larger retention time, is not more than
than 615 mg (potency) and not more than 690 mg (po-
2.0z, and each amount of the other related substances is not
tency) per mg, calculated on the anhydrous basis. The
more than 0.5z. For the peak area, having the relative
potency of Cefotiam Hexetil Hydrochloride is ex-
retention time of about 1.2 to one of the peaks of cefotiam
pressed as mass (potency) of cefotiam (C18H23N9O4S3:
hexetil, which has the larger retention time, multiply the
525.63).
relative response factor, 0.78.
Description Cefotiam Hexetil Hydrochloride occurs as a
white to light yellow powder.
＝ MS/MT × AT/AS × 5
It is very soluble in water, in methanol and in ethanol (95),
freely soluble in dimethylsulfoxide, and slightly soluble in MS: Amount (g) of Cefotiam Hexetil Hydrochloride RS
acetonitrile. taken
It dissolves in 0.1 mol/L hydrochloric acid TS. MT: Amount (g) of Cefotiam Hexetil Hydrochloride taken
It is hygroscopic. AS: Total of two peak areas of cefotiam hexetil from the
standard solution
AT: Each peak area of related substance from the sample
solution of Cefotiam Hexetil Hydrochloride in 0.1 mol/L
solution
hydrochloric acid TS (3 in 125,000) as directed under Ultra-
violet-visible Spectrophotometry <2.24>, and compare the Operating conditions—
spectrum with the Reference Spectrum or the spectrum of a Detector: An ultraviolet absorption photometer (wave-
solution of Cefotiam Hexetil Hydrochloride RS prepared in length: 254 nm).
the same manner as the sample solution: both spectra exhibit Column: A stainless steel column 4 mm in inside diameter
similar intensities of absorption at the same wavelengths. and 15 cm in length, packed with octadecylsilanized silica gel
(2) Determine the 1H spectrum of a solution of Cefotiam for liquid chromatography (5 mm in particle diameter).
Hexetil Hydrochloride in deuterated dimethylsulfoxide for Column temperature: A constant temperature of about
nuclear magnetic resonance spectroscopy (1 in 20) as directed 259C.
under Nuclear Magnetic Resonance Spectroscopy <2.21>, Mobile phase A: A mixture of diluted 0.2 mol/L potas-
using tetramethylsilane for nuclear magnetic resonance spec- sium dihydrogen phosphate TS (1 in 2), acetonitrile and
troscopy as an internal reference compound: it exhibits two acetic acid (100) (72:28:1).
single signals, A and B, at around d 2.8 ppm and at around Mobile phase B: A mixture of acetonitrile, diluted 0.2
d 6.6 ppm, and a multiple signal, C, at around d 6.9 ppm. mol/L potassium dihydrogen phosphate TS (1 in 2) and
The ratio of the integrated intensity of each signal, A:B:C, is acetic acid (100) (60:40:1).
about 6:1:1. Flowing of mobile phase: Adjust so that the mixing rate of
(3) To a solution of Cefotiam Hexetil Hydrochloride the mobile phase A and the mobile phase B is changed lineal-
(1 in 200) add 2 mL of dilute nitric acid and 1 mL of silver ly from 1:0 to 0:1 for 30 minutes.
nitrate TS, and mix: a white precipitate is formed. Flow rate: 0.7 mL per minute.
D : ＋52 – ＋609(0.1 g calculated
the retention time of one of the cefotiam hexetil peaks,
on the anhydrous basis, 0.1 mol/L hydrochloric acid TS, 10
which appears first, beginning after the solvent peak.
mL, 100 mm).
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Test for required detectability: Measure exactly 1 mL of
Cefotiam Hexetil Hydrochloride according to Method 2, and the standard solution, and add a mixture of diluted phos-
perform the test. Prepare the control solution with 2.0 mL of phoric acid (1 in 100) and acetonitrile (4:1) to make exactly
Standard Lead Solution (not more than 10 ppm). 50 mL. Confirm that each area of the two peaks of cefotiam
(2) Arsenic <1.11>—Prepare the test solution with 2.0 g hexetil obtained from 10 mL of this solution is equivalent to
of Cefotiam Hexetil Hydrochloride according to Method 3, 1.6 to 2.4z of that obtained from 10 mL of the standard so-
638 Cefotiam Hexetil Hydrochloride / Official Monographs JP XVII
lution. tions, acetaminophen and cefotiam are eluted in this order
System performance: When the procedure is run with 10 with the resolution between these peaks being not less than 4.
mL of the standard solution under the above operating con- System repeatability: When the test is repeated 6 times
ditions, the resolution between the two peaks of cefotiam with 10 mL of the standard solution under the above operat-
hexetil is not less than 2.0. ing conditions, the relative standard deviation of the peak
System repeatability: When the test is repeated 6 times area of cefotiam is not more than 2.0z.
with 10 mL of the standard solution under the above operat- (5) Total amount of related substances—The total of the
ing conditions, the relative standard deviation of the total of amount of related substances obtained in the Related sub-
the two peak areas of cefotiam hexetil is not more than stance 1 and the Related substance 2 is not more than 6.5z.
2.0z.
(4) Related substance 2—Weigh accurately about 20 mg
of Cefotiam Hexetil Hydrochloride, dissolve in 2 mL of
methanol, add a mixture of a solution of diammonium Residue on ignition <2.44> Not more than 0.1z (1 g).
hydrogen phosphate (79 in 20,000) and acetic acid (100)
Isomer ratio Proceed the test with 20 mL of the sample so-
lution obtained in the Assay as directed under Liquid Chro-
matography <2.01> according to the conditions directed in
of Cefotiam Hydrochloride RS, and dissolve in the mobile
the Assay, and determine the areas of the two peaks, Aa for
the faster peak and Ab for the later peak, closely appeared
add the mobile phase to make exactly 50 mL, and use this so-
each other at the retention time of around 10 minutes: Aa/
(Aa ＋ Ab) is not less than 0.45 and not more than 0.55.
directed under Liquid Chromatography <2.01> according to Assay Weigh accurately an amount of Cefotiam Hexetil
the following conditions, and determine each peak area by Hydrochloride and Cefotiam Hexetil Hydrochloride RS,
the automatic integration method. Calculate the amount of equivalent to about 30 mg (potency), and dissolve each in a
the related substances by the following equation: the mixture of diluted phosphoric acid (1 in 100) and acetonitrile
amounts of the related substances having the relative reten- (4:1) to make exactly 50 mL. Measure exactly 5 mL each of
tion time of about 0.1 and about 0.9 to cefotiam are not these solutions, add exactly 5 mL of the internal standard
more than 1.0z, respectively, and each amount of the solution and a mixture of diluted phosphoric acid (1 in 100)
related substances other than the related substances having and acetonitrile (4:1) to make 50 mL, and use these solutions
the relative retention time of about 0.1 and about 0.9 to as the sample solution and standard solution. Perform the
cefotiam is not more than 0.5z. For the peak area, having test with 20 mL each of the sample solution and standard
the relative retention time of about 0.9 to cefotiam, multiply solution as directed under Liquid Chromatography <2.01>
the relative response factor, 0.76. according to the following conditions, and calculate the
ratios, QT and QS, of the peak area of cefotiam hexetil to
that of the internal standard. For this calculation, the total
＝ M S / M T × AT / AS × 4
of the areas of the two peaks appeared closely each other at
MS: Amount (g) of Cefotiam Hydrochloride RS taken the retention time of around 10 minutes is used as the peak
MT: Amount (g) of Cefotiam Hexetil Hydrochloride taken area of cefotiam hexetil.
AS: Peak area of cefotiam from the standard solution
Amount [ mg (potency)] of cefotiam (C18H23N9O4S3)
AT: Each peak area from the sample solution
＝ MS × QT/QS × 1000
MS: Amount [mg (potency)] of Cefotiam Hexetil Hydro-
chloride RS taken
length: 254 nm).
Column: A stainless steel column 4 mm in inside diameter Internal standard solution—A solution of benzoic acid in a
and 15 cm in length, packed with octadecylsilanized silica gel mixture of diluted phosphoric acid (1 in 100) and acetonitrile
for liquid chromatography (5 mm in particle diameter). (4:1) (7 in 10,000).
Column temperature: A constant temperature of about Operating conditions—
259 C. Detector: An ultraviolet absorption photometer (wave-
Mobile phase: A mixture of a solution of diammonium length: 254 nm).
hydrogen phosphate (79 in 20,000), methanol and acetic acid Column: A stainless steel column 4 mm in inside diameter
(100) (200:10:3). and 15 cm in length, packed with octadecylsilanized silica gel
Flow rate: Adjust so that the retention time of cefotiam is for liquid chromatography (5 mm in particle diameter).
about 15 minutes. Column temperature: A constant temperature of about
Time span of measurement: As long as about 2 times of 259C.
the retention time of cefotiam, beginning after the solvent Mobile phase: A mixture of diluted 0.2 mol/L potassium
peak. dihydrogen phosphate TS (1 in 2), acetonitrile and acetic
System suitability— acid (100) (72:28:1).
Test for required detectability: Measure exactly 1 mL of Flow rate: Adjust so that the retention time of the faster
the standard solution, and add the mobile phase to make peak of cefotiam hexetil is about 9 minutes.
exactly 50 mL. Confirm that the peak area of cefotiam System suitability—
obtained from 10 mL of this solution is equivalent to 1.6 to System performance: When the procedure is run with 20
2.4z of that obtained from 10 mL of the standard solution. mL of the standard solution under the above operating con-
System performance: To 1 mL of a solution of acetamino- ditions, the internal standard and cefotiam hexetil are eluted
phen in the mobile phase (1 in 50,000) add 3 mL of the in this order with the resolution between the two peaks of
standard solution, and mix well. When the procedure is run cefotiam hexetil being not less than 2.0.
with 10 mL of this solution under the above operating condi- System repeatability: When the test is repeated 6 times
JP XVII Official Monographs / Cefotiam Hydrochloride 639
with 20 mL of the standard solution under the above operat- on the anhydrous bases, water, 100 mL, 100 mm).
pH <2.54> Dissolve 1.0 g of Cefotiam Hydrochloride in 10
of the peak area of cefotiam hexetil to that of the internal
of Cefotiam Hydrochloride in 10 mL of water: the solution
is clear, and colorless to yellow.
(2) Heavy metals <1.07>—To 1.0 g of Cefotiam Hydro-
Cefotiam Hydrochloride chloride add 1 mL of sulfuric acid, and heat gently to car-
bonize. After cooling, add 10 mL of a solution of magne-
セフォチアム塩酸塩
sium nitrate hexahydrate in ethanol (95) (1 in 10), fire the
ethanol to burn, then heat gradually to incinerate. If a car-
bonized residue still retains, moisten the residue with a little
amount of sulfuric acid, and ignite again to incinerate. After
cooling, add 2 mL of hydrochloric acid to the residue, heat
on a water bath to dissolve, then heat to dryness. Add 10 mL
of water, and heat to dissolve. After cooling, add ammonia
TS dropwise to adjust to pH 3 – 4, if necessary, filter, wash
the residue on the filter with 10 mL of water, transfer the fil-
trate and washings into a Nessler tube, add water to make 50
C18H23N9O4S3.2HCl: 598.55 mL, and use this solution as the test solution. Prepare the
(6R,7R)-7-[2-(2-Aminothiazol-4-yl)acetylamino]-3- control solution with 2.0 mL of Standard Lead Solution in
[1-(2-dimethylaminoethyl)-1H-tetrazol-5-ylsulfanylmethyl]- the same manner as for preparation of the test solution (not
8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid more than 20 ppm).
dihydrochloride (3) Arsenic <1.11>—Incinerate 1.0 g of Cefotiam Hydro-
[66309-69-1] chloride according to Method 4. After cooling, add 10 mL of
dilute hydrochloric acid to the residue, heat to dissolve on
Cefotiam Hydrochloride contains not less than 810 the water bath, and use this solution as the test solution. Per-
mg (potency) and not more than 890 mg (potency) per form the test (not more than 2 ppm).
Water <2.48> Not more than 7.0z (0.25 g, volumetric
Cefotiam Hydrochloride is expressed as mass (po-
titration, direct titration. Use a mixture of formamide for
tency) of cefotiam (C18H23N9O4S3: 525.63).
water determination and methanol for water determination
Description Cefotiam Hydrochloride occurs as white to (2:1) instead of methanol for water determination).
light yellow, crystals or crystalline powder.
Assay Weigh accurately an amount of Cefotiam Hydro-
It is freely soluble in water, in methanol and in for-
chloride and Cefotiam Hydrochloride RS, equivalent to
mamide, slightly soluble in ethanol (95), and practically in-
about 0.1 g (potency), and dissolve each in the mobile phase
soluble in acetonitrite.
to make exactly 100 mL, and use these solutions as the sam-
Identification (1) Determine the absorption spectrum of a ple solution and the standard solution, respectively. Perform
solution of Cefotiam Hydrochloride (1 in 50,000) as directed the test with exactly 10 mL each of the sample solution and
under Ultraviolet-visible Spectrophotometry <2.24>, and standard solution as directed under Liquid Chromatography
compare the spectrum with the Reference Spectrum or the <2.01> according to the following conditions, and determine
spectrum of a solution of Cefotiam Hydrochloride RS the peak areas, AT and AS, of cefotiam in each solution.
Amount [ mg (potency)] of cefotiam (C18H23N9O4S3)
＝ MS × AT/AS × 1000
wavelengths.
(2) Determine the infrared absorption spectrum of MS: Amount [mg (potency)] of Cefotiam Hydrochloride
Cefotiam Hydrochloride as directed in the potassium chlo- RS taken
the spectrum of Cefotiam Hydrochloride RS: both spectra
length: 254 nm).
numbers.
ter and 125 mm in length, packed with octadecylsilanized
(3) Determine the 1H spectrum of a solution of Cefotiam
Hydrochloride in heavy water for nuclear magnetic reso-
ter).
nance spectroscopy (1 in 10) as directed under Nuclear
259C.
Mobile phase: To 800 mL of 0.05 mol/L disodium
nance spectroscopy as an internal reference compound: it
hydrogenphosphate TS add 0.05 mol/L potassium dihydro-
exhibits single signals, A and B, at around d 3.1 ppm and at
genphosphate TS to adjust the pH to 7.7. To 440 mL of this
around d 6.7 ppm, respectively. The ratio of integrated in-
tensity of each signal, A:B, is about 6:1.
Flow rate: Adjust so that the retention time of cefotiam is
(4) Dissolve 0.1 g of Cefotiam Hydrochloride in 5 mL of
about 14 minutes.
dilute nitric acid, and immediately add 1 mL of silver nitrate
TS: a white precipitate is formed.
System performance: Dissolve 0.04 g of orcine in 10 mL of
D : ＋60 – ＋729(1 g calculated the standard solution. When the procedure is run with 10 mL
640 Cefotiam Hydrochloride for Injection / Official Monographs JP XVII
of the standard solution under the above operating condi- Assay Weigh accurately the contents of not less than 10
tions, orcine and cefotiam are eluted in this order with the Cefotiam Hydrochloride for Injection. Weigh accurately an
resolution between these peaks being not less than 5. amount of the content, equivalent to about 50 mg (potency)
System repeatability: When the test is repeated 6 times of Cefotiam Hydrochloride, dissolve in the mobile phase to
with 10 mL of the standard solution under the above operat- make exactly 50 mL, and use this solution as the sample so-
ing conditions, the relative standard deviation of the peak lution. Separately, weigh accurately about 50 mg (potency)
areas of cefotiam is not more than 1.0z. of Cefotiam Hydrochloride RS, dissolve in the mobile phase
solution. Proceed as directed in the Assay under Cefotiam
Hydrochloride.
Cefotiam Hydrochloride for Amount [ mg (potency)] of cefotiam (C18H23N9O4S3)

＝ MS × AT/AS × 1000
Injection
MS: Amount [mg (potency)] of Cefotiam Hydrochloride
注射用セフォチアム塩酸塩 RS taken
Cefotiam Hydrochloride for Injection is a prepara- Plastic containers for aqueous injections may be used.
tion for injection which is dissolved before use.
than 110.0z of the labeled potency of cefotiam Cefozopran Hydrochloride
(C18H23N9O4S3: 525.63).
セフォゾプラン塩酸塩
Method of Preparation Prepare as directed under Injec-
tion, with Cefotiam Hydrochloride.
Description Cefotiam Hydrochloride for Injection occurs
as a white to light yellow powder.
solution of Cefotiam Hydrochloride for Injection (1 in
tometry <2.24>: it exhibits a maximum between 257 nm and C19H17N9O5S2.HCl: 551.99
261 nm. (6R,7R)-7-[(Z )-2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-
(2) Determine the 1H spectrum of a solution of Cefotiam (methoxyimino)acetylamino]-3-(1H-
Hydrochloride for Injection in heavy water for nuclear mag- imidazo[1,2-b]pyridazin-4-ium-1-ylmethyl)-8-oxo-
netic resonance spectroscopy (1 in 10) as directed under 5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate
Nuclear Magnetic Resonance Spectroscopy <2.21>, using so- monohydrochloride
dium 3-trimethylsilylpropanesulfonate for nuclear magnetic [113359-04-9, Cefozopran]
resonance spectroscopy as an internal reference compound:
it exhibits a single signal A between d 2.7 ppm and d 3.0 Cefozopran Hydrochloride contains not less than
ppm, and a single signal B at around d 6.5 ppm. The ratio of 860 mg (potency) and not more than 960 mg (potency)
the integrated intensity of each signal, A:B, is about 6:1. per mg, calculated on the anhydrous basis. The po-
tency of Cefozopran Hydrochloride is expressed as
mass (potency) of cefozopran (C19H17N9O5S2: 515.53).
amount of Cefotiam Hydrochloride for Injection, equivalent
to 0.5 g (potency), in 5 mL of water is between 5.7 and 7.2. Description Cefozopran Hydrochloride occurs as white to
pale yellow, crystals or crystalline powder.
It is freely soluble in dimethylsulfoxide and in formamide,
of Cefotiam Hydrochloride for Injection, equivalent to 1.0 g
slightly soluble in water, in methanol and in ethanol (95),
(potency) of Cefotiam Hydrochloride, in 10 mL of water:
and practically insoluble in acetonitrile and diethyleter.
the solution is clear, and the absorbance of this solution,
determined at 450 nm 10 minutes after dissolving as directed Identification (1) Dissolve 0.02 g of Cefozopran Hydro-
under Ultraviolet-visible Spectrophotometry <2.24>, is not chloride in 10 mL of water, add 1 mL of a solution of hy-
more than 0.20. droxylammonium chloride (1 in 10) and 2 mL of sodium hy-
droxide TS, allow to stand for 5 minutes, then add 3 mL of 1
mol/L hydrochloric acid TS and 3 drops of iron (III) chlo-
um, 609C, 3 hours).
ride TS, and mix: a red-purple color develops.
Bacterial endotoxins <4.01> Less than 0.125 EU/mg (po- (2) Determine the absorption spectra of solutions of
tency). Cefozopran Hydrochloride and Cefozopran Hydrochloride
RS in a mixture of sodium chloride TS and methanol (3:2)
photometry <2.24>, and compare these spectra: both spectra
Foreign insoluble matter <6.06> Perform the test according exhibit similar intensities of absorption at the same wave-
to Method 2: it meets the requirement. lengths.
Cefozopran Hydrochloride in deuterated dimethylsulfoxide
ment.
for nuclear magnetic resonance spectroscopy (1 in 20) as
Sterility <4.06> Perform the test according to the Mem- directed under Nuclear Magnetic Resonance Spectroscopy
brane filtration method: it meets the requirement. <2.21>, using tetramethylsilane for nuclear magnetic reso-
JP XVII Official Monographs / Cefozopran Hydrochloride for Injection 641
nance spectroscopy as an internal reference compound: it Flow rate: Adjust so that the retention time of cefozopran
exhibits a single signal A at around d 3.9 ppm, a double is about 9 minutes.
signal B at around d 5.2 ppm, and a quartet signal C at System suitability—
around d 8.0 ppm, and the ratio of integrated intensity of System performance: When the procedure is run with 10
each signal, A:B:C, is about 3:1:1. mL of the standard solution under the above operating con-
(4) Dissolve 0.01 g of Cefozopran Hydrochloride in 1 ditions, cefozopran and the internal standard are eluted in
mL of water and 2 mL of acetic acid (100), add 2 drops of this order with the resolution between these peaks being not
silver nitrate TS, and mix: a white turbidity is formed. less than 10.
lated on the anhydrous basis, a mixture of sodium chloride
TS and methanol (3:2), 5000 mL).
of the peak area of cefozopran to that of the internal stand-
D : －73 – －789(0.1 g calculated ard is not more than 1.0z.
on the anhydrous basis, a mixture of sodium chloride TS and
methanol (3:2), 10 mL, 100 mm).
Purity (1) Clarity and color of solution—Being specified
separately when the drug is granted approval based on the
Law. Cefozopran Hydrochloride for
Cefozopran Hydrochloride according to Method 2, and Injection
perform the test. Prepare the control solution with 2.0 mL
注射用セフォゾプラン塩酸塩
of Standard Lead Solution (not more than 10 ppm).
(3) Arsenic—Being specified separately when the drug is
granted approval based on the Law. Cefozopran Hydrochloride for Injection is a prepa-
(4) Related substances—Being specified separately when ration for injection which is dissolved before use.
the drug is granted approval based on the Law. It contains not less than 90.0z and not more
than 115.0z of the labeled potency of cefozopran
(C19H17N9O5S2: 515.53).
determination and methanol for water determination (2:1) Method of Preparation Prepare as directed under the In-
instead of methanol for water determination). jections, with Cefozopran Hydrochloride.
Residue on ignition Being specified separately when the Description Cefozopran Hydrochloride for Injection oc-
drug is granted approval based on the Law. curs as a white to light yellow, powder or masses.
Bacterial endotoxins <4.01> Less than 0.05 EU/mg (po- Identification (1) Determine the absorption spectrum of a
tency). solution of Cefozopran Hydrochloride for Injection (1 in
Assay Weigh accurately an amount of Cefozopran Hydro-
tometry <2.24>: it exhibits a maximum between 236 nm and
chloride and Cefozopran Hydrochloride RS, equivalent to
241 nm.
about 50 mg (potency), and dissolve each in the mobile phase
(2) To 50 mg of Cefozopran Hydrochloride for Injection
to make exactly 50 mL. Pipet 10 mL each of these solutions,
add 0.8 mL of deuterated dimethylsulfoxide for nuclear
add exactly 10 mL of the internal standard solution and the
magnetic resonance spectroscopy, and filter after shaking,
mobile phase to make 25 mL, and use these solutions as the
and determine the 1H spectrum of the filtrate as directed
sample solution and the standard solution, respectively. Per-
under Nuclear Magnetic Resonance Spectroscopy <2.21>,
using tetramethylsilane for nuclear magnetic resonance
spectroscopy as an internal reference compound: it exhibits a
single signal A at around d 3.9 ppm, a double signal B at
the ratios, QT and QS, of the peak area of cefozopran to that
around d 5.0 ppm, and a quartet signal C at around d 8.0
of the internal standard.
ppm. The ratio of the integrated intensity of each signal,
Amount [ mg (potency)] of cefozopran (C19H17N9O5S2) A:B:C, is about 3:1:1.
＝ MS × QT/QS × 1000
pH <2.54> Dissolve an amount of Cefozopran Hydrochlo-
MS: Amount [mg (potency)] of Cefozopran Hydrochlo- ride for Injection, equivalent to 0.5 g (potency) of
ride RS taken Cefozopran Hydrochloride, in 5 mL of water: the pH of this
solution is between 7.5 and 9.0.
Internal standard solution—A solution of 2,4-dihydroxyben-
zoic acid in the mobile phase (1 in 1250). Purity (1) Clarity and color of solution—Dissolve an
Operating conditions— amount of Cefozopran Hydrochloride for Injection, equiva-
Detector: An ultraviolet absorption photometer (wave- lent to 1 g (potency) of Cefozopran Hydrochloride, in 10 mL
length: 254 nm). of water: the solution is clear and has no more color than
Column: A stainless steel column 4.6 mm in inside diame- Matching Fluid N.
ter and 15 cm in length, packed with octadecylsilanized silica (2) Related substances—Being specified separately when
gel for liquid chromatography (5 mm in particle diameter). the drug is granted approval based on the Law.
259 C.
Mobile phase: Mix 0.366 g of diethylamine with water to
make 1000 mL, and add 60 mL of acetonitrile and 5 mL of
instead of methanol for water determination).
acetic acid (100).
642 Cefpiramide Sodium / Official Monographs JP XVII
Bacterial endotoxins <4.01> Less than 0.05 EU/mg (po- water, sparingly soluble in methanol, and slightly soluble in
tency). ethanol (95).
Uniformity of dosage units <6.02> It meets the requirement Identification (1) Determine the absorption spectrum of a
of the Mass variation test. solution of Cefpiramide Sodium in 0.05 mol/L phosphate
buffer solution (pH 7.0) (1 in 50,000) as directed under Ul-
Foreign insoluble matter <6.06> It meets the requirement.
Insoluble particulate matter <6.07> It meets the require- spectrum with the Reference Spectrum: both spectra exhibit
ment. similar intensities of absorption at the same wavelengths.
(2) Determine the 1H spectrum of a solution of Cef-
piramide Sodium in deuterated dimethylsulfoxide for
nuclear magnetic resonance spectroscopy (1 in 10) as directed
Assay Weigh accurately the mass of the contents of not less under Nuclear Magnetic Resonance Spectroscopy <2.21>,
than 10 Cefozopran Hydrochloride for Injection. Weigh ac- using tetramethylsilane for nuclear magnetic resonance
curately an amount of the contents, equivalent to about 0.5 g spectroscopy as an internal reference compound: it exhibits
(potency) of Cefozopran Hydrochloride, and add water to single signals, A, B and C, at around d 2.3 ppm, at around
make exactly 100 mL. Pipet 2 mL of this solution, add ex- d 3.9 ppm and at around d 8.2 ppm, respectively. The ratio
actly 10 mL of the internal standard solution, add the mobile of the integrated intensity of each signal, A:B:C, is about
phase to make 25 mL, and use this solution as the sample so- 3:3:1.
lution. Separately, weigh accurately an amount of (3) Cefpiramide Sodium responds to the Qualitative
Cefozopran Hydrochloride RS, equivalent to about 50 mg Tests <1.09> (1) for sodium salt.
(potency), and dissolve in the mobile phase to make exactly
D : －33 – －409(0.2 g calculated
50 mL. Pipet 10 mL of this solution, add exactly 10 mL of
on the anhydrous basis, 0.05 mol/L phosphate buffer solu-
the internal standard solution, add the mobile phase to make
tion (pH 7.0), 10 mL, 100 mm).
25 mL, and use this solution as the standard solution. Pro-
ceed as directed in the Assay under Cefozopran Hydrochlo- pH <2.54> The pH of a solution obtained by dissolving
ride. 2.0 g of Cefpiramide Sodium in 20 mL of water is between
5.5 and 8.0.
Amount [mg (potency)] of cefozopran (C19H17N9O5S2)
＝ MS × QT/QS × 10 Purity (1) Clarity and color of solution—Dissolve 1.0 g
of Cefpiramide Sodium in 10 mL of 0.05 mol/L phosphate
MS: Amount [mg (potency)] of Cefozopran Hydrochlo-
buffer solution (pH 7.0): the solution is clear, and colorless
ride RS taken
or light yellow.
Internal standard solution—A solution 2,4-dihydroxyben- (2) Heavy metals <1.07>—Proceed with 1.0 g of Cef-
zoic acid in the mobile phase (1 in 1250). piramide Sodium according to Method 2, and perform the
(3) Related substances—Weigh accurately about 25 mg
of Cefpiramide Sodium, dissolve in 0.03 mol/L phosphate
buffer solution (pH 7.5) to make exactly 50 mL, and use this
Cefpiramide Sodium about 25 mg of 1-methyl-1H-tetrazole-5-thiol for liquid
chromatography, previously dried in a desiccator (in vacu-
セフピラミドナトリウム
um, silica gel) for 2 hours, and an amount of Cefpiramide
RS, equivalent to about 75 mg (potency), dissolve them in
0.03 mol/L phosphate buffer solution (pH 7.5) to make ex-
actly 100 mL. Pipet 2 mL of this solution, add 0.03 mol/L
test with exactly 5 mL of the sample solution and standard
C25H23N8NaO7S2: 634.62
Monosodium (6R,7R)-7-{(2R)-2-[(4-hydroxy-6-
the amount of 1-methyl-1H-tetrazole-5-thiol, each of the
methylpyridine-3-carbonyl)amino]-2-(4-
other related substances and the total of the other related
hydroxyphenyl)acetylamino}-3-(1-methyl-1H-tetrazol-
substances by the following equations: the amount of 1-
5-ylsulfanylmethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-
methyl-1H-tetrazole-5-thiol, each of the other related sub-
2-ene-2-carboxylate
stances and the total of the other related substances are not
[74849-93-7]
more than 1.0z, not more than 1.5z and not more than
4.0z, respectively.
Cefpiramide Sodium contains not less than 900 mg
(potency) and not more than 990 mg (potency) per mg, Amount (z) of 1-methyl-1H-tetrazole-5-thiol (C2H4N4S)
calculated on the anhydrous basis. The potency of ＝ MSa/MT × ATa/ASa
Cefpiramide Sodium is expressed as mass (potency) of
Amount (z) of each of other related substances
cefpiramide (C25H24N8O7S2: 612.64). ＝ MSb/MT × ATc/ASb
Description Cefpiramide Sodium occurs as white to yellow-
MSa: Amount (mg) of 1-methyl-1H-tetrazole-5-thiol taken
ish white powder.
MSb: Amount [mg (potency)] of Cefpiramide RS taken
It is very soluble in dimethylsulfoxide, freely soluble in
JP XVII Official Monographs / Cefpirome Sulfate 643
MT: Amount (mg) of Cefpiramide Sodium taken Mobile phase: A mixture of 0.01 mol/L phosphate buffer
ASa: Peak area of 1-methyl-1H-tetrazole-5-thiol from the solution (pH 6.8), acetonitrile, methanol and tetrahydrofu-
standard solution ran (22:1:1:1).
ASb: Peak area of cefpiramide from the standard solution Flow rate: Adjust so that the retention time of cefpiramide
ATa: Peak area of 1-methyl-1H-tetrazole-5-thiol from the is about 7 minutes.
sample solution System suitability—
ATc: Area of each peak other than 1-methyl-1H-tetrazole- System performance: When the procedure is run with 5 mL
5-thiol and cefpiramide from the sample solution of the standard solution under the above operating condi-
tions, cefpiramide and the internal standard are eluted in this
than 7.
length: 254 nm).
and 30 cm in length, packed with octylsilanized silica gel for
liquid chromatography (10 mm in particle diameter).
peak area of cefpiramide to that of the internal standard is
not more than 2.0z.
259 C.
Mobile phase: A mixture of 0.03 mol/L phosphate buffer Containers and storage Containers—Tight containers.
solution (pH 7.5) and methanol (3:1). Storage—Light-resistant, and at a temperature not
Flow rate: Adjust so that the retention time of cefpiramide exceeding 59C.
retention time of cefpiramide. Cefpirome Sulfate
Test for required detectability: Measure exactly 5 mL of セフピロム硫酸塩
the standard solution, and add 0.03 mol/L phosphate buffer
solution (pH 7.5) to make exactly 50 mL. Confirm that the
peak area of 1-methyl-1H-tetrazole-5-thiol obtained from
tained from 5 mL of the standard solution.
System performance: Dissolve 25 mg of Cefpiramide RS
and 7 mg of cinnamic acid in the mobile phase to make 50
C22H22N6O5S2.H2SO4: 612.66
mL. When the procedure is run with 5 mL of this solution
under the above operating conditions, cinnamic acid and cef-
(methoxyimino)acetylamino]-3-(6,7-dihydro-5H-
piramide are eluted in this order with the resolution between
cyclopenta[b]pyridinium-1-ylmethyl)-8-oxo-5-thia-1-
azabicyclo[4.2.0]oct-2-ene-2-carboxylate monosulfate
[98753-19-6]
Cefpirome Sulfate contains not less than 760 mg
of 1-methyl-1H-tetrazole-5-thiol is not more than 2.0z.
(potency) per mg, calculated on the anhydrous basis.
Water <2.48> Not more than 7.0z (0.35 g, volumetric The potency of Cefpirome Sulfate is expressed as mass
titration, direct titration). (potency) of cefpirome (C22H22N6O5S2: 514.58).
Assay Weigh accurately an amount of Cefpiramide So- Description Cefpirome Sulfate occurs as a white to pale
dium and Cefpiramide RS, equivalent to about 50 mg (po- yellowish white crystalline powder, and has a slight, charac-
tency), add exactly 5 mL of the internal standard solution to teristic odor.
dissolve, then add the mobile phase to make 100 mL, and It is soluble in water, and practically insoluble in ethanol
use these solutions as the sample solution and standard solu- (95) and in diethyl ether.
tion. Perform the test with 5 mL each of the sample solution It is hygroscopic.
Identification (1) Dissolve 10 mg of Cefpirome Sulfate in
raphy <2.01> according to the following conditions, and cal-
2 mL of water, add 3 mL of hydroxylammonium hydrochlo-
culate the ratios, QT and QS, of the peak area of cefpiramide
ride-ethanol TS, allow to stand for 5 minutes, add 1 mL of
to that of the internal standard.
acidic ammonium iron (III) sulfate TS, and shake: a red-
Amount [ mg (potency)] of cefpiramide (C25H24N8O7S2) brown color develops.
＝ MS × QT/QS × 1000 (2) Dissolve 1 mg of Cefpirome Sulfate in 4 mL of water,
add 1 mL of dilute hydrochloric acid while cooling in ice,
MS: Amount [mg (potency)] of Cefpiramide RS taken
add 1 mL of a freshly prepared solution of sodium nitrite (1
Internal standard solution—A solution of 4-dimethylamino- in 100), and allow to stand for 2 minutes. Add 1 mL of am-
antipyrine (1 in 100). monium amidosulfuric acid TS while cooling in ice bath,
Operating conditions— allow to stand for 1 minute, and add 1 mL of a solution of
Detector: An ultraviolet absorption photometer (wave- N-1-naphthylethylene dihydrochloride (1 in 1000): a purple
length: 254 nm). color develops.
Column: A stainless steel column 4 mm in inside diameter (3) Take 5 mg of Cefpirome Sulfate, dissolve in 1 mL of
and 30 cm in length, packed with octylsilanized silica gel for ethanol (95) and 1 mL of water, add 100 mg of 1-chloro-2,4-
liquid chromatography (10 mm in particle diameter). dinitrobenzene, and heat on a water bath for 5 minutes.
Column temperature: A constant temperature of about After cooling, add 2 or 3 drops of a solution of sodium hy-
259 C. droxide (1 in 10) and 3 mL of ethanol (95): a red-brown
644 Cefpodoxime Proxetil / Official Monographs JP XVII
color develops. Operating conditions—
(4) Determine the absorption spectra of solutions of Detector: An ultraviolet absorption photometer (wave-
Cefpirome Sulfate and Cefpirome Sulfate RS in 0.01 mol/L length: 270 nm).
hydrochloric acid TS (1 in 50,000) as directed under Ultra- Column: A stainless steel column 4.6 mm in inside diame-
violet-visible Spectrophotometry <2.24>, and compare the ter and 25 cm in length, packed with octadecylsilanized silica
spectra: both spectra exhibit similar intensities of absorption gel for liquid chromatography (5 mm in particle diameter).
at the same wavelengths. Column temperature: A constant temperature of about
(5) Determine the 1H spectrum of a solution of Cefpi- 259C.
rome Sulfate in heavy water for nuclear magnetic resonance Mobile phase: Dissolve 3.45 g of ammonium dihydrogen-
spectroscopy (1 in 25) as directed under Nuclear Magnetic phosphate in 1000 mL of water, and adjust the pH to 3.3
Resonance Spectroscopy <2.21>, using sodium 3-trimethyl- with phosphoric acid. To 800 mL of this solution add 100
silylpropanesulfonate for nuclear magnetic resonance spec- mL of acetonitrile.
troscopy as an internal reference compound: it exhibits a Flow rate: Adjust so that the retention time of cefpirome
single signal A at around d 4.1 ppm, a double signal B at is about 7.5 minutes.
around d 5.9 ppm, a single signal C at around d 7.1 ppm, System suitability—
and a multiple signal D at around d 7.8 ppm. The ratio of System performance: When the procedure is run with 20
integrated intensity of each signal, A:B:C:D, is about mL of the standard solution under the above operating con-
3:1:1:1. ditions, the number of theoretical plates of the peak of cefpi-
(6) A solution of Cefpirome Sulfate (1 in 250) responds rome is not less than 3600.
to the Qualitative Tests <1.09> (1) for sulfate salt. System repeatability: When the test is repeated 5 times
lated on the anhydrous basis, 0.01 mol/L hydrochloric acid
areas of cefpirome is not more than 1.0z.
TS, 2500 mL).
D : －27 – －339 (50 mg calcu-
Storage—At a temperature between 29C and 89C.
lated on the anhydrous basis, a solution prepared by addi-
tion of water to 25 mL of acetonitrile to make 50 mL, 20
mL, 100 mm).
Cefpodoxime Proxetil
pH <2.54> Dissolve 0.1 g of Cefpirome Sulfate in 10 mL of
water: the pH of the solution is between 1.6 and 2.6. セフポドキシムプロキセチル
Purity (1) Clarity and color of solution—Being specified
separately when the drug is granted approval based on the
Law.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Cefpi-
rome Sulfate according to Method 2, and perform the test.
(3) Arsenic—Being specified separately when the drug is
C21H27N5O9S2: 557.60
(1RS )-1-[(1-Methylethoxy)carbonyloxy]ethyl
(6R,7R)-7-[(Z )-2-(2-aminothiazol-4-yl)-2-
Water <2.48> Not more than 2.5z (0.5 g, volumetric titra- (methoxyimino)acetylamino]-3-methoxymethyl-8-oxo-5-
tion, direct titration). thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate
[87239-81-4]
Residue on ignition Being specified separately when the
Cefpodoxime Proxetil contains not less than 706 mg
Bacterial endotoxins <4.01> Less than 0.10 EU/mg (po- (potency) and not more than 774 mg (potency) per mg,
tency). calculated on the anhydrous basis. The potency of
Cefpodoxime Proxetil is expressed as mass (potency)
Assay Weigh accurately an amount of Cefpirome Sulfate
of cefpodoxime (C15H17N5O6S2: 427.46).
and Cefpirome Sulfate RS, equivalent to about 50 mg (po-
tency), dissolve each in water to make exactly 100 mL. Pipet Description Cefpodoxime Proxetil occurs as a white to
5 mL of these solutions, add each in water to make exactly light brownish white powder.
20 mL, and use these solutions as the sample solution and It is very soluble in acetonitrile, in methanol and in chlo-
standard solution. Perform the test with exactly 20 mL of the roform, freely soluble in ethanol (99.5), and very slightly
sample solution and standard solution as directed under soluble in water.
solution of Cefpodoxime Proxetil in acetonitrile (3 in
cefpirome in each solution.
Amount [ mg (potency)] of cefpirome (C22H22N6O5S2) tometry <2.24>, and compare the spectrum with the Refer-
＝ MS × AT/AS × 1000 ence Spectrum or the spectrum of a solution of Cefpodoxime
Proxetil RS prepared in the same manner as the sample solu-
MS: Amount [mg (potency)] of Cefpirome Sulfate RS
taken
JP XVII Official Monographs / Cefpodoxime Proxetil 645
podoxime Proxetil as directed in the potassium bromide disk solution for system suitability test. Pipet 2 mL of the solu-
method under Infrared Spectrophotometry <2.25>, and com- tion for system suitability test, and add the mixture of water,
pare the spectrum with the Reference Spectrum or the spec- acetonitrile and acetic acid (100) (99:99:2) to make exactly
trum of Cefpodoxime Proxetil RS: both spectra exhibit simi- 100 mL. Confirm that the peak areas of the isomer A and
lar intensities of absorption at the same wave numbers. the isomer B of cefpodoxime proxetil obtained from 20 mL
(3) Determine the 1H spectrum of a solution of Cef- of this solution are equivalent to 1.4 to 2.6z of them ob-
podoxime Proxetil in deuterochloroform for nuclear mag- tained from 20 mL of the solution for system suitability test,
netic resonance spectroscopy (1 in 10) as directed under respectively.
Nuclear Magnetic Resonance Spectroscopy <2.21>, using System performance: When the procedure is run with 20
tetramethylsilane for nuclear magnetic resonance spectros- mL of the solution for system suitability test under the above
copy as an internal reference compound: it exhibits double operating conditions, the isomer A and the isomer B of cef-
signals, A and B, at around d 1.3 ppm and at around d 1.6 podoxime proxetil are eluted in this order with the resolution
ppm, and single signals, C and D, at around d 3.3 ppm and between these peaks being not less than 6.
at around d 4.0 ppm. The ratio of the integrated intensity of System repeatability: When the test is repeated 5 times
these signals, A:B:C:D, is about 2:1:1:1. with 20 mL of the solution for system suitability test under
D : ＋24.0 – ＋31.49(0.1 g calcu-
tion of the peak area of the isomer A and the isomer B of
lated on the anhydrous basis, acetonitrile, 20 mL, 100 mm).
cefpodoxime proxetil is not more than 2.0z.
Cefpodoxime Proxetil according to Method 2, and perform
ard Lead Solution (not more than 20 ppm). Residue on ignition <2.44> Not more than 0.2z (1 g).
(2) Related substances—Dissolve 50 mg of Cefpodoxime
Isomer ratio Perform the test with 5 mL of the sample
Proxetil in 50 mL of a mixture of water, acetonitrile and
solution obtained in the Assay as directed under Liquid
acetic acid (100) (99:99:2), and use this solution as the sam-
ple solution. Perform the test with 20 mL of the sample solu-
tions, and determine the peak areas of the two isomers of
cefpodoxime proxetil, Aa, for the isomer having the smaller
retention time, and Ab, for the isomer having the larger
area by the automatic integration method, and calculate the
retention time, by the automatic integration method: Ab/
amounts of them by the area percentage method: the amount
(Aa ＋ Ab) is between 0.50 and 0.60.
of the peak, having the relative retention time of about 0.8
to the isomer B of cefpodoxime proxetil, is not more than
2.0z, the amount of the peak other than cefpodoxime
Assay.
proxetil is not more than 1.0z, and the total amount of the
peaks other than cefpodoxime proxetil is not more than
6.0z.
of the standard solution obtained in the Assay under the
above operating conditions, the internal standard, the
isomer A and the isomer B of cefpodoxime proxetil are
length: 254 nm).
eluted in this order with the resolution between the peaks of
the isomers being not less than 4.
with 5 mL of the standard solution obtained in the Assay
under the above operating conditions, the relative standard
229 C.
deviation of the ratio of the peak area of the isomer B of cef-
Mobile phase A: A mixture of water, methanol and a
podoxime proxetil to that of the internal standard is not
solution of formic acid (1 in 50) (11:8:1).
more than 1.0z.
Mobile phase B: A mixture of methanol and a solution of
formic acid (1 in 50) (19:1). Assay Weigh accurately an amount of Cefpodoxime Prox-
Flowing of mobile phase: Control the gradient by mixing etil and Cefpodoxime Proxetil RS, equivalent to about 60
the mobile phases A and B as directed in the following table. mg (potency), dissolve in 80 mL of acetonitrile, add exactly 4
mL of the internal standard solution, add acetonitrile to
Time after injection Mobile phase A Mobile phase B make 100 mL, and use these solutions as the sample solution
of sample (min) (volz) (volz) and standard solution. Perform the test with 5 mL each of
0 – 65 95 5 Liquid Chromatography <2.01> according to the following
65 – 145 95 → 15 5 → 85 conditions, and calculate the ratios, QT1, QS1, QT2 and QS2,
145 – 155 15 85 of the areas of the two peaks of the isomers of cefpodoxime
proxetil to the peak area of the internal standard.
Flow rate: 0.7 mL per minute (the retention time of the Amount [ mg (potency)] of cefpodoxime (C15H17N5O6S2)
isomer B of cefpodoxime proxetil is about 60 minutes). ＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 1000
Time span of measurement: For 155 minutes after injec-
tion, beginning after the solvent peak. MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS
System suitability— taken
Test for required detectability: To 5 mL of the sample so- Internal standard solution—Dissolve 0.3 g of ethyl parahy-
lution add a mixture of water, acetonitrile and acetic acid droxybenzoate in a solution of citric acid in acetonitrile (1 in
(100) (99:99:2) to make 200 mL, and use this solution as the 2000) to make 100 mL.
646 Cefpodoxime Proxetil for Syrup / Official Monographs JP XVII
Operating conditions— Then, proceed as directed in the Assay under Cefpodoxime
Detector: An ultraviolet absorption photometer (wave- Proxetil.
length: 240 nm).
Amount [mg (potency)] of cefpodoxime (C15H17N5O6S2)
＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 2
gel for liquid chromatography (5 mm in particle diameter). MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS
Column temperature: A constant temperature of about taken
409 C.
Internal standard solution—Dissolve 0.2 g of ethyl parahy-
Mobile phase: A mixture of water and methanol (11:9).
droxybenzoate in a mixture of water, acetonitrile and acetic
acid (100) (99:99:2) to make 300 mL.
standard is about 11 minutes.
System suitability— Dissolution <6.10> When the test is performed at 50 revolu-
System performance: When the procedure is run with 5 mL tions per minute according to the Paddle method, using 900
of the standard solution under the above operating condi- mL of water as the dissolution medium, the dissolution rates
tions, the internal standard, the isomer A and the isomer B in 15 minutes of Cefpodoxime Proxetil for Syrup is not less
are eluted in this order with the resolution between these than 85z.
peaks being not less than 4. Start the test with an accurately weighed amount of Cef-
System repeatability: When the test is repeated 5 times podoxime Proxetil for Syrup, equivalent to about 50 mg (po-
with 5 mL of the standard solution under the above operating tency) of Cefpodoxime Proxetil, withdraw not less than 20
conditions, the relative standard deviation of the ratios of mL of the medium at the specified minute after starting the
the peak area of the isomer B of cefpodoxime proxetil to test, and filter through a membrane filter with a pore size
that of the internal standard is not more than 1.0z. not exceeding 0.5 mm. Discard the first 10 mL of the filtrate,
pipet 5 mL of the subsequent filtrate, add a solution of citric
acid monohydrate in the mobile phase (1 in 2000) to make
Separately, weigh accurately an amount of Cefpodoxime
Cefpodoxime Proxetil for Syrup Proxetil RS, equivalent to about 22 mg (potency), dissolve in
a solution of citric acid monohydrate in the mobile phase (1
シロップ用セフポドキシムプロキセチル
in 2000) to make exactly 100 mL. Pipet 5 mL of this solu-
tion, add a solution of citric acid monohydrate in the mobile
Cefpodoxime Proxetil for Syrup is a preparation for phase (1 in 2000) to make exactly 100 mL, and use this solu-
syrups which is suspended before use. tion as the standard solution. Perform the test with exactly
It contains not less than 93.0z and not more 10 mL each of the sample solution and standard solution as
than 107.0z of the labeled potency of cefpodoxime directed under Liquid Chromatography <2.01> according to
(C15H17N5O6S2: 427.46). the following conditions, and determine the areas, ATa and
ASa, of the one peak which appears at the retention time of
Method of preparation Prepare as directed under Syrups,
about 24 minutes among the two peaks obtainable from cef-
with Cefpodoxime Proxetil.
podoxime proxetil, and the areas, ATb and ASb, of the peak
Identification To an amount of Cefpodoxime Proxetil for which appears at the retention time of about 30 minutes, in
Syrup, equivalent to 15 mg (potency) of Cefpodoxime Prox- each solution.
etil, add 10 mL of 0.1 mol/L phosphate buffer solution (pH
8.0), treat with ultrasonic waves for 5 minutes while occa-
of cefpodoxime proxetil (C21H27N5O9S2)
sional shaking. Then, add 20 mL of ethyl acetate, shake for
＝ MS/MT × (ATa ＋ ATb)/(ASa ＋ ASb) × 1/C × 225
5 minutes, and centrifuge. Take 3 mL of the supernatant liq-
uid, evaporate the ethyl acetate by warming at 409C under MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS
reduced pressure. Dissolve the residue in acetonitrile to make taken
200 mL, and determine the absorption spectrum of this solu- MT: Amount (g) of Cefpodoxime Proxetil for Syrup taken
tion as directed under Ultraviolet-visible Spectrophotometry C: Labeled amount [mg (potency)] of cefpodoxime prox-
<2.24>: it exhibits a maximum between 232 nm and 236 nm. etil (C21H27N5O9S2) in 1 g
Uniformity of dosage units <6.02> Perform the test accord- Operating conditions—
ing to the following method: Cefpodoxime Proxetil for Detector, column, column temperature, and mobile phase:
Syrup in single-dose packages meet the requirement of the Proceed as directed in the operating conditions in the Assay
Content uniformity test. under Cefpodoxime Proxetil.
To the total content of 1 package of Cefpodoxime Proxetil Flow rate: Adjust so that the retention time of the peak,
for Syrup add exactly 30 mL of the internal standard solu- which elutes faster among the two peaks obtained from cef-
tion, treat with ultrasonic waves for 10 minutes while occa- podoxime proxetil, is about 24 minutes.
sional shaking, and centrifuge. Take 3 mL of the superna- System suitability—
tant liquid, add a mixture of water, acetonitrile and acetic System performance: When the procedure is run with 10
acid (100) (99:99:2) to make 20 mL, and use this solution as mL of the standard solution under the above operating con-
the sample solution. Separately, weigh accurately an amount ditions, the resolution between the two peaks obtained from
of Cefpodoxime Proxetil RS, equivalent to about 50 mg (po- cefpodoxime proxetil is not less than 4.
tency), dissolve in a suitable amount of a mixture of water, System repeatability: When the test is repeated 6 times
acetonitrile and acetic acid (100) (99:99:2), add exactly 15 with 10 mL of the standard solution under the above operat-
mL of the internal standard solution, then add a mixture of ing conditions, the relative standard deviation of the sum of
water, acetonitrile and acetic acid (100) (99:99:2) to make the areas of the two peaks obtained from cefpodoxime prox-
100 mL, and use this solution as the standard solution. etil is not more than 2.0z.
JP XVII Official Monographs / Cefpodoxime Proxetil Tablets 647
Assay Weigh accurately an amount of powdered Cef- ard solution. Then, proceed as directed in the Assay under
podoxime Proxetil for Syrup, equivalent to about 0.1 g Cefpodoxime Proxetil.
(potency) of Cefpodoxime Proxetil, add exactly 30 mL of
the internal standard solution, treat with ultrasonic waves
＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 10/V
for 10 minutes while occasional shaking, and centrifuge.
Take 3 mL of the supernatant liquid, add a mixture of MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS
water, acetonitrile and acetic acid (100) (99:99:2) to make 20 taken
weigh accurately an amount of Cefpodoxime Proxetil RS,
equivalent to about 50 mg (potency), dissolve in a suitable
acid (100) (99:99:2) to make 100 mL.
amount of a mixture of water, acetonitrile and acetic acid
(100) (99:99:2), add exactly 15 mL of the internal standard Dissolution <6.10> When the test is performed at 50 revolu-
solution, then add a mixture of water, acetonitrile and acetic tions per minute according to the Paddle method, using 900
acid (100) (99:99:2) to make 100 mL, and use this solution as mL of water as the dissolution medium, the dissolution rate
the standard solution. Then, proceed as directed in the Assay in 45 minutes of Cefpodoxime Proxetil Tablets is not less
under Cefpodoxime Proxetil. than 70z.
Start the test with 1 tablet of Cefpodoxime Proxetil
＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 2
MS: Amount [mg (potency)] of Cefpodoxime Proxetil membrane filter with a pore size not exceeding 0.5 mm. Dis-
RS taken card the first 10 mL of the filtrate, pipet V mL of the subse-
quent filtrate, add a solution of citric acid monohydrate in
the mobile phase (1 in 2000) to make exactly V? mL so that
each mL contains about 11 mg (potency) of Cefpodoxime
acid (100) (99:99:2) to make 300 mL.
Proxetil, and use this solution as the sample solution. Sepa-
Containers and storage Containers—Tight containers. rately, weigh accurately an amount of Cefpodoxime Proxetil
RS, equivalent to about 22 mg (potency), and dissolve in a
solution of citric acid monohydrate in the mobile phase (1 in
Cefpodoxime Proxetil Tablets 2000) to make exactly 100 mL. Pipet 5 mL of this solution,
add a solution of citric acid monohydrate in the mobile
セフポドキシムプロキセチル錠 phase (1 in 2000) to make exactly 100 mL, and use this solu-
Cefpodoxime Proxetil Tablets contain not less than
93.0z and not more than 107.0z of the labeled po-
the following conditions, and determine the areas of sepa-
tency of cefpodoxime (C15H17N5O6S2: 427.46).
rated two peaks, one has the retention time of about 24
Method of preparation Prepare as directed under Tablets, minutes, ATa and ASa, and another one has the retention time
with Cefpodoxime Proxetil. of about 30 minutes, ATb and ASb, in each solution.
Identification Powder Cefpodoxime Proxetil Tablets. To a Dissolution rate (z) with respect to the labeled amount of
portion of the powder, equivalent to 65 mg (potency) of Cef- cefpodoxime proxetil (C21H27N5O9S2)
podoxime Proxetil, add 25 mL of acetonitrile, shake thor- ＝ MS × (ATa ＋ ATb)/(ASa ＋ ASb) × V?/V × 1/C × 45
oughly, and centrifuge. To 2 mL of the supernatant liquid
MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS
add acetonitrile to make 50 mL. To 5 mL of this solution
taken
add acetonitrile to make 50 mL. Determine the absorption
C: Labeled amount [mg (potency)] of cefpodoxime prox-
spectrum of this solution as directed under Ultraviolet-
etil (C21H27N5O9S2) in 1 tablet
tween 232 nm and 236 nm. Operating conditions—
length: 240 nm).
To 1 tablet of Cefpodoxime Proxetil Tablets, add exactly
20 mL of a mixture of water, acetonitrile and acetic acid
(100) (99:99:2), agitate with the aid of ultrasonic waves for
409C.
10 minutes, and filter through a membrane filter with a pore
Mobile phase: A mixture of water and methanol (11:9).
Flow rate: Adjust so that the retention time of one of the
trate, pipet V mL of the subsequent filtrate, equivalent to 30
two peaks that elutes firster is about 24 minutes.
mg (potency) of Cefpodoxime Proxetil, add exactly 6 mL of
the internal standard solution, then add a mixture of water,
acetonitrile and acetic acid (100) (99:99:2) to make 50 mL,
mL of the standard solution under the above operating
conditions, the resolution between the two peaks of cef-
weigh accurately an amount of Cefpodoxime Proxetil RS,
podoxime proxetil is not less than 4.
equivalent to about 60 mg (potency), dissolve in 60 mL of a
mixture of water, acetonitrile and acetic acid (100) (99:99:2),
add exactly 12 mL of the internal standard solution, then
ing conditions, the relative standard deviation of the total
add a mixture of water, acetonitrile and acetic acid (100)
area of the two peaks of cefpodoxime proxetil is not more
(99:99:2) to make 100 mL, and use this solution as the stand-
than 2.0z.
648 Cefroxadine Hydrate / Official Monographs JP XVII
Assay Weigh accurately the mass of not less than 20 Cef- Cefroxadine RS prepared in the same manner as the sample
podoxime Proxetil Tablets, and powder. Weigh accurately a solution: both spectra exhibit similar intensities of absorp-
portion of the powder, equivalent to about 0.3 g (potency) of tion at the same wavelengths.
Cefpodoxime Proxetil, add 80 mL of a mixture of water, (2) Determine the infrared absorption spectrum of
acetonitrile and acetic acid (100) (99:99:2), agitate for 10 Cefroxadine Hydrate as directed in the potassium bromide
minutes with the aid of ultrasonic waves, and add a mixture disk method under Infrared Spectrophotometry <2.25>, and
of water, acetonitrile and acetic acid (100) (99:99:2) to make compare the spectrum with the Reference Spectrum or the
exactly 100 mL. Filter this solution through a membrane spectrum of Cefroxadine RS: both spectra exhibit similar in-
filter with a pore size not exceeding 0.45 mm. Discard the tensities of absorption at the same wave numbers.
first 10 mL of the filtrate, pipet 10 mL of the subsequent fil- (3) Determine the 1H spectrum of a solution of Cefroxa-
trate, add exactly 6 mL of the internal standard solution, dine Hydrate in deuterated formic acid for nuclear magnetic
then, add a mixture of water, acetonitrile and acetic acid resonance spectroscopy (1 in 10) as directed under Nuclear
(100) (99:99:2) to make 50 mL, and use this solution as the Magnetic Resonance Spectroscopy <2.21>, using tetramethyl-
sample solution. Separately, weigh accurately an amount of silane for nuclear magnetic resonance spectroscopy as an
Cefpodoxime Proxetil RS, equivalent to about 60 mg (po- internal reference compound: it exhibits three sharp single
tency), dissolve in 60 mL of a mixture of water, acetonitrile signals, A, B and C, at around d 2.8 ppm, at around d 4.1
and acetic acid (100) (99:99:2), add exactly 12 mL of the in- ppm and at around d 6.3 ppm. The ratio of the integrated
ternal standard solution, then add a mixture of water, aceto- intensity of each signal, A:B:C, is about 4:3:1.
nitrile and acetic acid (100) (99:99:2) to make 100 mL, and
D : ＋95 – ＋1089 (0.1 g calcu-
use this solution as the standard solution. Then, proceed as
lated on the anhydrous basis, diluted acetic acid (100) (3 in
directed in the Assay under Cefpodoxime Proxetil.
25), 100 mL, 100 mm).
Purity (1) Heavy metals <1.07>—Weigh 1.0 g of Cefroxa-
＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 5
dine Hydrate in a porcelain crucible, add 10 mL of a solu-
MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS tion of magnesium nitrate hexahydrate in ethanol (95) (1 in
taken 10), mix, burn the ethanol, and carbonize by gently heating.
After cooling, add 2 mL of nitric acid, heat carefully, and
incinerate by ignition at 500 – 6009C. If a carbonized sub-
stance still remains, moisten it with a small amount of nitric
acid (100) (99:99:2) to make 100 mL.
acid, and incinerate again by ignition. After cooling, add 6
Containers and storage Containers—Tight containers. mL of hydrochloric acid, and evaporate on a water bath to
dryness. Moisten the residue with 3 drops of hydrochloric
acid, and add 10 mL of hot water to dissolve the residue by
Cefroxadine Hydrate heating on a water bath. After cooling, adjust the pH be-
tween 3 and 4 with ammonia TS, add 2 mL of dilute acetic
セフロキサジン水和物 acid, filter if necessary, transfer to a Nessler tube, wash the
crucible with 10 mL of water, and add the washing and
water to the tube to make 50 mL. Perform the test with this
solution. Prepare the control solution as follows: Put 2.0 mL
of Standard Lead Solution and 10 mL of a solution of mag-
nesium nitrate hexahydrate in ethanol (95) (1 in 10) in a por-
celain crucible, and proceed as directed for the preparation
of the test solution (not more than 20 ppm).
C16H19N3O5S.2H2O: 401.43
(2) Related substances—Dissolve 10 mg of Cefroxadine
(6R,7R)-7-[(2R)-2-Amino-2-cyclohexa-1,4-
Hydrate in 100 mL of the mobile phase, and use this solution
dienylacetylamino]-3-methoxy-8-oxo-5-thia-1-
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid dihydrate
[51762-05-1, anhydride]
Cefroxadine Hydrate contains not less than 930 mg
peak area by the automatic integration method: the areas of
Cefroxadine Hydrate is expressed as mass (potency) of
the peaks, having the relative retention times of about 0.07,
cefroxadine (C16H19N3O5S: 365.40).
about 0.6 and about 0.8 to cefroxadine obtained from the
Description Cefroxadine Hydrate occurs as pale yellowish sample solution are not larger than 2 times, 4 times and 1
white to light yellow, crystalline particles or powder. time the peak area of cefroxadine obtained from the stand-
It is very soluble in formic acid, slightly soluble in water ard solution, respectively, and any peak area other than
and in methanol, and very slightly soluble in acetonitrile and cefroxadine and other than the peaks mentioned above is not
in ethanol (95). larger than 1/2 times the peak area of cefroxadine from the
It dissolves in 0.001 mol/L hydrochloric acid TS and in standard solution, and the total area of the peaks other than
dilute acetic acid. cefroxadine is not larger than 6 times the peak area of
cefroxadine from the standard solution.
solution of Cefroxadine Hydrate in 0.001 mol/L hydrochlo-
ric acid TS (1 in 50,000) as directed under Ultraviolet-visible
length: 254 nm).
the Reference Spectrum or the spectrum of a solution of
JP XVII Official Monographs / Cefroxadine for Syrup 649
gel for liquid chromatography (5 mm in particle diameter). with 10 mL of the standard solution under the above operat-
Column temperature: A constant temperature of about ing conditions, the relative standard deviation of the ratios
259 C. of the peak areas of cefroxadine to that of the internal stand-
Mobile phase: Dissolve 1.4 g of sodium perchlorate in ard is not more than 1.0z.
1000 mL of a mixture of water and acetonitrile (489:11).
Flow rate: Adjust so that the retention time of cefroxadine
retention time of cefroxadine. Cefroxadine for Syrup
シロップ用セフロキサジン
actly 20 mL. Confirm that the peak area of cefroxadine ob- Cefroxadine for Syrup is a preparation for syrup,
tained with 40 mL of this solution is equivalent to 7 to 13z which is suspended before use.
of that obtained with 40 mL of the standard solution. It contains not less than 93.0z and not more
System performance: Dissolve 3 mg of Cefroxadine Hy- than 107.0z of the labeled potency of cefroxadine
drate and 15 mg of orcin in 100 mL of the mobile phase. (C16H19N3O5S: 365.40).
the above operating conditions, orcin and cefroxadine are
tions for Syrups, with Cefroxadine Hydrate.
being not less than 3. Identification Powder Cefroxadine for Syrup, if necessary.
System repeatability: When the test is repeated 6 times To a portion of the powder, equivalent to 2 mg (potency) of
with 40 mL of the standard solution under the above operat- Cefroxadine Hydrate, add 100 mL of 0.001 mol/L hydro-
ing conditions, the relative standard deviation of the peak chloric acid TS, shake well, and filter. Determine the absorp-
area of cefroxadine is not more than 2.0z. tion spectrum of this filtrate as directed under Ultraviolet-
Assay Weigh accurately an amount of Cefroxadine Hy-
drate and Cefroxadine RS, equivalent to about 50 mg (po-
tency), dissolve each in a suitable amount of a mixture of Uniformity of dosage units <6.02> Perform the test accord-
dilute acetic acid and phosphoric acid (500:1), add exactly 5 ing to the following method: Cefroxadine for Syrup in
mL of the internal standard solution and a mixture of dilute single-dose packages meet the requirement of the Content
acetic acid and phosphoric acid (500:1) to make 200 mL, and uniformity test.
use these solutions as the sample solution and standard solu- Take out the total contents of 1 package of Cefroxadine
tion. Perform the test with 10 mL each of the sample solution for Syrup, add 4V/5 mL of a mixture of dilute acetic acid
and standard solution as directed under Liquid Chromatog- and phosphoric acid (500:1), shake well for 15 minutes, add
raphy <2.01> according to the following conditions, and cal- exactly 5 mL of the internal standard solution per 50 mg (po-
culate the ratios, QT and QS, of the peak area of cefroxadine tency) of Cefroxadine Hydrate, and add a mixture of dilute
to that of the internal standard. acetic acid and phosphoric acid (500:1) to make V mL so
that each mL contains about 0.25 mg (potency) of Cefroxa-
Amount [ mg (potency)] of cefroxadine (C16H19N3O5S)
dine Hydrate. Filter this solution through a membrane filter
＝ MS × QT/QS × 1000
with pore size of not exceeding 0.45 mm, and use the filtrate
MS: Amount [mg (potency)] of Cefroxadine RS taken as the sample solution. Separately, weigh accurately an
amount of Cefroxadine RS, equivalent to about 50 mg (po-
Internal standard solution—Dissolve 1.6 g of vanillin in 5
tency), dissolve in a mixture of dilute acetic acid and phos-
mL of methanol, and add a mixture of dilute acetic acid and
phoric acid (500:1), add exactly 5 mL of the internal stand-
phosphoric acid (500:1) to make 100 mL.
ard solution, add a mixture of dilute acetic acid and phos-
phoric acid (500:1) to make 200 mL, and use this solution as
the standard solution. Then, proceed as directed in the Assay
length: 254 nm).
under Cefroxadine Hydrate.
ter and 10 cm in length, packed with octadecylsilanized silica Amount [mg (potency)] of cefroxadine (C16H19N3O5S)
gel for liquid chromatography (5 mm in particle diameter). ＝ MS × QT/QS × V/200
MS: Amount [mg (potency)] of Cefroxadine RS taken
259 C.
Mobile phase: A mixture of a solution of ammonium sul- Internal standard solution—Dissolve 1.6 g of vanillin in 5
fate (1 in 50) and acetonitrile (97:3). mL of methanol, and add a mixture of dilute acetic acid and
Flow rate: Adjust so that the retention time of cefroxadine phosphoric acid (500:1) to make 100 mL.
in 15 minutes of Cefroxadine for Syrup is not less than 85z.
ditions, cefroxadine and the internal standard are eluted in
Cefroxadine for Syrup, equivalent to about 0.1 g (potency)
less than 1.5.
of Cefroxadine Hydrate, withdraw not less than 10 mL of
the medium at the specified minute after starting the test,
650 Cefsulodin Sodium / Official Monographs JP XVII
and filter through a membrane filter with a pore size not Cefsulodin Sodium is expressed as mass (potency) of
exceeding 0.8 mm. Discard the first 5 mL of the filtrate, pipet cefsulodin (C22H20N4O8S2: 532.55).
4 mL of the subsequent filtrate, add 0.1 mol/L hydrochloric
Description Cefsulodin Sodium occurs as white to light yel-
low, crystals or crystalline powder.
It is freely soluble in water and in formamide, slightly
Cefroxadine RS, equivalent to about 22 mg (potency), and
soluble in methanol, and very slightly soluble in ethanol (95).
dissolve in 0.1 mol/L hydrochloric acid TS to make exactly
It is hygroscopic.
100 mL. Pipet 5 mL of this solution, add 10 mL of water,
add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, Identification (1) Determine the absorption spectrum of a
and use this solution as the standard solution. Perform the solution of Cefsulodin Sodium (1 in 50,000) as directed
test with the sample solution and standard solution as di- under Ultraviolet-visible Spectrophotometry <2.24>, and
rected under Ultraviolet-visible Spectrophotometry <2.24>, compare the spectrum with the Reference Spectrum or the
and determine the absorbances, AT and AS, at 267 nm. spectrum of a solution of Cefsulodin Sodium RS prepared in
the same manner as sample solution: both spectra exhibit
of cefroxadine (C16H19N3O5S)
＝ MS/MT × AT/AS × 1/C × 450
sulodin Sodium as directed in the potassium bromide disk
MS: Amount [mg (potency)] of Cefroxadine RS taken method under Infrared Spectrophotometry <2.25>, and com-
MT: Amount (g) of Cefroxadine for Syrup taken pare the spectrum with the Reference Spectrum or the spec-
C: Labeled amount [mg (potency)] of cefroxadine trum of Cefsulodin Sodium RS: both spectra exhibit similar
(C16H19N3O5S) in 1 g intensities of absorption at the same wave numbers.
(3) Determine the 1H spectrum of a solution of Cef-
Assay Powder Cefroxadine for Syrup, if necessary, weigh
sulodin Sodium in heavy water for nuclear magnetic
accurately a portion of the powder, equivalent to about 50
mg (potency) of Cefroxadine Hydrate, add 160 mL of a mix-
ture of dilute acetic acid and phosphoric acid (500:1), shake
well for 15 minutes, add exactly 5 mL of the internal stand-
ard solution, and add a mixture of dilute acetic acid and
hibits a multiple signal A between d 7.3 ppm and d 7.7 ppm,
phosphoric acid (500:1) to make 200 mL. Filter this solution
and double signals, B and C, at around d 8.4 ppm and at
around d 9.1 ppm, respectively. The ratio of integrated in-
0.45 mm, and use the filtrate as the sample solution. Sepa-
tensity of these signals, A:B:C, is about 5:2:2.
rately, weigh accurately an amount of Cefroxadine RS,
(4) Cefsulodin Sodium responds to the Qualitative Tests
equivalent to about 50 mg (potency), dissolve in a mixture of
<1.09> (1) for sodium salt.
dilute acetic acid and phosphoric acid (500:1), add exactly 5
mL of the internal standard solution, add a mixture of dilute Optical rotation <2.49> [a]20
D : ＋16.5 – ＋20.09(0.1 g calcu-
acetic acid and phosphoric acid (500:1) to make 200 mL, and lated on the anhydrous basis, water, 10 mL, 100 mm).
use this solution as the standard solution. Proceed as di-
pH <2.54> Dissolve 1.0 g of Cefsulodin Sodium in 10 mL
rected in the Assay under Cefroxadine Hydrate.
of water: the pH of the solution is not less than 3.3 and not
Amount [mg (potency)] of cefroxadine (C16H19N3O5S) more than 4.8.
＝ M S × QT / QS
Purity (1) Clarity of solution—Dissolve 1.0 g of Cef-
MS: Amount [mg (potency)] of Cefroxadine RS taken sulodin Sodium in 10 mL of water: the solution is clear.
(2) Heavy metals <1.07>—To 1.0 g of Cefsulodin Sodium
Internal standard solution—Dissolve 1.6 g of vanillin in 5
add 10 mL of a solution of magnesium nitrate hexahydrate
mL of methanol, and add a mixture of dilute acetic acid and
in ethanol (95) (1 in 5), mix, fire the ethanol to burn, then
phosphoric acid (500:1) to make 100 mL.
heat gradually to carbonize. After cooling, add 2 mL of
Containers and storage Containers—Tight containers. nitric acid, heat carefully, then heat at 500 – 6009C to incin-
erate. If a carbonized residue still retains, add a little amount
of nitric acid, and heat again to incinerate. After cooling,
Cefsulodin Sodium add 6 mL of hydrochloric acid to the residue, heat to dryness
on a water bath, then moisten the residue with 3 drops of hy-
セフスロジンナトリウム drochloric acid, add 10 mL of hot water, and heat on a water
bath to dissolve. Add ammonia TS dropwise to adjust to pH
3 – 4, and add 2 mL of dilute acetic acid. If necessary, filter,
wash the crucible and residue on the filter with 10 mL of
water, transfer the filtrate and washings into a Nessler tube,
add water to make 50 mL, and use this solution as the test
solution. Prepare the control solution as follows: To 2.0 mL
of Standard Lead Solution add 10 mL of a solution of mag-
C22H19N4NaO8S2: 554.53
nesium nitrate hexahydrate in ethanol (95) (1 in 5), fire the
Monosodium (6R,7R)-3-(4-carbamoylpyridinium-1-
ethanol to burn. After cooling, add 2 mL of nitric acid, heat
ylmethyl)-8-oxo-7-[(2R)-2-phenyl-2-sulfonatoacetylamino]-
carefully, then heat at 500 – 6009C. After cooling, add 6 mL
5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate
of hydrochloric acid, then proceed in the same manner as for
[52152-93-9]
the preparation of the test solution (not more than 20 ppm).
Cefsulodin Sodium contains not less than 900 mg
of Cefsulodin Sodium according to Method 3, using a solu-
tion of magnesium nitrate hexahydrate in ethanol (95) (1 in
JP XVII Official Monographs / Cefsulodin Sodium 651
5) and 15 mL of dilute hydrochloric acid instead of a solu- System repeatability: When the test is repeated 5 times
tion of magnesium nitrate hexahydrate in ethanol (95) (1 in with 10 mL of the standard solution under the above operat-
50) and 3 mL of hydrochloric acid, and perform the test (not ing conditions, the relative standard deviation of the peak
more than 2 ppm). areas of cefsulodin is not more than 1.0z.
(4) Related substances—Weigh accurately 0.10 g of Cef-
sulodin Sodium, dissolve in water to make exactly 50 mL,
tion, direct titration, avoiding moisture absorption of the
sample, using a mixture of formamide for water determina-
weigh accurately about 20 mg of isonicotinic acid amide and
tion and methanol for water determination (2:1) instead of
about 20 mg of Cefsulodin Sodium RS (separately determine
methanol for water determination).
the water <2.48> in the same manner as Cefsulodin Sodium),
and dissolve in water to make exactly 100 mL. Pipet 10 mL Assay Weigh accurately an amount of Cefsulodin Sodium
of this solution, add water to make exactly 100 mL, and use and Cefsulodin Sodium RS, equivalent to about 0.1 g (po-
this solution as the standard solution. Perform the test with tency), dissolve each in water to make exactly 50 mL, and
exactly 10 mL each of the sample solution and standard solu- use these solutions as the sample solution and standard solu-
tion as directed under Liquid Chromatography <2.01> ac- tion. Perform the test with exactly 10 mL each of the sample
cording to the following conditions, and determine the areas solution and standard solution as directed under Liquid
of each peak by the automatic integration method. Calculate Chromatography <2.01> according to the following condi-
the amount of the related substances by the following for- tions, and determine the peak areas, AT and AS, of cef-
mula: the amount of isonicotinic acid amide is not more than sulodin in each solution.
1.0z, and the total amount of other related substances is
Amount [ mg (potency)] of cefsulodin (C22H20N4O8S2)
not more than 1.2z.
＝ MS × AT/AS × 1000
Amount (z) of isonicotinic acid amide
MS: Amount [mg (potency)] of Cefsulodin Sodium RS
＝ A/BI × MI/MT × 5
taken
Total amount (z) of the other related substances
＝ B/BS × MS/MT × 5
A: Peak area of isonicotinic acid amide from the sample length: 254 nm).
solution Column: A stainless steel column 4 mm in inside diameter
B: Total peak area other than cefsulodin and other than and 15 cm in length, packed with octadecylsilanized silica gel
isonicotinic acid amide from the sample solution for liquid chromatography (5 mm in particle diameter).
BI: Peak area of isonicotinic acid amide from the standard Column temperature: A constant temperature of about
solution 259C.
BS: Peak area of cefsulodin from the standard solution Mobile phase: A mixture of a solution of ammonium sul-
MT: Amount (g) of Cefsulodim Sodium taken fate (1 in 100) and acetonitrile (97:3).
MS: Amount (g) of Cefsulodin Sodium RS taken Flow rate: Adjust so that the retention time of cefsulodin
MI: Amount (g) of isonicotinic acid amide taken is about 9 minutes.
System performance: Dissolve 40 mg of isonicotinic acid
amide in 25 mL of the standard solution. When the proce-
length: 254 nm).
dure is run with 10 mL of this solution under the above oper-
ating conditions, isonicotinic acid amide and cefsulodin are
259 C.
Mobile phase A: A mixture of a solution of ammonium
sulfate (1 in 100) and acetonitrile (97:3).
areas of cefsulodin is not more than 1.0z.
Mobile phase B: A mixture of a solution of ammonium
sulfate (1 in 100) and acetonitrile (23:2). Containers and storage Containers—Hermetic containers.
Flowing of mobile phase: Change the mobile phase A to B
at 14 minutes after the injection of sample.
Flow rate: Adjust so that the retention time of cefsulodin
is about 9 minutes.
retention time of cefsulodin.
solution, add water to make exactly 10 mL. Confirm that the
peak areas of isonicotinic acid amide and cefsulodin ob-
tained from 10 mL of this solution are equivalent to 7 to 13z
of those of isonicotinic acid amide and cefsulodin obtained
ditions, isonicotinic acid amide and cefsulodin are eluted in
less than 5.
652 Ceftazidime Hydrate / Official Monographs JP XVII
(2) Heavy metals <1.07>—Proceed with 1.0 g of Ceftazi-
Ceftazidime Hydrate dime Hydrate according to Method 2, and perform the test.
セフタジジム水和物 Solution (not more than 20 ppm).
(3) Related substances (i) Trityl-t-butyl substance and
t-butyl substance—Dissolve 0.10 g of Ceftazidime Hydrate
in 2 mL of diluted disodium hydrogen phosphate TS (1 in 3),
the sample solution, add diluted disodium hydrogen-
phosphate TS (1 in 3) to make exactly 100 mL, and use this
C22H22N6O7S2.5H2O: 636.65
(6R,7R)-7-[(Z )-2-(2-Aminothiazol-4-yl)-2-(1-carboxy-
1-methylethoxyimino)acetylamino]-3-(pyridinium-1-
thin-layer chromatography. Develop with a mixture of acetic
ylmethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-
acid (100), n-butyl acetate, acetate buffer solution (pH 4.5)
carboxylate pentahydrate
and 1-butanol (16:16:13:3) to a distance of about 12 cm, and
[78439-06-2]
wavelength: 254 nm): the spots which appear upper in posi-
Ceftazidime Hydrate contains not less than 950 mg
tion than the principal spot obtained from the sample solu-
tion are not more intense than the spot obtained from the
standard solution.
Ceftazidime Hydrate is expressed as mass (potency) of
(ii) Other related substances—Dissolve 20 mg of Ceftazi-
ceftazidime (C22H22N6O7S2: 546.58).
dime Hydrate in 10 mL of the mobile phase, and use this
Description Ceftazidime Hydrate occurs as a white to light solution as the sample solution. Pipet 1 mL of the sample so-
yellowish white crystalline powder. lution, add the mobile phase to make exactly 200 mL, and
It is slightly soluble in water, and very slightly soluble in use this solution as the standard solution. Perform the test
acetonitrile and in ethanol (95). with exactly 5 mL each of the sample solution and standard
solution of Ceftazidime Hydrate in phosphate buffer solu-
tion (pH 6.0) (1 in 100,000) as directed under Ultraviolet-
the peak other than ceftazidime obtained from the sample
solution is not larger than that of ceftazidime obtained from
with the Reference Spectrum or the spectrum of a solution of
the standard solution, and the total of peak areas other than
Ceftazidime RS prepared in the same manner as the sample
ceftazidime from the sample solution is not larger than 5
solution: both spectra exhibit similar intensities of absorp-
times the peak area of ceftazidime from the standard solu-
tion at the same wavelengths.
tion.
Ceftazidime Hydrate as directed in the potassium bromide
length: 254 nm).
spectrum of Ceftazidime RS: both spectra exhibit similar in-
(3) To 0.05 g of Ceftazidime Hydrate add 5 mg of dried
sodium carbonate, and add 0.5 mL of heavy water for
259C.
nuclear magnetic resonance spectroscopy to dissolve. Deter-
Mobile phase: Dissolve 5.0 g of ammonium dihydrogen-
mine the 1H spectrum of this solution as directed under
phosphate in 750 mL of water, adjust to pH 3.5 with phos-
Nuclear Magnetic Resonance Spectroscopy <2.21>, using so-
phoric acid, and add water to make 870 mL. To this solution
dium 3-trimethylsilylpropanesulfonate for nuclear magnetic
add 130 mL of acetonitrile.
resonance spectroscopy as an internal reference compound:
Flow rate: Adjust so that the retention time of ceftazidime
it exhibits single signals, A and B, at around d 1.5 ppm and
is about 4 minutes.
at around d 6.9 ppm, and a multiple signal C between d 7.9
ppm and d 9.2 ppm. The ratio of integrated intensity of each
retention time of ceftazidime, beginning after the solvent
signal, A:B:C, is about 6:1:5.
peak.
D : －28 – －349(0.5 g calculated System suitability—
on the anhydrous basis, phosphate buffer solution (pH 6.0), Test for required detectability: Pipet 1 mL of the standard
100 mL, 100 mm). solution, add the mobile phase to make exactly 5 mL, and
confirm that the peak area of ceftazidime obtained with 5 mL
pH <2.54> Dissolve 0.5 g of Ceftazidime Hydrate in 100
of this solution is equivalent to 15 to 25z of that obtained
Purity (1) Clarity and color of solution—Dissolve 1.0 g System performance: Dissolve about 10 mg each of
of Ceftazidime Hydrate in 10 mL of a solution obtained by Ceftazidime Hydrate and acetanilide in 20 mL of the mobile
dissolving 5 g of anhydrous disodium hydrogen phosphate phase. When the procedure is run with 5 mL of this solution
and 1 g of potassium dihydrogen phosphate in water to make under the above operating conditions, ceftazidime and
100 mL: the solution is clear, and its absorbance at 420 nm, acetanilide are eluted in this order with the resolution be-
determined as directed under Ultraviolet-visible Spectropho- tween these peaks being not less than 10.
tometry <2.24>, is not more than 0.20. System repeatability: When the test is repeated 6 times
JP XVII Official Monographs / Ceftazidime for Injection 653
with 5 mL of the standard solution under the above operating Amount [mg (potency)] of ceftazidime (C22H22N6O7S2)
conditions, the relative standard deviation of the peak area ＝ MS × QT/QS × 5000
of ceftazidime is not more than 2.0z.
MS: Amount [mg (potency)] of Ceftazidime RS taken
(4) Free pyridine—Weigh accurately about 50 mg of
Ceftazidime Hydrate, dissolve in the mobile phase to make Internal standard solution—A solution of dimedon in 0.05
exactly 10 mL, and use this solution as the sample solution. mol/L phosphate buffer solution (pH 7.0) (11 in 10,000).
Separately, weigh accurately about 0.1 g of pyridine, and Operating conditions—
add the mobile phase to make exactly 100 mL. Pipet 1 mL of Detector: An ultraviolet absorption photometer (wave-
this solution, add the mobile phase to make exactly 100 mL, length: 254 nm).
and use this solution as the standard solution. Perform the Column: A stainless steel column 4.6 mm in inside diame-
test with exactly 10 mL each of the sample solution and ter and 10 cm in length, packed with hexasilanized silica gel
standard solution as directed under Liquid Chromatography for liquid chromatography (5 mm in particle diameter).
<2.01> according to the following conditions, and determine Column temperature: A constant temperature of about
the peak heights, HT and HS, of pyridine in each solution: 259C.
the amount of free pyridine is not more than 0.3z. Mobile phase: Dissolve 4.26 g of anhydrous disodium
Amount (mg) of free pyridine
phosphate in 980 mL of water, and add 20 mL of aceto-
＝ MS × HT/HS × 1/1000
nitrile.
MS: Amount (mg) of pyridine taken Flow rate: Adjust so that the retention time of ceftazidime
is about 4 minutes.
length: 254 nm).
tions, the internal standard and ceftazidime are eluted in this
than 3.
409 C.
phosphate in 500 mL of water, add 300 mL of acetonitrile
the peak area of ceftazidime to that of the internal standard
and water to make 1000 mL, and adjust to pH 7.0 with
ammonia solution (28).
Flow rate: Adjust so that the retention time of pyridine is Containers and storage Containers—Tight containers.
about 4 minutes. Storage—Light-resistant.
System performance: Dissolve 5 mg of Ceftazidime Hy-
drate in 100 mL of a solution of pyridine in the mobile phase Ceftazidime for Injection
(1 in 20,000). When the procedure is run with 10 mL of this
solution under the above operating conditions, ceftazidime 注射用セフタジジム
and pyridine are eluted in this order with the resolution be-
tween these peaks being not less than 9.
Ceftazidime for Injection is a preparation for injec-
tion which is dissolved before use.
than 107.0z of the labeled potency of ceftazidime
height of pyridine is not more than 5.0z.
(C22H22N6O7S2: 546.58).
direct titration).
tions, with Ceftazidime Hydrate.
Assay Weigh accurately an amount of Ceftazidime Hy-
Description Ceftazidime for Injection is a white to pale yel-
drate, equivalent to about 0.1 g (potency), and dissolve in
0.05 mol/L phosphate buffer solution (pH 7.0) to make ex-
actly 100 mL. Pipet 10 mL of this solution, add exactly 5 mL Identification Determine the absorption spectrum of a so-
of the internal standard solution, then add 0.05 mol/L phos- lution of Ceftazidime for Injection (1 in 100,000) in phos-
phate buffer solution (pH 7.0) to make 50 mL, and use this phate buffer solution (pH 6.0) as directed under Ultraviolet-
solution as the sample solution. Separately, weigh accurately visible Spectrophotometry <2.24>: it exhibits a maximum
an amount of Ceftazidime RS, equivalent to about 20 mg between 255 nm and 259 nm.
(potency), dissolve in 0.05 mol/L phosphate buffer solution
pH <2.54> Dissolve an amount of Ceftazidime for Injec-
(pH 7.0) to make exactly 20 mL. Pipet 10 mL of this solu-
tion, equivalent to 1.0 g (potency) of Ceftazidime Hydrate,
in 10 mL of water: the pH of this solution is 5.8 to 7.8.
then add 0.05 mol/L phosphate buffer solution (pH 7.0) to
make 50 mL, and use this solution as the standard solution. Purity Clarity and color of solution—Dissolve 5 g of diso-
Perform the test with 5 mL each of the sample solution and dium hydrogen phosphate and 1 g of potassium dihydrogen
standard solution as directed under Liquid Chromatography phosphate in water to make 100 mL. In 10 mL of this solu-
<2.01> according to the following conditions, and calculate tion dissolve an amount of Ceftazidime for Injection,
the ratios, QT and QS, of the peak area of ceftazidime to that equivalent to 1.0 g (potency) of Ceftazidime Hydrate: the so-
of the internal standard. lution is clear. Also, determine the absorption spectra of this
654 Cefteram Pivoxil / Official Monographs JP XVII
tometry <2.24>: the absorbance at 420 nm is not more than calculated on the anhydrous basis. The potency of
0.3. Cefteram Pivoxil is expressed as mass (potency) of
cefteram (C16H17N9O5S2: 479.49).
Loss on drying <2.41> Not more than 14.0z (0.1 g, in
vacuum not exceeding 0.67 kPa, 609C, 3 hours). Description Cefteram Pivoxil occurs as a white to pale yel-
It is very soluble in acetonitrile, freely soluble in metha-
tency).
nol, in ethanol (95) and in chloroform, and practically in-
Uniformity of dosage units <6.02> It meets the requirement soluble in water.
Foreign insoluble matter <6.06> Perform the test according solution of Cefteram Pivoxil in 0.05 mol/L hydrochloric
to Method 2: it meets the requirement. acid-methanol TS (1 in 100,000) as directed under Ultravio-
let-visible Spectrophotometry <2.24>, and compare the spec-
trum with the Reference Spectrum: both spectra exhibit simi-
ment.
lar intensities of absorption at the same wavelengths.
Sterility <4.06> Perform the test according to the Mem- (2) Determine the infrared absorption spectrum of
brane filter method: it meets the requirement. Cefteram Pivoxil as directed in the potassium bromide disk
than 10 containers of Ceftazidime for Injection. Weigh accu-
rately an amount of Ceftazidime Hydrate, equivalent to
numbers.
about 0.25 g (potency), and dissolve in 0.05 mol/L phos-
(3) Determine the 1H spectrum of a solution of Cefteram
phate buffer solution (pH 7.0) to make exactly 250 mL.
Pivoxil in deuterated chloroform for nuclear magnetic reso-
nance spectroscopy (1 in 10) as directed under Nuclear
nal standard solution, add more 0.05 mol/L phosphate
Magnetic Resonance Spectroscopy <2.21>, using tetramethy-
buffer solution (pH 7.0) to make 50 mL, and use this solu-
lsilane for nuclear magnetic resonance spectroscopy as an
internal reference compound: it exhibits single signals A, B
amount of Ceftazidime RS, equivalent to about 25 mg (po-
and C, at around d 1.2 ppm, at around d 2.5 ppm and at
around d 4.0 ppm, respectively. The ratio of the integrated
(pH 7.0) to make exactly 25 mL. Pipet 10 mL of this solu-
intensity of these signals, A:B:C, is about 3:1:1.
then add 0.05 mol/L phosphate buffer solution (pH 7.0) to Optical rotation <2.49> [a]20
D : ＋35 – ＋439(0.4 g calculated
make 50 mL, and use this solution as the standard solution. on the anhydrous basis, methanol, 20 mL, 100 mm).
Then, proceed as directed in the Assay under Ceftazidime
Hydrate.
Cefteram Pivoxil according to Method 2, and perform the
Amount [mg (potency)] of ceftazidime (C22H22N6O7S2) test. Prepare the control solution with 2.0 mL of Standard
＝ MS × QT/QS × 10 Lead Solution (not more than 20 ppm).
(2) Related substances—Dissolve 50 mg of Cefteram
MS: Amount [mg(potency)] of Ceftazidime RS taken
Pivoxil in 50 mL of the mobile phase, and use this solution
Internal standard solution—A solution of dimedon in 0.05 as the sample solution. Pipet 1 mL of the sample solution,
mol/L phosphate buffer solution (pH 7.0) (11 in 10,000). add the mobile phase to make exactly 50 mL, and use this so-
the automatic integration method: the each area of the
Cefteram Pivoxil peaks, having the relative retention time of about 0.2 and
about 0.9 to cefteram pivoxil, obtained from the sample so-
セフテラムピボキシル
lution is not larger than 1/2 times and 1.25 times the peak
area of cefteram pivoxil obtained from the standard solu-
tion, respectively, the area of the peak other than cefteram
pivoxil and the peaks mentioned above is not larger than 1/4
times the peak area of cefteram pivoxil from the standard so-
lution, and the total area of the peaks other than cefteram
pivoxil is not larger than 2.75 times the peak area of cefter-
am pivoxil from the standard solution. For the area of the
peak, having the relative retention time of about 0.1 to
cefteram pivoxil, multiply the relative response factor, 0.74.
C22H27N9O7S2: 593.64 Operating conditions—
2,2-Dimethylpropanoyloxymethyl (6R,7R)-7-[(Z )-2- Detector, column, column temperature, mobile phase, and
(2-aminothiazol-4-yl)-2-(methoxyimino)acetylamino]-3- flow rate: Proceed as directed in the operating conditions in
(5-methyl-2H-tetrazol-2-ylmethyl)-8-oxo-5-thia-1- the Assay.
azabicyclo[4.2.0]oct-2-ene-2-carboxylate Time span of measurement: About 2 times as long as the
[82547-58-8, Cefteram] retention time of cefteram pivoxil.
Cefteram Pivoxil contains not less than 743 mg (po- Test for required detectability: To exactly 1 mL of the
tency) and not more than 824 mg (potency) per mg, standard solution add the mobile phase to make exactly 10
JP XVII Official Monographs / Cefteram Pivoxil Fine Granules 655
mL. Confirm that the peak area of cefteram pivoxil obtained

from 10 mL of this solution is equivalent to 7 to 13z of that Cefteram Pivoxil Fine Granules
obtained from 10 mL of the standard solution.
System performance: When the procedure is run with 10 セフテラムピボキシル細粒
Cefteram Pivoxil Fine Granules contain not less
factor of the peak of cefteram pivoxil are not less than 5000
potency of cefteram (C16H17N9O5S2: 479.49).
with 10 mL of the standard solution under the above operat- Method of preparation Prepare as directed under Gran-
ing conditions, the relative standard deviation of the peak ules, with Cefteram Pivoxil.
area of cefteram pivoxil is not more than 3.0z.
Identification Powder Cefteram Pivoxil Fine Granules. To
Water <2.48> Not more than 3.0z (0.3 g, coulometric a portion of the powder, equivalent to 0.1 g (potency) of
titration). Cefteram Pivoxil, add 20 mL of methanol, shake well, and
filter. To 1 mL of the filtrate add 0.05 mol/L hydrochloric
Assay Weigh accurately an amount of Cefteram Pivoxil
acid-methanol TS to make 500 mL, and determine the ab-
and Cefteram Pivoxil Mesitylene Sulfonate RS, equivalent to
sorption spectrum as directed under Ultraviolet-visible Spec-
about 40 mg (potency), dissolve each in 20 mL of diluted
trophotometry <2.24>: it exhibits a maximum between 262
acetonitrile (1 in 2), add exactly 5 mL of the internal stand-
nm and 266 nm.
ard solution and diluted acetonitrile (1 in 2) to make 50 mL,
and use these solutions as the sample solution and standard Purity Related substances—Powder Cefteram Pivoxil Fine
solution. Perform the test with 10 mL each of the sample so- Granules, if necessary. To a portion, equivalent to 0.1 g (po-
lution and standard solution as directed under Liquid Chro- tency) of Cefteram Pivoxil, add diluted acetonitrile (1 in 2)
matography <2.01> according to the following conditions, to make 100 mL, disperse the particle with the aid of ultra-
and calculate the ratios, QT and QS, of the peak area of sonic waves, then filter, and use the filtrate as the sample so-
cefteram pivoxil to that of the internal standard. lution. Pipet 1 mL of the sample solution, add the mobile
Amount [ mg (potency)] of cefteram (C16H17N9O5S2)
＝ MS × QT/QS × 1000
MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity- under Liquid Chromatography <2.01> according to the fol-
lene Sulfonate RS taken lowing conditions, and determine each peak area by the au-
tomatic integration method: the area of the peak, having the
relative retention time of about 0.9 to cefteram pivoxil ob-
droxybenzoate in diluted acetonitrile (1 in 2) (1 in 1000).
tained from the sample solution, is not larger than 1.75 times
the peak area of cefteram pivoxil obtained from the standard
solution, the area of the peak, having the relative retention
length: 254 nm).
time of about 0.1 from the sample solution, is not larger
than 17/25 times the peak area of cefteram pivoxil from the
cefteram pivoxil from the sample solution is not larger than
3.7 times the peak area of cefteram pivoxil from the stand-
259 C.
ard solution. For the area of the peak, having the relative
Mobile phase: To 100 mL of acetic acid-sodium acetate
retention time of about 0.1 to cefteram pivoxil, multiply the
buffer solution (pH 5.0) add 375 mL of acetonitrile and
relative response factor, 0.74.
water to make 1000 mL.
Flow rate: Adjust so that the retention time of cefteram
Proceed as directed in the operating conditions in tne
pivoxil is about 14 minutes.
Purity (2) under Cefteram Pivoxil.
(2) under Cefteram Pivoxil.
ditions, the internal standard and cefteram pivoxil are eluted
in this order with the resolution between these peaks being Water <2.48> Not more than 0.3z (0.1 g (potency), coulo-
not less than 3. metric titration).
Uniformity of dosage units <6.02> The Granules in single-
test.
of the peak area of cefteram pivoxil to that of the internal
standard is not more than 1.0z. Dissolution Being specified separately when the drug is
Storage—In a cold place. Assay Powder Cefteram Pivoxil Fine Granules, if neces-
sary. Weigh accurately an amount of the powder, equivalent
to about 0.3 g (potency) of Ceteram Pivoxil, add exactly 30
mL of the internal standard solution and diluted acetonitrile
(1 in 2) to make 300 mL. Disperse the particle with the aid of
ultrasonic waves, then filter, and use the filtrate as the sam-
ple solution. Separately, weigh accurately an amount of
Cefteram Pivoxil Mesitylene Sulfonate RS, equivalent to
656 Cefteram Pivoxil Tablets / Official Monographs JP XVII
about 50 mg (potency), dissolve in 20 mL of diluted aceto- retention time of about 0.1 to cefteram pivoxil, multiply the
nitrile (1 in 2), add exactly 5 mL of the internal standard so- relative response factor, 0.74.
lution and diluted acetonitrile (1 in 2) to make 50 mL, and Operating conditions—
use this solution as the standard solution. Perform the test Proceed as directed in the operating conditions in the
with 10 mL each of the sample solution and standard solution Purity (2) under Cefteram Pivoxil.
as directed under Liquid Chromatography <2.01> according System suitability—
to the following conditions, and calculate the ratios, QT and Proceed as directed in the system suitability in the Purity
QS, of the peak area of cefteram pivoxil to that of the inter- (2) under Cefteram Pivoxil.
nal standard.
Water <2.48> Not more than 4.0z (a quantity equivalent
Amount [mg (potency)] of cefteram (C16H17N9O5S2) to 0.2 g (potency) of powdered Cefteram Pivoxil Tablets,
＝ MS × QT/QS × 6 volumetric titration, direct titration).
MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity- Uniformity of dosage units <6.02> Perform the Mass varia-
lene Sulfonate RS taken tion test, or the Content uniformity test according to the fol-
To 1 tablet of Cefteram Pivoxil Tablets add exactly 5 mL
droxybenzoate in diluted acetonitrile (1:2) (1 in 1000).
of the internal standard solution per 50 mg (potency) of
Cefteram Pivoxil, and add diluted acetonitrile (1 in 2) to
make V mL so that each mL contains about 1 mg (potency)
Assay under Cefteram Pivoxil.
of Cefteram Pivoxil. Disperse this solution with ultrasonic
waves, filter through a membrane filter with pore size of not
Proceed as directed in the system suitability in the Assay
exceeding 0.45 mm, discard the first 10 mL of the filtrate,
under Cefteram Pivoxil.
and use the subsequent filtrate as the sample solution. Sepa-
Containers and storage Containers—Tight containers. rately, weigh accurately an amount of Cefteram Pivoxil
Mesitylene Sulfonate RS, equivalent to about 50 mg (po-
tency), dissolve in 20 mL of diluted acetonitrile (1 in 2), add
Cefteram Pivoxil Tablets exactly 5 mL of the internal standard solution, add diluted
acetonitrile (1 in 2) to make 50 mL, and use this solution as
セフテラムピボキシル錠 the standard solution. Then, proceed as directed in the Assay
under Cefteram Pivoxil.
Cefteram Pivoxil Tablets contain not less than Amount [mg (potency)] of cefteram (C16H17N9O5S2)
90.0z and not more than 110.0z of the labeled ＝ MS × QT/QS × V/50
potency of cefteram (C16H17N9O5S2: 479.49).
MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity-
Method of preparation Prepare as directed under Tablets, lene Sulfonate RS taken
with Cefteram Pivoxil.
Identification To a quantity of powdered Cefteram Pivoxil droxybenzoate in diluted acetonitrile (1 in 2) (1 in 1000).
Tablets, equivalent to 0.1 g (potency) of Cefteram Pivoxil,
add 20 mL of methanol, shake well, and filter. To 1 mL of
the filtrate add 0.05 mol/L hydrochloric acid-methanol TS
to make 500 mL. Determine the absorption spectrum of this
in 30 minutes of Cefteram Pivoxil Tablets is not less than
75z.
tometry <2.24>: it exhibits a maximum between 262 nm and
Start the test with 1 tablet of Cefteram Pivoxil Tablets,
266 nm.
Purity Related substances—To a quantity of powdered minute after starting the test, and filter through a membrane
Cefteram Pivoxil Tablets, equivalent to 0.1 g (potency) of filter with a pore size not exceeding 0.45 mm. Discard the
Cefteram Pivoxil, add diluted acetonitrile (1 in 2) to make first 10 mL of the filtrate, pipet V mL of the subsequent fil-
100 mL. Disperse this solution with ultrasonic waves, filter, trate, add water to make exactly V? mL so that each mL con-
and use the filtrate as the sample solution. Pipet 1 mL of the tains about 22 mg (potency) of Cefteram Pivoxil, and use this
sample solution, add the mobile phase to make exactly 50 solution as the sample solution. Separately, weigh accurately
mL and use this solution as the standard solution. Perform an amount of Cefteram Pivoxil Mesitylene Sulfonate RS,
the test with exactly 10 mL each of the sample solution and equivalent to about 22 mg (potency), and dissolve in 20 mL
standard solution as directed under Liquid Chromatography of methanol, and add water to make exactly 50 mL. Pipet 5
<2.01> according to the following conditions. Determine each mL of this solution, add water to make exactly 100 mL, and
peak area of both solutions by the automatic integration use this solution as the standard solution. Perform the test
method: the area of the peak, having the relative retention with the sample solution and standard solution as directed
time of about 0.9 to cefteram pivoxil, obtained from the under Ultraviolet-visible Spectrophotometry <2.24>, using
sample solution is not larger than 1.75 times the peak area of water as the blank, and determine the absorbances, AT and
cefteram pivoxil obtained from the standard solution, and AS, at 300 nm.
the area of the peak, having the relative retention time of
about 0.1 from the sample solution is not larger than 17/25
of cefteram (C16H17N9O5S2)
times the peak area of cefteram pivoxil from the standard so-
＝ MS × AT/AS × V?/V × 1/C × 90
lution. Furthermore, the total area of the peaks other than
cefteram pivoxil from the sample solution is not larger than MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity-
3.7 times the peak area of cefteram pivoxil from the stand- lene Sulfonate RS taken
ard solution. For the area of the peak, having the relative C: Labeled amount [mg (potency)] of cefteram
JP XVII Official Monographs / Ceftibuten Hydrate 657
(C16H17N9O5S2) in 1 tablet spectrum with the Reference Spectrum: both spectra exhibit
Assay To a number of tablet of Cefteram Pivoxil Tablets,
(3) Determine the 1H spectrum of a solution of Ceftibut-
equivalent to about 1.0 g (potency) of Cefteram Pivoxil, add
en Hydrate in deuterated dimethyl sulfoxide for nuclear
120 mL of diluted acetonitrile (1 in 2), disperse with ultra-
magnetic resonance spectroscopy (1 in 30), using tetra-
sonic waves, and add diluted acetonitrile (1 in 2) to make ex-
methylsilane for nuclear magnetic resonance spectroscopy as
actly 200 mL. Centrifuge this solution, pipet 10 mL of the
an internal reference compound, as directed under Nuclear
supernatant liquid, add exactly 5 mL of the internal standard
Magnetic Resonance Spectroscopy <2.21>: it exhibits double
solution, add diluted acetonitrile (1 in 2) to make 50 mL,
signals A and B, at around d 3.2 ppm and at around d 5.1
filter through a membrane filter with pore size not exceeding
ppm, a quartet signal C, at around d 5.8 ppm, and a single
0.45 mm, discard the first 3 mL of the filtrate, and use the
signal D, at around d 6.3 ppm. The ratio of integrated inten-
sity of each signal except the signal at around d 3.2 ppm,
accurately an amount of Cefteram Pivoxil Mesitylene Sul-
B:C:D is about 1:1:1.
fonate RS, equivalent to about 50 mg (potency), dissolve in
20 mL of diluted acetonitrile (1 in 2), add exactly 5 mL of Optical rotation <2.49> [a]20D : ＋135 – ＋1559(0.3 g calcu-
the internal standard solution, add diluted acetonitrile (1 in lated on the anhydrous basis, 0.1 mol/L phosphate buffer
2) to make 50 mL, and use this solution as the standard solu- solution for antibiotics, pH 8.0, 50 mL, 100 mm).
tion. Then, proceed as directed in the Assay under Cefteram
Pivoxil.
Ceftibuten Hydrate according to Method 2, and perform the
Amount [mg (potency)] of cefteram (C16H17N9O5S2) test. Prepare the control solution with 2.0 mL of Standard
＝ MS × QT/QS × 20 Lead Solution (not more than 10 ppm).
(2) Related substances—(i) Keep the sample solution
MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity-
and the standard solution at not exceeding 59C and use
lene Sulfonate RS taken
within 2 hours after preparation. Dissolve 25 mg of Cefti-
Internal standard solution—A solution of methyl parahy- buten Hydrate in 20 mL of 0.1 mol/L phosphate buffer solu-
droxybenzoate in diluted acetonitrile (1 in 2) (1 in 1000). tion for antibiotics, pH 8.0. To 4 mL of this solution add 0.1
mol/L phosphate buffer solution for antibiotics, pH 8.0 to
Pipet 5 mL of the sample solution, add 0.1 mol/L phosphate
buffer solution for antibiotics, pH 8.0 to make exactly 100
Ceftibuten Hydrate the test with exactly 5 mL each of the sample solution and
セフチブテン水和物
area of the peak other than ceftibuten obtained from the
ceftibuten obtained from the standard solution, and the total
area of the peaks other than ceftibuten from the sample solu-
tion is not larger than the peak area of ceftibuten from the
C15H14N4O6S2.2H2O: 446.46 standard solution.
(6R,7R)-7-[(2Z )-2-(2-Aminothiazol-4-yl)-4-carboxybut-2- Operating conditions—
enoylamino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- Detector, column, column temperature, mobile phase, and
carboxylic acid dihydrate flow rate: Proceed as directed in the operating conditions in
[118081-34-8] the Assay.
Ceftibuten Hydrate contains not less than 900 mg retention time of ceftibuten, beginning after the solvent
(potency) and not more than 1020 mg (potency) per peak.
mg, calculated on the anhydrous basis. The potency of System suitability—
Ceftibuten Hydrate is expressed as mass (potency) of Test for required detectability: Pipet 2 mL of the standard
ceftibuten (C15H14N4O6S2: 410.42). solution, and add 0.1 mol/L phosphate buffer solution for
antibiotics (pH 8.0) to make exactly 20 mL. Confirm that the
Description Ceftibuten Hydrate occurs as a white to pale
peak area of ceftibuten obtained from 5 mL of this solution
yellowish white crystalline powder.
is equivalent to 7 to 13z of that of ceftibuten obtained from
It is freely soluble in N, N-dimethylformamide and in
dimethyl sulfoxide, and practically insoluble in water, in
System performance: Dissolve 5 mg of Ceftibuten Hydrate
in 20 mL of 0.1 mol/L hydrochloric acid TS, and allow to
Identification (1) Determine the absorption spectrum of a stand at 409C for 1 hour. To 4 mL of this solution add 0.1
solution of Ceftibuten Hydrate in 0.1 mol/L phosphate mol/L phosphate buffer solution for antibiotics (pH 8.0) to
buffer solution for antibiotics, pH 8.0 (1 in 50,000) as di- make 25 mL. When the procedure is run with 5 mL of this so-
rected under Ultraviolet-visible Spectrophotometry <2.24>, lution under the above operating conditions, trans-isomer of
and compare the spectrum with the Reference Spectrum: ceftibuten and ceftibuten are eluted in this order with the
both spectra exhibit similar intensities of absorption at the resolution between these peaks being not less than 2.0.
same wavelengths. System repeatability: When the test is repeated 5 times
(2) Determine the infrared absorption spectrum of with 5 mL of the standard solution under the above operating
Ceftibuten Hydrate as directed in the paste method under conditions, the relative standard deviation of the peak area
the Infrared Spectrophotometry <2.25>, and compare the of ceftibuten is not more than 2.0z.
658 Ceftizoxime Sodium / Official Monographs JP XVII
(ii) Keep the sample solution at not exceeding 59C, and of ceftibuten to that of the internal standard.
use within 24 hours after preparation. To 5 mg of Ceftibuten
Amount [mg (potency)] of ceftibuten (C15H14N4O6S2)
Hydrate add 20 mL of the mobile phase, agitate with the aid
＝ MS × QT/QS × 1000
of ultrasonic waves, if necessary, then shake to dissolve,
filter through a membrane filter with a pore size not MS: Amount [mg (potency)] of Ceftibuten Hydrochloride
exceeding 0.45 mm, and use the filtrate as the sample solu- RS taken
tion. Perform the test with 10 mL of the sample solution as
the following conditions. Determine each peak area by the
automatic integration method, and calculate their amounts
by the area percentage method: the total amount of the
length: 263 nm).
peaks that are eluted faster than ceftibuten is not more than
5.0z. For the areas of these peaks, multiply the relative
response factor, 1.63, respectively.
259C.
length: 263 nm).
Mobile phase: A mixture of 0.005 mol/L n-decyl trimethy-
lammonium bromide TS and acetonitrile (4:1).
ter and 60 cm in length, packed with glycol etherifized silica
Flow rate: Adjust so that the retention time of ceftibuten
259 C.
System performance: Dissolve 5 mg of Ceftibuten Hydrate
Mobile phase: Dissolve 1.05 g of disodium hydrogen phos-
in 20 mL of 0.1 mol/L hydrochloric acid TS, and allow to
phate dodecahydrate and 0.58 g of potassium dihydrogen
stand at 409C for 1 hour. To 4 mL of this solution add 0.1
phosphate in water to make 1000 mL.
mol/L phosphate buffer solution for antibiotics (pH 8.0) to
Flow rate: Adjust so that the retention time of ceftibuten
make 25 mL. When the procedure is run with 5 mL of this so-
lution under the above operating conditions, trans-isomer of
ceftibuten and ceftibuten are eluted in this order with the
retention time of ceftibuten.
resolution between these peaks being not less than 1.5.
lution add the mobile phase to make 20 mL, and use this so-
lution as the solution for system suitability test. Pipet 2 mL
peak area of ceftibuten to that of the internal standard is not
of the solution for system suitability test, and add the mobile
more than 1.0z.
phase to make exactly 20 mL. Confirm that the peak area of
ceftibuten obtained from 10 mL of this solution is equivalent Containers and storage Containers—Tight containers.
to 7 to 13z of that of ceftibuten obtained from 10 mL of the Storage—Light-resistant, and not exceeding 59C.
mL of the solution for system suitability test under the above Ceftizoxime Sodium
the symmetry factor of the peak of ceftibuten are not less セフチゾキシムナトリウム
than 10,000 and 0.8 – 1.2, respectively.
tion of the peak area of ceftibuten is not more than 1.7z.
(0.2 g, volumetric titration, direct titration. Use a mixture of C13H12N5NaO5S2: 405.38
pyridine for water determination and ethylene glycol for Monosodium (6R,7R)-7-[(Z )-2-(2-aminothiazol-4-yl)-2-
water determination (5:1) instead of methanol for water de- (methoxyimino)acetylamino]-8-oxo-5-thia-1-
termination). azabicyclo[4.2.0]oct-2-ene-2-carboxylate
[68401-82-1]
Assay Keep the sample solution and the standard solution Ceftizoxime Sodium contains not less than 925 mg
at not exceeding 59C and use within 2 hours after prepara- (potency) and not more than 965 mg (potency) per mg,
tion. Weigh accurately an amount of Ceftibuten Hydrate calculated on the anhydrous basis. The potency of
and Ceftibuten Hydrochloride RS, equivalent to about 10 Ceftizoxime Sodium is expressed as mass (potency) of
mg (potency), dissolve each in about 36 mL of 0.1 mol/L ceftizoxime (C13H13N5O5S2: 383.40).
phosphate buffer solution for antibiotics (pH 8.0), add ex-
Description Ceftizoxime Sodium occurs as a white to light
actly 4 mL each of the internal standard solution, shake, and
yellow, crystals or crystalline powder.
use these solutions as the sample solution and the standard
It is very soluble in water, sparingly soluble in methanol,
solution, respectively. Perform the test with 5 mL each of the
and practically insoluble in ethanol (95).
uid Chromatography <2.01> according to the following con- Identification (1) Determine the absorption spectrum of a
ditions, and calculate the ratios, QT and QS, of the peak area solution of Ceftizoxime Sodium (1 in 63,000) as directed
JP XVII Official Monographs / Ceftizoxime Sodium 659
under Ultraviolet-visible Spectrophotometry <2.24>, and make exactly 10 mL, and confirm that the peak area of
compare the spectrum with the Reference Spectrum: both ceftizoxime obtained from 5 mL of this solution is equivalent
spectra exhibit similar intensities of absorption at the same to 7 to 13z of that of ceftizoxime obtained from 5 mL of the
wavelengths. solution for test for required detectability.
(2) Determine the infrared absorption spectrum of System performance: Dissolve about 10 mg of
Ceftizoxime Sodium as directed in the potassium bromide Ceftizoxime RS in 100 mL of 0.1 mol/L phosphate buffer
disk method under Infrared Spectrophotometry <2.25>, and solution (pH 7.0) and use this solution as the solution for
compare the spectrum with the Reference Spectrum: both system suitability test. When the procedure is run with 5 mL
spectra exhibit similar intensities of absorption at the same of this solution under the above operating conditions, the
wave numbers. number of theoretical plates and the symmetry factor of the
(3) Determine the 1H spectrum of a solution of peak of ceftizoxime are not less than 4000 and not more than
Ceftizoxime Sodium in heavy water for nuclear magnetic 2.0, respectively.
resonance spectroscopy (1 in 10) as directed under Nuclear System repeatability: When the test is repeated 6 times
Magnetic Resonance Spectroscopy <2.21>, using sodium 3- with 5 mL of the solution for system suitability test under the
trimethylsilylpropionate-d4 for nuclear magnetic resonance above operating conditions, the relative standard deviation
spectroscopy as an internal reference compound: it exhibits a of the peak areas of ceftizoxime is not more than 2.0z.
single signal A at around d 4.0 ppm, a multiple signal B
around d 6.3 ppm, and a single signal C at around d 7.0
ppm. The ratio of integrated intensity of each signal, A:B:C,
is about 3:1:1. Assay Weigh accurately an amount of Ceftizoxime Sodium
(4) Ceftizoxime Sodium responds to the Qualitative and Ceftizoxime RS, equivalent to about 0.1 g (potency),
Tests <1.09> (1) for sodium salt. and dissolve each in 0.1 mol/L phosphate buffer solution
(pH 7.0) to make exactly 20 mL. Pipet 2 mL each of these
D : ＋125 – ＋1459(0.25 g calcu-
solutions, add exactly 10 mL of the internal standard solu-
lated on the anhydrous bases, water, 25 mL, 100 mm).
tion, then add 0.1 mol/L phosphate buffer solution (pH 7.0)
pH <2.54> Dissolve 1.0 g of Ceftizoxime Sodium in 10 mL to make 20 mL, and use these solutions as the sample solu-
of water: the pH of the solution is between 6.0 and 8.0. tion and the standard solution, respectively. Perform the test
of Ceftizoxime Sodium in 10 mL of water: the solution is
clear, and colorless to light yellow.
QS, of the peak area of ceftizoxime to that of the internal
standard.
Ceftizoxime Sodium according to Method 2, and perform
the test. Prepare the control solution with 2.0 mL of Stand- Amount [ mg (potency)] of ceftizoxime (C13H13N5O5S2)
ard Lead Solution (not more than 10 ppm). ＝ MS × QT/QS × 1000
MS: Amount [mg (potency)] of Ceftizoxime RS taken
of Ceftizoxime Sodium according to Method 3, and perform
the test (not more than 1 ppm). Internal standard solution—A solution of 3-hydroxybenzoic
(4) Related substances—Dissolve 0.11 g of Ceftizoxime acid in 0.1 mol/L phosphate buffer solution (pH 7.0) (3 in
Sodium in 100 mL of 0.1 mol/L phosphate buffer solution 500).
(pH 7.0) and use this solution as the sample solution. Per- Operating conditions—
form the test with 5 mL of the sample solution as directed Detector: An ultraviolet absorption photometer (wave-
under Liquid Chromatography <2.01> according to the fol- length: 254 nm).
lowing conditions, and determine the areas of each peak by Column: A stainless steel column 4.6 mm in inside diame-
the automatic integration method: each peak area other than ter and 25 cm in length, packed with octadecylsilanized silica
ceftizoxime is not more than 0.5z of the peak area of gel for liquid chromatography (10 mm in particle diameter).
ceftizoxime, and the total area of peaks other than Column temperature: A constant temperature of about
ceftizoxime is not more than 1.0z of that of ceftizoxime. 359C.
Operating conditions— Mobile phase: Dissolve 2.31 g of disodium hydrogen-
Detector, column, and column temperature: Proceed as phosphate dodecahydrate and 1.42 g of citric acid monohy-
directed in the operating conditions in the Assay. drate in 1000 mL of water, and adjust to pH 3.6 with diluted
Mobile phase: Dissolve 2.31 g of disodium hydrogen- phosphoric acid (1 in 10) or dilute sodium hydroxide TS. To
phosphate dodecahydrate and 1.42 g of citric acid monohy- 450 mL of this solution add 50 mL of acetonitrile.
drate in 1000 mL of water, adjust to pH 3.6 with diluted Flow rate: Adjust so that the retention time of ceftizoxime
phosphoric acid (1 in 10) or dilute sodium hydroxide TS. To is about 4 minutes.
200 mL of this solution add 10 mL of acetonitrile. System suitability—
Flow rate: Adjust so that the retention time of ceftizoxime System performance: When the procedure is run with 5 mL
is about 12 minutes. of the standard solution under the above operating condi-
Time span of measurement: About 5 times as long as the tions, ceftizoxime and the internal standard are eluted in this
retention time of ceftizoxime, beginning after the solvent order with the resolution between these peaks being not less
peak. than 7.0 and the symmetry factor of each peak is not more
System suitability— than 2.
Test for required detectability: Pipet 1 mL of the sample System repeatability: When the test is repeated 6 times
solution, add 0.1 mol/L phosphate buffer solution (pH 7.0) with 5 mL of the standard solution under the above operating
to make exactly 100 mL, and use this solution as the solution conditions, the relative standard deviation of the ratios of
for test for required detectability. Pipet 1 mL of the solu- the peak area of ceftizoxime to that of the internal standard
tion, add 0.1 mol/L phosphate buffer solution (pH 7.0) to is not more than 1.0z.
660 Ceftriaxone Sodium Hydrate / Official Monographs JP XVII
Containers and storage Containers—Tight containers. Ceftriaxone Sodium Hydrate according to Method 2, and
Storage—Light-resistant. perform the test. Prepare the control solution with 2.0 mL of
Ceftriaxone Sodium Hydrate of Ceftriaxone Sodium Hydrate according to Method 3, and
セフトリアキソンナトリウム水和物 (4) Related substances 1—Dissolve 20 mg of Ceftriaxone
Sodium Hydrate in 10 mL of the mobile phase, and use this
lution, add a mixture of water and acetonitrile for liquid
chromatography (11:9) to make exactly 100 mL, and use this
peak area by the automatic integration method: the peak
C18H16N8Na2O7S3.3 1/2 H2O: 661.60
areas of the impurity 1 having the relative retention time of
Disodium (6R,7R)-7-[(Z )-2-(2-aminothiazol-4-yl)-2-
about 0.5 and the impurity 2 having the relative retention
(methoxyimino)acetylamino]-3-(6-hydroxy-2-methyl-5-oxo-
time of about 1.3 to ceftriaxone from the sample solution are
2,5-dihydro-1,2,4-triazin-3-ylsulfanylmethyl)-8-oxo-5-
not larger than the peak area of ceftriaxone from the stand-
ard solution. For the areas of the peaks, the impurity 1 and
hemiheptahydrate
the impurity 2, multiply their relative response factors 0.9
[104376-79-6]
and 1.2, respectively.
Ceftriaxone Sodium Hydrate contains not less than
length: 254 nm).
tency of Ceftriaxone Sodium Hydrate is expressed as
mass (potency) of ceftriaxone (C18H18N8O7S3: 554.58).
Description Ceftriaxone Sodium Hydrate occurs as a white Column temperature: A constant temperature of about
to yellowish white crystalline powder. 259C.
It is freely soluble in water and in dimethylsulfoxide, spar- Mobile phase: Dissolve 5.796 g of anhydrous disodium
ingly soluble in methanol, very slightly soluble in ethanol hydrogen phosphate and 3.522 g of potassium dihydrogen
(99.5), and practically insoluble in acetonitrile. phosphate in water to make exactly 1000 mL, and use this
solution as the solution A. Separately, dissolve 20.256 g of
citric acid monohydrate and 7.840 g of sodium hydroxide in
solution of Ceftriaxone Sodium Hydrate (1 in 100,000) as
solution B. Dissolve 4.00 g of tetra-n-heptylammonium bro-
mide in 450 mL of acetonitrile for liquid chromatography,
the spectrum of a solution of Ceftriaxone Sodium RS pre-
add 55 mL of the solution A, 5 mL of the solution B and 490
pared in the same manner as the sample solution: both spec-
mL of water.
tra exhibit similar intensities of absorption at the same wave-
Flow rate: Adjust so that the retention time of ceftriaxone
lengths.
is about 7 minutes.
Ceftriaxone Sodium Hydrate in deuterated dimethylsul-
retention time of ceftriaxone.
foxide for nuclear magnetic resonance spectroscopy (1 in 10)
as directed under Nuclear Magnetic Resonance Spectroscopy
<2.21>, using tetramethylsilane for nuclear magnetic reso-
lution, add a mixture of water and acetonitrile for liquid
chromatography (11:9) to make 200 mL, and use this solu-
hibits single signals, A, B, C and D, at around d 3.5 ppm, at
tion as the solution for system suitability test. Pipet 1 mL of
around d 3.8 ppm, at around d 6.7 ppm and at around d 7.2
the solution for system suitability test, and add a mixture of
ppm, respectively. The ratio of integrated intensity of each
water and acetonitrile for liquid chromatography (11:9) to
signal, A: B: C: D, is about 3:3:1:2. When the signal at
make exactly 100 mL. Confirm that the peak area of
around d 3.5 ppm overlaps with the signal of water, perform
ceftriaxone obtained from 10 mL of this solution is equiva-
the measurement in the probe kept at about 509 C.
lent to 0.9 to 1.1z of that obtained from 10 mL of the solu-
(3) Ceftriaxone Sodium Hydrate responds to the Quali-
tion for system suitability test.
tative Tests <1.09> (1) for sodium salt.
System performance: Dissolve 10 mg of Ceftriaxone So-
D : －153 – －1709(50 mg calcu- dium Hydrate in a mixture of water and acetonitrile for liq-
lated on the anhydrous basis, water, 2.5 mL, 20 mm). uid chromatography (11:9) to make 5 mL, add 5 mL of a so-
lution of diethyl terephthalate in a mixture of water and
pH <2.54> Dissolve 0.6 g of Ceftriaxone Sodium Hydrate
acetonitrile for liquid chromatography (11:9) (9 in 5000),
in 5 mL of water: the pH of the solution is between 6.0 and
and add a mixture of water and acetonitrile for liquid chro-
8.0.
matography (11:9) to make 200 mL. When the procedure is
Purity (1) Clarity and color of solution—Dissolve 0.6 g run with 10 mL of this solution under the above operating
of Ceftriaxone Sodium Hydrate in 5 mL of water: the solu- conditions, ceftriaxone and diethyl terephthalate are eluted
tion is clear and light yellow. in this order, with the resolution between these peaks being
(2) Heavy metals <1.07>—Proceed with 1.0 g of not less than 6.
JP XVII Official Monographs / Ceftriaxone Sodium Hydrate 661
System repeatability: When the test is repeated 6 times tion of the peak area of ceftriaxone is not more than 1.0z.
tion of the peak area of ceftriaxone is not more than 1.0z.
(5) Related substances 2—Dissolve 10 mg of Ceftriaxone Assay Weigh accurately an amount of Ceftriaxone Sodium
Sodium Hydrate in 10 mL of the mobile phase, and use this Hydrate and Ceftriaxone Sodium RS, equivalent to about
solution as the sample solution. Pipet 1 mL of the sample so- 0.1 g (potency), dissolve each in a mixture of water and
lution, add a mixture of acetonitrile for liquid chromatogra- acetonitrile for liquid chromatography (11:9) to make ex-
phy and water (23:11) to make exactly 100 mL, and use this actly 50 mL. Pipet 5 mL of each solution, add exactly 5 mL
solution as standard solution. Perform the test with exactly of the internal standard solution and a mixture of water and
10 mL each of the sample solution and standard solution as acetonitrile for liquid chromatography (11:9) to make 200
directed under Liquid Chromatography <2.01> according to mL, and use these solutions as the sample solution and the
the following conditions, and determine each peak area by standard solution, respectively. Perform the test with 10 mL
the automatic integration method: the each peak area of the each of the sample solution and standard solution as directed
impurities which appear after the peak of ceftriaxone from under Liquid Chromatography <2.01> according to the fol-
the sample solution is not larger than the peak area of lowing conditions, and calculate the ratios, QT and QS, of
ceftriaxone from the standard solution, and the total peak the peak area of ceftriaxone to that of the internal standard.
area of these impurities is not larger than 2.5 times of the
Amount [ mg (potency)] of ceftriaxone (C18H18N8O7S3)
peak area from the standard solution.
＝ MS × QT/QS × 1000
Detector: An ultraviolet absorption photometer (wave- MS: Amount [mg (potency)] of Ceftriaxone Sodium RS
length: 254 nm). taken
Internal standard solution—A solution of diethyl terephtha-
late in a mixture of water and acetonitrile for liquid chroma-
tography (11:9) (9 in 5000).
259 C.
Mobile phase: Dissolve 5.796 g of anhydrous disodium
length: 254 nm).
phosphate in water to make exactly 1000 mL, and use this
solution as the solution A. Separately, dissolve 20.256 g of
citric acid monohydrate and 7.840 g of sodium hydroxide in
259C.
solution B. Dissolve 4.00 g of tetra-n-heptylammonium bro-
Mobile phase: Dissolve 5.796 g of anhydrous disodium
mide in 450 mL of acetonitrile for liquid chromatography,
and add 55 mL of the solution A, 5 mL of the solution B,
phosphate in water to make exactly 1000 mL, and use this
490 mL of water and 700 mL of acetonitrile for liquid chro-
solution as solution A. Dissolve 20.256 g of citric acid mono-
matography.
hydrate and 7.840 g of sodium hydroxide in water to make
exactly 1000 mL, and use this solution as solution B. Dis-
is about 3 minutes.
solve 4.00 g of tetra-n-heptylammonium bromide in 450 mL
of acetonitrile for liquid chromatography, and add 490 mL
retention time of ceftriaxone.
of water, 55 mL of solution A, and 5 mL of solution B.
Test for required detectability: Measure 5 mL of the sam-
is about 7 minutes.
ple solution, add a mixture of acetonitrile for liquid chroma-
tography and water (23:11) to make 100 mL, and use this so-
lution as the solution for system suitability test. Measure ex-
actly 1 mL of the solution for system suitability test, and add
ditions, ceftriaxone and the internal standard are eluted in
a mixture of acetonitrile for liquid chromatography and
water (23:11) to make exactly 100 mL. Confirm that the
less than 6.
peak area of ceftriaxone obtained from 10 mL of this solu-
tion is equivalent to 0.9 to 1.1z of that obtained from 10 mL
of the solution for system suitability test.
System performance: Dissolve 10 mg of Ceftriaxone So-
of the peak area of ceftriaxone to that of the internal stand-
dium Hydrate in a mixture of acetonitrile for liquid chroma-
tography and water (23:11) to make 5 mL, add 5 mL of a so-
lution of diethyl terephthalate in a mixture of water and Containers and storage Containers—Tight containers.
acetonitrile for liquid chromatography (11:9) (9 in 5000), Storage—Light-resistant.
and add a mixture of acetonitrile for liquid chromatography
and water (23:11) to make 200 mL. When the procedure is
conditions, ceftriaxone and diethyl terephthalate are eluted
not less than 3.
662 Cefuroxime Axetil / Official Monographs JP XVII
Cefuroxime Axetil form the test with exactly 2 mL each of the sample solution
セフロキシムアキセチル raphy <2.01> according to the following conditions, and de-
termine each peak area by the automatic integration method:
the area of the peak other than cefuroxime axetil obtained
from the sample solution is not larger than 1.5 times the
total area of the two peaks of cefuroxime axetil obtained
other than cefuroxime axetil from the sample solution is not
larger than 4 times the total area of the two peaks of
cefuroxime axetil from the standard solution.
C20H22N4O10S: 510.47
(1RS )-1-Acetoxyethyl (6R,7R)-3-carbamoyloxymethyl-7-
[(Z )-2-furan-2-yl-2-(methoxyimino)acetylamino]-8-oxo-5-
the Assay.
[64544-07-6]
retention time of the peak having the larger retention time of
the two peaks of cefuroxime axetil, beginning after the sol-
Cefuroxime Axetil contains not less than 800 mg
vent peak.
calculated on the anhydrous and acetone-free basis.
The potency of Cefuroxime Axetil is expressed as mass
solution, and add 4 mL of methanol and a solution of am-
(potency) of cefuroxime (C16H16N4O8S: 424.39).
monium dihydrogenphosphate (23 in 1000) to make exactly
Description Cefuroxime Axetil occurs as white to yellowish 10 mL. Confirm that the total area of the two peaks of
white, non-crystalline powder. cefuroxime axetil obtained with 2 mL of this solution is
It is freely soluble in dimethylsulfoxide, soluble in metha- equivalent to 7 to 13z of that obtained with 2 mL of the
nol, sparingly soluble in ethanol (95), and very slightly solu- standard solution.
ble in water. System performance: When the procedure is run with 2 mL
tions, the resolution between the two peaks of cefuroxime
solution of Cefuroxime Axetil in methanol (3 in 200,000) as
axetil is not less than 1.5.
the spectrum of a solution of Cefuroxime Axetil RS pre-
conditions, the relative standard deviation of the total area
pared in the same manner as the sample solution: both spec-
of the two peaks of cefuroxime axetil is not more than 2.0z.
tra exhibit similar intensities of absorption at the same wave-
(3) Acetone—Weigh accurately about 1 g of Cefuroxime
lengths.
Axetil, add exactly 0.2 mL of the internal standard solution
and dimethylsulfoxide to make 10 mL, and use this solution
Cefuroxime Axetil as directed in the potassium bromide disk
as the sample solution. Separately, weigh accurately about
0.5 g of acetone, and add dimethylsulfoxide to make exactly
100 mL. Pipet 0.2 mL of this solution, add exactly 0.2 mL of
trum of Cefuroxime Axetil RS: both spectra exhibit similar
the internal standard solution and dimethylsulfoxide to
Cefuroxime Axetil in deuterated dimethylsulfoxide for
standard solution as directed under Gas Chromatography
nuclear magnetic resonance spectroscopy (1 in 20) as directed
under Nuclear Magnetic Resonance Spectroscopy <2.21>,
the ratios, QT and QS, of the peak area of acetone to that of
using tetramethylsilane for nuclear magnetic resonance spec-
the internal standard: the amount of acetone is not more
troscopy as an internal reference compound: it exhibits a
than 1.3z.
double signal or a pair of double signals A at around d 1.5
ppm, a pair of single signals B at around d 2.1 ppm, and a Amount (z) of acetone ＝ MS/MT × QT/QS × 1/5
single signal C at around d 3.9 ppm. The ratio of the inte-
MS: Amount (g) of acetone taken
grated intensity of each signal, A:B:C, is about 1:1:1.
MT: Amount (g) of Cefuroxim Axetil taken
D : ＋41 – ＋479(0.5 g, metha-
Internal standard solution—A solution of 1-propanol in
nol, 50 mL, 100 mm).
dimethylsulfoxide (1 in 200).
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Operating conditions—
Cefuroxime Axetil according to Method 2, and perform the Detector: A hydrogen flame-ionization detector.
test. Prepare the control solution with 2.0 mL of Standard Column: A glass column 3 mm in inside diameter and 2 m
Lead Solution (not more than 10 ppm). in length, packed with siliceous earth for gas chromatogra-
(2) Related substances—Dissolve 25 mg of Cefuroxime phy coated with a mixture of polyethylene glycol 600 for gas
Axetil in 4 mL of methanol, add a solution of ammonium chromatography and polyethylene glycol 1500 for gas chro-
dihydrogenphosphate (23 in 1000) to make 10 mL, and use matography (1:1) in the ratio of 20z (125 – 150 mm in parti-
this solution as the sample solution. Pipet 1 mL of the sam- cle diameter).
ple solution, add 40 mL of methanol and a solution of am- Column temperature: A constant temperature of about
monium dihydrogenphosphate (23 in 1000) to make exactly 909C.
JP XVII Official Monographs / Cellacefate 663
Temperature of injection port: A constant temperature of having the smaller retention time of the two peaks of
about 1159C. cefuroxime axetil is about 8 minutes.
Carrier gas: Nitrogen. System suitability—
Flow rate: Adjust so that the retention time of the internal System performance: When the procedure is run with 10
standard is about 4 minutes. mL of the standard solution under the above operating con-
System suitability— ditions, the internal standard and cefuroxime axetil are
System performance: When the procedure is run with 1 mL eluted in this order with the resolution between the two
of the standard solution under the above operating condi- peaks of cefuroxime axetil being not less than 1.5.
tions, acetone and the internal standard are eluted in this System repeatability: When the test is repeated 6 times
order with the resolution between these peaks being not less with 10 mL of the standard solution under the above
than 5. operating conditions, the relative standard deviation of the
System repeatability: When the test is repeated 6 times ratios of the sum area of the two peaks of cefuroxime
with 1 mL of the standard solution under the above operating axetil to the peak area of the internal standard is not more
conditions, the relative standard deviation of the ratio of the than 1.0z.
peak area of acetone to that of the internal standard is not
more than 5.0z.
Residue on ignition <2.44> Not more than 0.2z (0.5 g). Cellacefate
Isomer ratio Perform the test with 10 mL of the sample so- Cellulose Acetate Phthalate
lution obtained in the Assay as directed under Liquid Chro-
matography <2.01> according to the following conditions, セラセフェート
and determine the area, Aa, of the peak having the smaller
retention time and the area, Ab, of the peak having the big- [9004-38-0]
ger retention time of the two peaks of cefuroxime axetil:
Ab/(Aa ＋ Ab) is between 0.48 and 0.55.
Cellacefate is a reaction product of phthalic anhy-
the Assay.
dride and partially acetylated cellulose.
26.0z of acetyl group (-COCH3: 43.04), and not less
than 30.0z and not more than 36.0z of carboxyben-
Assay Weigh accurately an amount of Cefuroxime Axetil zoyl group (-COC6H4COOH: 149.12), calculated on
and Cefuroxime Axetil RS, equivalent to about 50 mg (po- the anhydrous and free acid-free basis.
tency), and dissolve each in methanol to make exactly 50 
Description Cellacefate occurs as a white powder or
mL. Pipet 10 mL each of these solutions, add exactly 5 mL
grain.
of the internal standard solution, 5 mL of methanol and a
It is freely soluble in acetone, and practically insoluble in
solution of ammonium dihydrogen phosphate (23 in 1000) to
water and in ethanol (99.5).
make 50 mL, and use these solutions as the sample solution
and standard solution. Perform the test with 10 mL each of Identification Determine the infrared absorption spectrum
the sample solution and standard solution as directed under of Cellacefate as directed in the potassium bromide disk
Liquid Chromatography <2.01> according to the following method under Infrared Spectrophotometry <2.25>, and com-
conditions, and calculate the ratios, QT and QS, of the sum pare the spectrum with the Reference Spectrum or the spec-
area of the two peaks of cefuroxime axetil to the peak area trum of Cellacefate RS: both spectra exhibit similar intensi-
of the internal standard. ties of absorption at the same wave numbers.
Amount [ mg (potency)] of cefuroxime (C16H16N4O8S) Viscosity <2.53> Weigh accurately a quantity of Cellace-
＝ MS × QT/QS × 1000 fate, equivalent to 15 g calculated on the anhydrous basis,
dissolve in 85 g of a mixture of acetone and water (249: 1 in
MS: Amount [mg (potency)] of Cefuroxime Axetil RS
mass), and use this solution as the sample solution. Perform
taken
the test with the sample solution at 25 ± 0.29C as directed in
Internal standard solution—A solution of acetanilide in Method 1 to obtain the kinematic viscosity n. Separately, de-
methanol (27 in 5000). termine the density, r, of the sample solution as directed
Operating conditions— under Determination of Specific Gravity and Density <2.56>,
Detector: An ultraviolet absorption photometer (wave- and calculate the viscosity of the sample solution, h, as h ＝
length: 278 nm). rn: not less than 45 mPa･s and not more than 90 mPa･s.
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
ter and 20 cm in length, packed with trimethylsilanized silica
Cellacefate according to Method 2, and perform the test.
Solution (not more than 10 ppm).
259 C.
(2) Free acids—Weigh accurately about 3 g of Cellace-
Mobile phase: A mixture of a solution of ammonium di-
fate, put in a glass-stoppered conical flask, add 100 mL of
hydrogen phosphate (23 in 1000) and methanol (5:3).
diluted methanol (1 in 2), stopper tightly, and filter after
Flow rate: Adjust so that the retention time of the peak
shaking for 2 hours. Wash both the flask and residue with
664 Microcrystalline Cellulose / Official Monographs JP XVII
two 10-mL portions each of diluted methanol (1 in 2), com-
bine the washes to the filtrate, and titrate <2.50> with 0.1 Microcrystalline Cellulose
mol/L sodium hydroxide VS (indicator: 2-3 drops of phenol-
phthalein TS). Perform the blank determination with 120 結晶セルロース
mL of diluted methanol (1 in 2), and make any necessary
correction. [9004-34-6, cellulose]
Amount (z) of free acids ＝ 0.8306A/M
A: Amount (mL) of 0.1 mol/L sodium hydroxide VS con- macopoeia and the U.S. Pharmacopeia. The parts of the text
sumed that are not harmonized are marked with symbols ( ).
M: Amount (g) of Cellacefate taken, calculated on the an-
hydrous basis Microcrystalline Cellulose is purified, partially
depolymerized a-cellulose, obtained as a pulp from fi-
The amount of free acids is not more than 3.0z, calcu-
brous plant material, with mineral acids.
lated as phthalic acid (C8H6O4: 166.13). 
The label indicates the mean degree of polymeriza-
Water <2.48> Not more than 5.0z (0.5 g, volumetric titration, loss on drying, and bulk density values with a
tion, direct titration, using a mixture of ethanol (99.5) and range.
dichloromethane (3:2) instead of methanol for water deter- Description Microcrystalline Cellulose occurs as a white
mination).
crystalline powder having fluidity.
Residue on ignition <2.44> Not more than 0.1z (1 g). It is practically insoluble in water, in ethanol (95) and in
diethyl ether.
Assay (1) Carboxybenzoyl group—Weigh accurately about
It swells with sodium hydroxide TS on heating.
1 g of Cellacefate, dissolve in 50 mL of a mixture of ethanol
(95) and acetone (3:2), and titrate <2.50> with 0.1 mol/L so- Identification (1) Dissolve 20 g of zinc chloride and 6.5 g
dium hydroxide VS (indicator: 2 – 3 drops of phenolphthal- of potassium iodide in 10.5 mL of water, add 0.5 g of iodine,
ein TS). Perform a blank determination, and make any nec- and shake for 15 minutes. Place about 10 mg of
essary correction. Microcrystalline Cellulose on a watch glass, and disperse in
2 mL of this solution: the substance develops a blue-violet
Content (z) of carboxybenzoyl group (C8H5O3)
color.
1.491 × A (2) Sieve 20 g of Microcrystalline Cellulose for 5
－(1.795 × B)
M minutes on an air-jet sieve equipped with a screen (No.391,
＝ ×100
100 － B 200 mm in inside diameter) having 38-mm openings. If more
than 5z is retained on the screen, mix 30 g of Microcrystal-
A: Amount (mL) of 0.1 mol/L sodium hydroxide VS con-
line Cellulose with 270 mL of water; otherwise, mix 45 g
sumed
with 255 mL of water. Perform the mixing for 5 minutes in a
B: Amount (z) of free acids obtained in the Purity (2)
high-speed (18,000 revolutions per minute or more) power
M: Amount (g) of Cellacefate taken, calculated on the an-
blender. Transfer 100 mL of the dispersion to a 100-mL
hydrous basis
graduated cylinder, and allow to stand for 3 hours: a white,
(2) Acetyl group—Weigh accurately about 0.1 g of Cel- opaque, bubble-free dispersion, which does not form a su-
lacefate, put in a glass-stoppered conical flask, add exactly pernatant liquid at the surface, is obtained.
25 mL of 0.1 mol/L sodium hydroxide VS, and boil for 30 (3) Transfer 1.3 g of Microcrystalline Cellulose, accu-
minutes under a reflux condenser. After cooling, add 2 – 3 rately weighed, to a 125-mL conical flask, and add exactly 25
drops of phenolphthalein TS, and titrate <2.50> the excess of mL each of water and 1 mol/L cupriethylenediamine TS.
sodium hydroxide with 0.1 mol/L hydrochloric acid VS. Immediately purge the solution with nitrogen, insert the
Perform a blank determination. stopper, and shake on a suitable mechanical shaker to dis-
solve. Perform the test with a suitable amount of this solu-
Content (z) of free acids and bound acetyl group (C2H3O)
tion, taken exactly, according to Method 1 under Viscosity
＝ 0.4305A/M
Determination <2.53> using a capillary viscometer having the
A: Amount (mL) of 0.1 mol/L sodium hydroxide VS con- viscosity constant (K) of approximately 0.03, at 25 ± 0.19 C,
sumed, corrected by the blank determination and determine the kinematic viscosity, n. Separately, per-
M: Amount (g) of Cellacefate taken, calculated on the an- form the test with a mixture of exactly 25 mL each of water
hydrous basis and 1 mol/L cupriethylenediamine TS in the same manner as
above, using a capillary viscometer having K of approxi-
Content (z) of acetyl group (C2H3O)
mately 0.01, and determine the kinematic viscosity, no.
＝100 × (P － 0.5182B)/(100 － B) － 0.5772C
Calculate the relative viscosity, hrel, of Microcrystalline
B: Amount (z) of free acids obtained in the Purity (2) Cellulose by the formula:
C: Content (z) of carboxybenzoyl group
hrel ＝ n/no
P: Content (z) of free acids and bound acetyl group
(C2H3O) Obtain the product, [h]C, of intrinsic viscosity [h](mL/g)
 and concentration C (g/100 mL) from the value hrel of the
Containers and storage Containers—Tight containers.
Table. When calculate the degree of polymerization, P, by
the following formula, P is not more than 350 and within
the labeled range.
P ＝ (95)[h]C/MT
MT: Amount (g) of the sample taken, calculated on the
dried basis
JP XVII Official Monographs / Microcrystalline Cellulose 665
pH <2.54> Shake 5.0 g of Microcrystalline Cellulose with

40 mL of water for 20 minutes, and centrifuge: the pH of the
supernatant liquid is between 5.0 and 7.5.
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
Microcrystalline Cellulose according to Method 2, and per-
Standard Lead Solution (not more than 10 ppm).
(2) Water-soluble substances—Shake 5.0 g of
Microcrystalline Cellulose with 80 mL of water for 10
minutes, filter with the aid of vacuum through a filter paper
for quantitative analysis (5C) into a vacuum flask.
Evaporate the clear filtrate in a tared evaporating dish to
dryness without charring, dry at 1059 C for 1 hour, cool in a
desiccator, and weigh: the difference between the mass of
the residue and the mass obtained from a blank determina-
tion does not exceed 12.5 mg.
as it passes. At the bottom of the baffle box is a funnel that
(3) Diethyl ether-soluble substances—Place 10.0 g of
collect the powder, and allows it to pour into a sample
Microcrystalline Cellulose in a column having an internal di-
receiving cup mounted directly below it.
ameter of about 20 mm, and pass 50 mL of peroxide-free
(ii) Procedure—Weigh accurately the mass of a brass or
diethyl ether through the column. Evaporate the eluate to
stainless steel cup, which has a capacity of 25.0 ± 0.05 mL
dryness in a previously dried and tared evaporation dish.
and an inside diameter of 30.0 ± 2.0 mm, and put the cup
Dry the residue at 1059C for 30 minutes, allow to cool in a
directly below the funnel of the volumeter. Slowly pour
desiccator, and weigh: the difference between the mass of
Microcrystalline Cellulose 5.1 cm height from the upper part
the residue and the mass obtained from a blank determina-
of the powder funnel through the sieve, at a rate suitable to
tion does not exceed 5.0 mg.
prevent clogging, until the cup overflows. If the clogging oc-
Conductivity <2.51> Perform the test as directed in the curs, take out the sieve. Level the excess powder with the aid
Conductivity Measurement with the supernatant liquid ob- of a slide glass, weigh the filled cup, and weigh accurately
tained in the pH as the sample solution, and determine the the content of the cup, and then calculate the bulk density by
conductivity at 25 ± 0.19 C. Determine in the same way the following expression: the bulk density is within the
the conductivity of water used for the preparation of the labeled specification.
sample solution: the difference between these conductivities
Bulk density (g/cm3 ) ＝ A/25
is not more than 75 mS･cm－1.
A: Measured mass (g) of the content of the cup
Loss on drying <2.41> Not more than 7.0z and within a
range as specified on the label (1 g, 1059C. 3 hours). Microbial limit <4.05> The acceptance criteria of TAMC
 and TYMC are 103 CFU/g and 102 CFU/g, respectively. Es-
Residue on ignition <2.44> Not more than 0.1z (2 g).
cherichia coli, Salmonella, Pseudomonas aeruginosa and
Bulk density (i) Apparatus—Use a volumeter shown in Staphylococcus aureus are not observed.
the figure. Put a No.8.6 sieve (2000 mm) on the top of the Containers and storage Containers—Tight containers.
volumeter. A funnel is mounted over a baffle box, having
four glass baffle plates inside which the sample powder slides
Table for Conversion of Relative Viscosity ( hrel) into the Product of Limiting Viscosity and Concentration ([h]C)
[ h]C
hrel 0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
1.1 0.098 0.106 0.115 0.125 0.134 0.143 0.152 0.161 0.170 0.180
1.2 0.189 0.198 0.207 0.216 0.225 0.233 0.242 0.250 0.259 0.268
1.3 0.276 0.285 0.293 0.302 0.310 0.318 0.326 0.334 0.342 0.350
1.4 0.358 0.367 0.375 0.383 0.391 0.399 0.407 0.414 0.422 0.430
1.5 0.437 0.445 0.453 0.460 0.468 0.476 0.484 0.491 0.499 0.507
1.6 0.515 0.522 0.529 0.536 0.544 0.551 0.558 0.566 0.573 0.580
1.7 0.587 0.595 0.602 0.608 0.615 0.622 0.629 0.636 0.642 0.649
1.8 0.656 0.663 0.670 0.677 0.683 0.690 0.697 0.704 0.710 0.717
1.9 0.723 0.730 0.736 0.743 0.749 0.756 0.762 0.769 0.775 0.782
2.0 0.788 0.795 0.802 0.809 0.815 0.821 0.827 0.833 0.840 0.846
2.1 0.852 0.858 0.864 0.870 0.876 0.882 0.888 0.894 0.900 0.906
2.2 0.912 0.918 0.924 0.929 0.935 0.941 0.948 0.953 0.959 0.965
2.3 0.971 0.976 0.983 0.988 0.994 1.000 1.006 1.011 1.017 1.022
2.4 1.028 1.033 1.039 1.044 1.050 1.056 1.061 1.067 1.072 1.078
2.5 1.083 1.089 1.094 1.100 1.105 1.111 1.116 1.121 1.126 1.131
2.6 1.137 1.142 1.147 1.153 1.158 1.163 1.169 1.174 1.179 1.184
2.7 1.190 1.195 1.200 1.205 1.210 1.215 1.220 1.225 1.230 1.235
2.8 1.240 1.245 1.250 1.255 1.260 1.265 1.270 1.275 1.280 1.285
2.9 1.290 1.295 1.300 1.305 1.310 1.314 1.319 1.324 1.329 1.333
666 Microcrystalline Cellulose / Official Monographs JP XVII
[ h]C
hrel 0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
3.0 1.338 1.343 1.348 1.352 1.357 1.362 1.367 1.371 1.376 1.381
3.1 1.386 1.390 1.395 1.400 1.405 1.409 1.414 1.418 1.423 1.427
3.2 1.432 1.436 1.441 1.446 1.450 1.455 1.459 1.464 1.468 1.473
3.3 1.477 1.482 1.486 1.491 1.496 1.500 1.504 1.508 1.513 1.517
3.4 1.521 1.525 1.529 1.533 1.537 1.542 1.546 1.550 1.554 1.558
3.5 1.562 1.566 1.570 1.575 1.579 1.583 1.587 1.591 1.595 1.600
3.6 1.604 1.608 1.612 1.617 1.621 1.625 1.629 1.633 1.637 1.642
3.7 1.646 1.650 1.654 1.658 1.662 1.666 1.671 1.675 1.679 1.683
3.8 1.687 1.691 1.695 1.700 1.704 1.708 1.712 1.715 1.719 1.723
3.9 1.727 1.731 1.735 1.739 1.742 1.746 1.750 1.754 1.758 1.762
4.0 1.765 1.769 1.773 1.777 1.781 1.785 1.789 1.792 1.796 1.800
4.1 1.804 1.808 1.811 1.815 1.819 1.822 1.826 1.830 1.833 1.837
4.2 1.841 1.845 1.848 1.852 1.856 1.859 1.863 1.867 1.870 1.874
4.3 1.878 1.882 1.885 1.889 1.893 1.896 1.900 1.904 1.907 1.911
4.4 1.914 1.918 1.921 1.925 1.929 1.932 1.936 1.939 1.943 1.946
4.5 1.950 1.954 1.957 1.961 1.964 1.968 1.971 1.975 1.979 1.982
4.6 1.986 1.989 1.993 1.996 2.000 2.003 2.007 2.010 2.013 2.017
4.7 2.020 2.023 2.027 2.030 2.033 2.037 2.040 2.043 2.047 2.050
4.8 2.053 2.057 2.060 2.063 2.067 2.070 2.073 2.077 2.080 2.083
4.9 2.087 2.090 2.093 2.097 2.100 2.103 2.107 2.110 2.113 2.116
5.0 2.119 2.122 2.125 2.129 2.132 2.135 2.139 2.142 2.145 2.148
5.1 2.151 2.154 2.158 2.160 2.164 2.167 2.170 2.173 2.176 2.180
5.2 2.183 2.186 2.190 2.192 2.195 2.197 2.200 2.203 2.206 2.209
5.3 2.212 2.215 2.218 2.221 2.224 2.227 2.230 2.233 2.236 2.240
5.4 2.243 2.246 2.249 2.252 2.255 2.258 2.261 2.264 2.267 2.270
5.5 2.273 2.276 2.279 2.282 2.285 2.288 2.291 2.294 2.297 2.300
5.6 2.303 2.306 2.309 2.312 2.315 2.318 2.320 2.324 2.326 2.329
5.7 2.332 2.335 2.338 2.341 2.344 2.347 2.350 2.353 2.355 2.358
5.8 2.361 2.364 2.367 2.370 2.373 2.376 2.379 2.382 2.384 2.387
5.9 2.390 2.393 2.396 2.400 2.403 2.405 2.408 2.411 2.414 2.417
6.0 2.419 2.422 2.425 2.428 2.431 2.433 2.436 2.439 2.442 2.444
6.1 2.447 2.450 2.453 2.456 2.458 2.461 2.464 2.467 2.470 2.472
6.2 2.475 2.478 2.481 2.483 2.486 2.489 2.492 2.494 2.497 2.500
6.3 2.503 2.505 2.508 2.511 2.513 2.516 2.518 2.521 2.524 2.526
6.4 2.529 2.532 2.534 2.537 2.540 2.542 2.545 2.547 2.550 2.553
6.5 2.555 2.558 2.561 2.563 2.566 2.568 2.571 2.574 2.576 2.579
6.6 2.581 2.584 2.587 2.590 2.592 2.595 2.597 2.600 2.603 2.605
6.7 2.608 2.610 2.613 2.615 2.618 2.620 2.623 2.625 2.627 2.630
6.8 2.633 2.635 2.637 2.640 2.643 2.645 2.648 2.650 2.653 2.655
6.9 2.658 2.660 2.663 2.665 2.668 2.670 2.673 2.675 2.678 2.680
7.0 2.683 2.685 2.687 2.690 2.693 2.695 2.698 2.700 2.702 2.705
7.1 2.707 2.710 2.712 2.714 2.717 2.719 2.721 2.724 2.726 2.729
7.2 2.731 2.733 2.736 2.738 2.740 2.743 2.745 2.748 2.750 2.752
7.3 2.755 2.757 2.760 2.762 2.764 2.767 2.769 2.771 2.774 2.776
7.4 2.779 2.781 2.783 2.786 2.788 2.790 2.793 2.795 2.798 2.800
7.5 2.802 2.805 2.807 2.809 2.812 2.814 2.816 2.819 2.821 2.823
7.6 2.826 2.828 2.830 2.833 2.835 2.837 2.840 2.842 2.844 2.847
7.7 2.849 2.851 2.854 2.856 2.858 2.860 2.863 2.865 2.868 2.870
7.8 2.873 2.875 2.877 2.879 2.881 2.884 2.887 2.889 2.891 2.893
7.9 2.895 2.898 2.900 2.902 2.905 2.907 2.909 2.911 2.913 2.915
8.0 2.918 2.920 2.922 2.924 2.926 2.928 2.931 2.933 2.935 2.937
8.1 2.939 2.942 2.944 2.946 2.948 2.950 2.952 2.955 2.957 2.959
8.2 2.961 2.963 2.966 2.968 2.970 2.972 2.974 2.976 2.979 2.981
8.3 2.983 2.985 2.987 2.990 2.992 2.994 2.996 2.998 3.000 3.002
8.4 3.004 3.006 3.008 3.010 3.012 3.015 3.017 3.019 3.021 3.023
8.5 3.025 3.027 3.029 3.031 3.033 3.035 3.037 3.040 3.042 3.044
8.6 3.046 3.048 3.050 3.052 3.054 3.056 3.058 3.060 3.062 3.064
8.7 3.067 3.069 3.071 3.073 3.075 3.077 3.079 3.081 3.083 3.085
8.8 3.087 3.089 3.092 3.094 3.096 3.098 3.100 3.102 3.104 3.106
8.9 3.108 3.110 3.112 3.114 3.116 3.118 3.120 3.122 3.124 3.126
9.0 3.128 3.130 3.132 3.134 3.136 3.138 3.140 3.142 3.144 3.146
9.1 3.148 3.150 3.152 3.154 3.156 3.158 3.160 3.162 3.164 3.166
9.2 3.168 3.170 3.172 3.174 3.176 3.178 3.180 3.182 3.184 3.186
9.3 3.188 3.190 3.192 3.194 3.196 3.198 3.200 3.202 3.204 3.206
9.4 3.208 3.210 3.212 3.214 3.215 3.217 3.219 3.221 3.223 3.225
9.5 3.227 3.229 3.231 3.233 3.235 3.237 3.239 3.241 3.242 3.244
JP XVII Official Monographs / Powdered Cellulose 667
[ h]C
hrel 0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
9.6 3.246 3.248 3.250 3.252 3.254 3.256 3.258 3.260 3.262 3.264
9.7 3.266 3.268 3.269 3.271 3.273 3.275 3.277 3.279 3.281 3.283
9.8 3.285 3.287 3.289 3.291 3.293 3.295 3.297 3.298 3.300 3.302
9.9 3.304 3.305 3.307 3.309 3.311 3.313 3.316 3.318 3.320 3.321
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
10 3.32 3.34 3.36 3.37 3.39 3.41 3.43 3.45 3.46 3.48
11 3.50 3.52 3.53 3.55 3.56 3.58 3.60 3.61 3.63 3.64
12 3.66 3.68 3.69 3.71 3.72 3.74 3.76 3.77 3.79 3.80
13 3.80 3.83 3.85 3.86 3.88 3.89 3.90 3.92 3.93 3.95
14 3.96 3.97 3.99 4.00 4.02 4.03 4.04 4.06 4.07 4.09
15 4.10 4.11 4.13 4.14 4.15 4.17 4.18 4.19 4.20 4.22
16 4.23 4.24 4.25 4.27 4.28 4.29 4.30 4.31 4.33 4.34
17 4.35 4.36 4.37 4.38 4.39 4.41 4.42 4.43 4.44 4.45
18 4.46 4.47 4.48 4.49 4.50 4.52 4.53 4.54 4.55 4.56
19 4.57 4.58 4.59 4.60 4.61 4.62 4.63 4.64 4.65 4.66
Lead Solution (not more than 10 ppm).

Powdered Cellulose (2) Water-soluble substances—Shake 6.0 g of Powdered
Cellulose with 90 mL of recently boiled and cooled water,
粉末セルロース and allow to stand for 10 minutes with occasional shaking.
Filter, with the aid of vacuum through a filter paper, discard
[9004-34-6, Cellulose] the first 10 mL of the filtrate, and pass the subsequent fil-
trate through the same filter, if necessary, to obtain a clear
This monograph is harmonized with the European Phar- filtrate. Evaporate a 15.0-mL portion of the filtrate in a
macopoeia and the U.S. Pharmacopeia. The parts of the text tared evaporating dish to dryness without charring, dry at
that are not harmonized are marked with symbols ( ). 1059C for 1 hour, and weigh after allowing to cool in a
desiccator: the difference between the mass of the residue
Powdered Cellulose is a purified, mechanically dis- and the mass obtained from a blank determination does not
integrated alpha cellulose obtained as a pulp, after exceed 15.0 mg (1.5z).
partial hydrolysis as occasion demands, from fibrous (3) Diethyl ether-soluble substances—Place 10.0 g of
plant materials. Powdered Cellulose in a column having an internal diameter
The label indicates the mean degree of polymeriza- of about 20 mm, and pass 50 mL of peroxide-free diethyl
tion value with a range. ether through the column. Evaporate the eluate to dryness in
Description a previously dried and tared evaporation dish. Dry the
Powdered Cellulose occurs as a white pow-
residue at 1059C for 30 minutes, and weigh after allowing to
der.
cool in a desiccator: the difference between the mass of the
residue and the mass obtained from a blank determination
diethyl ether.
does not exceed 15.0 mg (0.15z).
Identification (1) Dissolve 20 g of zinc chloride and 6.5 g
of potassium iodide in 10.5 mL of water, add 0.5 g of iodine,
3 hours).
and shake for 15 minutes. Place about 10 mg of Powdered
Cellulose on a watch glass, and disperse in 2 mL of this solu- Residue on ignition <2.44> Not more than 0.3z (1 g calcu-
tion: the substance develops a blue-violet color. lated on the dried basis).
(2) Mix 30 g of Powdered Cellulose with 270 mL of Microbial limit <4.05> The acceptance criteria of TAMC
water in a high-speed (18,000 revolutions per minute or
and TYMC are 103 CFU/g and 102 CFU/g, respectively. Es-
more) blender for 5 minutes, transfer 100 mL of the disper-
cherichia coli, Salmonella, Pseudomonas aeruginosa and
sion to a 100-mL graduated cylinder, and allow to stand for
Staphylococcus aureus are not observed.
1 hour: a supernatant liquid appears above the layer of the
cellulose. Containers and storage Containers—Tight containers.
(3) Transfer 0.25 g of Powdered Cellulose, accurately
weighed, to a 125-mL conical flask, add exactly 25 mL each
of water and 1 mol/L cupriethylenediamine TS, and proceed
as directed in the Identification (3) under Microcrystalline
Cellulose. The mean degree of polymerization, P, is not less
than 440 and is within the labeled specification.
pH <2.54> Mix 10 g of Powdered Cellulose with 90 mL of
water, and allow to stand for 1 hour with occasional stirring:
the pH of the supernatant liquid is between 5.0 and 7.5.
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
Powdered Cellulose according to Method 2, and perform the
668 Celmoleukin (Genetical Recombination) / Official Monographs JP XVII
379C for 2 hours, and then run this solution under the above
Celmoleukin (Genetical operating conditions. Celmoleukin and its reduced form are
Recombination) being not less than 1.5.
(2) Accurately measure an appropriate amount of Cel-
セルモロイキン（遺伝子組換え）
moleukin (Genetical Recombination), dilute by adding cul-
ture medium for celmoleukin, and prepare a sample solution
containing 800 units per mL. Add 25 mL of the sample solu-
tion to 2 wells (A and B) of a flat-bottomed microtest plate
for tissue culture, and then add 25 mL of reference anti-inter-
leukin-2 antiserum solution diluted with culture medium for
celmoleukin to well A and 25 mL of culture medium for cel-
C693H1118N178O203S7: 15415.82
moleukin to well B. Add 50 mL of culture medium for cel-
[94218-72-1]
moleukin to another well (well C). After shaking the
microtest plate, warm in air containing 5z carbon dioxide at
Celmoleukin (Genetical Recombination) is genetical
379C for 30 minutes to 2 hours. Next, add to each well 50 mL
recombinant human interleukin-2, and is a protein
of culture medium for celmoleukin containing the interleu-
consisting of 133 amino acid residues. It is a solution.
kin-2 dependent mouse natural killer cells NKC3 and culture
It has a T-lymphocyte activating effect.
at 379 C for 16 to 24 hours. Add 3-(4,5-dimethylthiazole-
It contains not less than 0.5 and not more than 1.5
2-yl)-2,5-diphenyl-2H-tetrazolium bromide TS, culture at
mg of protein per mL, and 1 mg of this protein con-
379C for 4 to 6 hours, and add sodium lauryl sulfate TS and
tains potency not less than 8.0 × 106 units.
leave for 24 to 48 hours. When the absorbance at 590 nm of
Description Celmoleukin (Genetical Recombination) oc- the solution in each well is measured, the difference in absor-
curs as a clear and colorless liquid. bance between the solutions from wells A and C is not more
than 3z of the difference in absorbance between the solu-
Identification (1) Add 100 mL of protein digestive enzyme
tions from wells B and C.
TS to 100 mL of Celmoleukin (Genetical Recombination),
shake, leave standing at 379 C for 18 to 24 hours, and then Constituent amino acid When hydrolyze Celmoleukin
add 2 mL of 2-mercaptoethanol. Leave at 379C for a further (Genetical Recombination) according to Method 1 and
30 minutes, and add 5 mL of trifluoroacetic acid solution (1 Method 4 described in ``1. Hydrolysis of Protein and Pep-
in 10). Use this solution as the sample solution. Separately, tide'', and perform the test according to Method 1 described
process with celmoleukin for liquid chromatography by in ``2. Methodologies of Amino Acid Analysis'' under Ami-
using the same method. Use this solution as the standard so- no Acid Analysis of Proteins <2.04>, the molar ratios of the
lution. Perform the test with 50 mL each of the sample solu- respective amino acids are as follows: glutamic acid (or
tion and standard solution as directed under Liquid Chroma- glutamine) is 17 or 18, threonine is 11 to 13, aspartic acid (or
tography <2.01> according to the following conditions, and asparagine) is 11 or 12, lysine is 11, isoleucine is 7 or 8, ser-
compare the chromatograms obtained from the sample solu- ine is 6 to 9, phenylalanine is 6, alanine is 5, proline is 5 or 6,
tion and standard solution: the similar peaks are observed at arginine is 4, methionine is 4, cysteine is 3 or 4, valine is 3 or
the same retention time. 4, tyrosine is 3, histidine is 3, glycine is 2, and tryptophan is
Operating conditions— 1.
Detector: An ultraviolet absorption photometer (wave- Procedure
length: 215 nm). (i) Hydrolysis Based on the results of the Assay (1),
Column: A stainless steel column 4 mm in inside diameter place an amount of Celmoleukin (Genetical Recombination),
and 30 cm in length, packed with octadecylsilanized silica gel equivalent to about 50 mg as the total protein in two hydroly-
for liquid chromatography (particle size: 5 mm). sis tubes, and evaporate to dryness under vacuum. To one of
Column temperature: A constant temperature of about the hydrolysis tubes add 100 mL of a mixture of diluted hy-
259 C. drochloric acid (59 in 125), mercapto acetic acid and phenol
Mobile phase A: A solution of trifluoroacetic acid (1 in (100:10:1), and shake. Place this hydrolysis tube in a vial and
1000). humidify the inside of the vial with 200 mL of the mixture of
Mobile phase B: A solution of trifluoroacetic acid in a diluted hydrochloric acid (59 in 125), mercapto acetic acid
mixture of acetonitrile and water (17:3) (1 in 1000). and phenol (100:10:1). Replace the vial interior with inert
Flowing of mobile phase: Control the gradient by mixing gas or reduce the pressure, and heat at about 1159C for 24
the mobile phases A and B as directed in the following table. hours. After drying under vacuum, dissolve in 0.5 mL of
0.02 mol/L hydrochloric acid TS, and use this solution as
Time after injection Mobile phase A Mobile phase B the sample solution (1). To the other hydrolysis tube, add
of sample (min) (volz) (volz) 100 mL of ice cold performic acid, oxidize for 1.5 hours on
ice, add 50 mL of hydrobromic acid, and dry under vacuum.
0–5 100 0 Add 200 mL of water, repeat the dry under vacuum proce-
5 – 45 100 → 60 0 → 40 dure two more times, place the hydrolysis tube in a vial, and
45 – 75 60 → 0 40 → 100 humidify the inside of the vial with 200 mL of diluted hydro-
75 – 85 0 100 chloric acid (59 in 125). Replace the vial interior with inert
gas or reduce the pressure, and heat at about 1159C for 24
Flow rate: Adjust so that the retention time of celmoleu- hours. After drying under vacuum, dissolve in 0.5 mL of
kin is about 70 minutes. 0.02 mol/L hydrochloric acid TS, and use this solution as
System suitability— the sample solution (2). Separately, weigh exactly 60 mg of
System performance: Add 2 mL of 2-mercaptoethanol to L-aspartic acid, 100 mg of L-glutamic acid, 17 mg of L-ala-
100 mL of celmoleukin for liquid chromatography, leave at nine, 23 mg of L-methionine, 21 mg of L-tyrosine, 24 mg of
JP XVII Official Monographs / Celmoleukin (Genetical Recombination) 669
L-histidine hydrochloride monohydrate, 58 mg of L-threo-

nine, 22 mg of L-proline, 14 mg of L-cystine, 45 mg of L- Time after Mobile Mobile Mobile Mobile
injection of phase A phase B phase C phase D
isoleucine, 37 mg of L-phenylalanine, 32 mg of L-arginine
sample (min) (volz) (volz) (volz) (volz)
hydrochloride, 32 mg of L-serine, 6 mg of glycine, 18 mg of
L-valine, 109 mg of L-leucine, 76 mg of L-lysine hydrochlo- 0 – 35 100 0 0 0
ride, and 8 mg of L-tryptophan, dissolve with 0.1 mol/L hy- 35 – 60 0 100 0 0
drochloric acid TS to make exactly 500 mL, and use this so- 60 – 111 0 0 100 0
lution as the standard solution. Transfer 40 mL each of the 111 – 121 0 0 0 100
standard solution to two hydrolysis tubes, evaporate to dry-
ness under vacuum, and proceed in the same way for each Reaction reagent: Mix 407 g of lithium acetate dihydrate,
respective sample solution to make the standard solutions (1) 245 mL of acetic acid (100) and 801 mL of 1-methoxy-2-
and (2). propanol, add water to make 2000 mL, stir for 10 minutes
(ii) Amino acid analysis Perform the test with exactly while passing a current of nitrogen, and assign as solution A.
250 mL each of the sample solutions (1) and (2) and standard Separately, to 1957 mL of 1-methoxy-2-propanol add 77 g of
solutions (1) and (2) as directed under Liquid Chromatogra- ninhydrin and 0.134 g of sodium borohydride, stir for 30
phy <2.01> according to the following conditions, and from minutes while passing a current of nitrogen, and assign as
the peak areas for each amino acid obtained from the sample solution B. Mix solutions A and B before use.
solutions (1) and (2) and standard solutions (1) and (2) calcu- Flow rate of mobile phase: Adjust so that the retention
late the molar number of the amino acids contained in 1 mL times of serine and leucine are about 30 minutes and about
of the sample solutions (1) and (2). Furthermore, calculate 73 minutes, respectively (about 0.21 mL per minute).
the number of amino acids assuming there are 22 leucine Flow rate of reaction reagent: About 0.25 mL per minute.
residues in one mole of celmoleukin. System suitability—
Operating conditions— System performance: To 2 mL of the standard solution
Detector: A visible absorption photometer [wavelength: add 0.02 mol/L hydrochloric acid TS to make 25 mL. When
440 nm (proline) and 570 nm (amino acids other than pro- the procedure is run with 250 mL of this solution under the
line)]. above operating conditions, the resolution between the peaks
Column: A stainless steel column 4 mm in inside diameter of threonine and serine is not less than 1.2.
and 25 cm in length, packed with strongly acidic ionex- System repeatability: To 2 mL of the standard solution
change resin for liquid chromatography (Na type) (sulfonic add 0.02 mol/L hydrochloric acid TS to make 25 mL. When
acid group bound divinylbenzenepolystyrene) (5 mm in parti- the test is repeated 3 times with 250 mL of this solution under
cle diameter). the above operating conditions, the relative standard devia-
Column temperature: Maintaining a constant temperature tion of the peak area of aspartic acid, serine, arginine and
of about 489C for 28 minutes after sample injection, then a proline is not more than 2.4z.
constant temperature of about 629 C until 121 minutes after
Molecular mass Based on the results of the Assay (1), add
the injection.
buffer for celmoleukin and dilute to prepare a sample solu-
Reaction temperature: A constant temperature of about
tion so that there is about 0.5 mg of protein per mL. To ver-
1359C.
tical uncontinuous buffer SDS-polyacrylamide gel prepared
Color developing time: About 1 minute.
from resolving gel for celmoleukin and stacking gel for cel-
Mobile phases A, B, C and D: Prepare according to the
moleukin add 20 mL of the sample solution or 20 mL of mar-
following table.
ker protein for celmoleukin molecular mass determination to
each stacking gel well, and perform the electrophoresis. The
Mobile phase A B C D molecular mass of the main electrophoretic band is between
Citric acid 12,500 and 13,800 when the band is stained by immersion in
monohydrate 17.70 g 10.50 g 6.10 g —
Coomassie staining TS.
Trisodium citrate
dihydrate 7.74 g 15.70 g 26.67 g — pH <2.54> 4.5 – 5.5
Sodium chloride 7.07 g 2.92 g 54.35 g —
Sodium hydroxide — — 2.30 g 8.00 g
Purity (1) Related substances—Perform the test with 10
Methanol (99.5) 40 mL — — — mL each of Celmoleukin (Genetical Recombination) and 0.01
Benzyl alcohol — 10 mL 5 mL — mol/L acetic acid buffer solution (pH 5.0) as directed under
Thiodiglycol 5 mL 5 mL 5 mL — Liquid Chromatography <2.01> under the following condi-
Lauromacrogol 4 mL 4 mL 4 mL 4 mL
tions, and measure the area of each peak by an automatic in-
solution (1 in 4) tegration method. When the amounts of related substances
Caprylic acid 0.1 mL 0.1 mL 0.1 mL 0.1 mL other than celmoleukin are calculated by the area percentage
Water a sufficient a sufficient a sufficient a sufficient
quantity quantity quantity quantity method, the total amount is not more than 5z.
Total 1000 mL 1000 mL 1000 mL 1000 mL Detector: An ultraviolet absorption photometer (wave-
length: 215 nm).
Flowing of mobile phase: Control the gradient by mixing Column: Stainless steel tube with an inside diameter of 4
the mobile phases A, B, C and D as directed in the following mm and a length of 30 cm packed with octadecylsilanized
table. silica gel for liquid chromatography (particle size: 5 mm).
259C.
Mobile phase A: A solution of trifluoroacetic acid in a
mixture of acetic acid and water (3:2) (1 in 1000).
Mobile phase B: A solution of trifluoroacetic acid in a
mixture of acetic acid and water (13:7) (1 in 1000).
670 Celmoleukin (Genetical Recombination) / Official Monographs JP XVII
Flowing of mobile phase: Control the gradient by mixing MS: Amount (mg) of ammonium chloride taken
the mobile phases A and B as directed in the following table. 0.003: Dilution correction factor
1.441: Molecular mass conversion coefficient for convert-
Time after injection Mobile phase A Mobile phase B ing ammonium chloride to ammonium acetate
of sample (min) (volz) (volz) Operating conditions—
Detector: An electric conductivity detector.
0 – 60 70 → 10 30 → 90
Column: Resin column 5 mm in inside diameter and 25 cm
in length, packed with weakly acidic ion exchange resin for
Flow rate: Adjust so that the retention time of celmoleu- liquid chromatography (particle size: 5.5 mm).
kin is about 50 minutes. Column temperature: A constant temperature of about
Time span of measurement: About 1.3 times as long as the 409C.
retention time of celmoleukin, beginning after the solvent Mobile phase: Diluted 0.1 mol/L methanesulfonic acid TS
peak. (3 in 10).
System suitability— Flow rate: Adjust so that the retention time of ammonium
Test for required detectability: Measure exactly 0.5 mL of is about 8 minutes.
Celmoleukin (Genetical Recombination), and add 0.01 System suitability—
mol/L acetic acid buffer solution (pH 5.0) to make exactly System performance: Measure exactly 1 mL of Standard
50 mL. Confirm that the peak area of celmoleukin obtained Sodium Stock Solution and 0.2 mL of Standard Potassium
from 10 mL of this solution is equivalent to 0.9 to 1.1z of Stock Solution, and then add water to make exactly 100 mL.
the peak area obtained from 10 mL of Celmoleukin (Geneti- Measure exactly 5 mL of this solution and 3 mL of Standard
cal Recombination). Ammonium Solution, and then add water to make exactly 50
System performance: Add 2 mL of 2-mercaptoethanol to mL. When 25 mL of this solution is run under the above con-
100 mL of Celmoleukin (Genetical Recombination), leave at ditions, sodium, ammonium and potassium are eluted in this
379 C for 2 hours, and then run this solution under the above order with the resolution between the peaks of sodium and
conditions. Celmoleukin and its reduced form are eluted in ammonium being not less than 3.0.
this order with the resolution between these peaks being not System repeatability: When the test is repeated 5 times
less than 3.0. with 25 mL of the standard solution under the above condi-
(2) Multimers—Dilute (at least 4 steps) the sample solutions, the relative standard deviation of the ammonium peak
tion prepared in the Molecular mass with buffer solution for area is not more than 10z.
celmoleukin so that the protein content is within the range of
2 to 32 mg per mL to prepare a series of standard solutions. Assay (1) Total protein content—Measure accurately 1
Pipet 20 mL each of the sample solution and the standard mL of Celmoleukin (Genetical Recombination) and add
solutions into the stacking gel well, and perform vertical water to make exactly 10 mL. Use this solution as the sample
uncoupled buffer SDS-polyacrylamide gel electrophoresis solution. Separately, weigh accurately about 50 mg of bovine
followed by immersion in Coomassie staining TS. Each elec- serum albumin for assay in water to prepare standard dilu-
trophoretic band is stained blue. Next, determine the peak tion solutions of 50, 100, and 150 mg/mL. Measure exactly 1
area of the electrophoretic bands obtained from each stand- mL of the sample solution and each standard dilution solu-
ard solution using a densitometer and calculate the protein tion, add exactly 2.5 mL of alkaline copper TS for protein
content using the calibration curve mentioned above. When content determination, shake, and leave for 15 minutes.
determining the polymer proteins derived from celmoleukin, Next, add exactly 2.5 mL of water and 0.5 mL of dilute
other than celmoleukin monomer, the amount is not more Folin's TS, and leave at 379 C for 30 minutes. Measure the
than 2z in relation to the total protein. absorbances of these solutions at 750 nm as directed under
(3) Host cell proteins—Being specified separately when Ultraviolet-visible Spectrophotometry <2.24>, using 1 mL of
the drug is granted approval based on the Law. water processed in the same way as control. Using the
(4) DNA—Being specified separately when the drug is calibration curve prepared from the absorbance of the stand-
granted approval based on the Law. ard dilution solution, calculate the protein content of Cel-
moleukin (Genetical Recombination).
Bacterial endotoxins <4.01> Less than 100 EU/mL. (2) Specific activity—Measure exactly 0.1 mL of Cel-
Ammonium acetate Measure exactly 0.1 mL of Celmoleu- moleukin (Genetical Recombination) and add exactly 0.9 mL
kin (Genetical Recombination), add water to make exactly of culture medium for celmoleukin to make the sample
10 mL, and use this solution as the sample solution. Sepa- solution. Separately, take one Interleukin-2 RS and add
rately, weigh accurately about 0.1 g of ammonium chloride, exactly 1 mL of water to dissolve. This is the standard solu-
and add water to make exactly 100 mL. Measure exactly 5 tion. Dilute exactly the sample and standard solutions in
mL of this solution, add water to make exactly 100 mL, and serially two-fold steps with culture medium for celmoleukin,
use this solution as the standard stock solution. Measure ex- and add equal volumes of interleukin-2 dependent mouse
actly 3 mL of the standard stock solution, add water to make natural killer NKC3 cells to the serially diluted solutions.
exactly 50 mL, and use this solution as the standard solution. The control solution is a mixture of equal volumes of inter-
Perform the test with exactly 25 mL each of the sample solu- leukin-2 dependent mouse natural killer NKC3 and culture
tion and standard solution as directed under Liquid Chroma- medium for celmoleukin. Incubate these solutions at 379 C
tography <2.01> according to the following conditions. When for 16 to 24 hours. Following this, add a volume of 3-(4,5-
determining the area of the ammonium ion peak AT and AS, dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazolium bromide
Celmoleukin (Genetical Recombination) contains not less TS that is 1/5 that of the volume of culture medium for cel-
than 0.28 mg and not more than 0.49 mg of ammonium ace- moleukin, incubate at 379C for 4 to 6 hours, add a volume
tate per mL. of sodium lauryl sulfate TS equivalent to the volume of the
culture medium for celmoleukin, and leave for 24 to 48
Amount (mg) of ammonium acetate (CH3COONH4) per mL hours. After eluting the blue-colored pigment generated,
＝ AT/AS × MS × 0.003 × 1.441 perform the test on these solutions as directed under Ultravi-
JP XVII Official Monographs / Cetirizine Hydrochloride 671
olet-visible Spectrophotometry <2.24>, and measure the ab-

sorbance at 590 nm. Taking the absorbance obtained when Cetirizine Hydrochloride
1000 to 2000 units of celmoleukin per mL are added as 100z
and the absorbance of the control solution as 0z, determine セチリジン塩酸塩
the dilution factor (A) of the Interleukin-2 RS that shows an
absorbance of 50z and dilution factor of Celmoleukin
(Genetical Recombination) (B). Multiply the B/A value by
the unit number of the Interleukin-2 RS to calculate the bio-
logical activity of 1 mL of Celmoleukin (Genetical Recombi-
nation). Calculate the ratio of biological activity in relation
to protein content determined in the total protein content
C21H25ClN2O3.2HCl: 461.81
test.
2-(2-{4-[(RS )-(4-Chlorophenyl)(phenyl)methyl]piperazin-
Containers and storage Containers—Tight containers. 1-yl}ethoxy)acetic acid dihydrochloride
Storage—At －209C or lower. [83881-52-1]
Cetirizine Hydrochloride, when dried, contains

Cetanol not less than 99.0z and not more than 101.0z of
cetirizine hydrochloride (C21H25ClN2O3.2HCl).
セタノール
Description Cetirizine Hydrochloride occurs as a white
crystalline powder.
Cetanol is a mixture of solid alcohols, and consists It is very soluble in water, and slightly soluble in ethanol
chiefly of cetanol (C16H34O: 242.44). (99.5).
Description Cetanol occurs as unctuous, white flakes,
A solution of Cetirizine Hydrochloride (1 in 10) shows no
granules, or masses. It has a faint, characteristic odor. It is
optical rotation.
tasteless.
It is very soluble in pyridine, freely soluble in ethanol (95), Identification (1) Determine the absorption spectrum of a
in ethanol (99.5) and in diethyl ether, very slightly soluble in solution of Cetirizine Hydrochloride in 0.1 mol/L hydro-
acetic anhydride, and practically insoluble in water. chloric acid TS (1 in 50,000) as directed under Ultraviolet-
Melting point <1.13> 47 – 539C Prepare the sample ac-
cording to Method 2, then attach tightly a capillary tube to
the bottom of the thermometer by means of a rubber band
or by any suitable means, and make the bottom of the capil-
Cetirizine Hydrochloride as directed in the potassium chlo-
lary tube equal in position to the lower end of the thermome-
ter. Insert this thermometer into a test tube 17 mm in inside
diameter and about 170 mm in height, fasten the thermome-
ter with cork stopper so that the lower end of the thermome-
same wave numbers.
ter is about 25 mm distant from the bottom of the test tube.
(3) A solution of Cetirizine Hydrochloride (1 in 100)
Suspend the test tube in a beaker containing water, and heat
the beaker with constant stirring until the temperature rises
to 59 C below the expected melting point. Then regulate the Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
rate of increase to 19C per minute. The temperature at which Cetirizine Hydrochloride according to Method 2, and per-
the sample is transparent and no turbidity is produced is form the test. Prepare the control solution with 2.0 mL of
taken as the melting point. Standard Lead Solution (not more than 10 ppm).
(2) Related substances—Dissolve 0.10 g of Cetirizine Hy-
Acid value <1.13> Not more than 1.0.
drochloride in 50 mL of the mobile phase, and use this solu-
Ester value <1.13> Not more than 2.0. tion as the sample solution. Pipet 2 mL of the sample solu-
tion, add the mobile phase to make exactly 50 mL. Pipet 5
Hydroxyl value <1.13> 210 – 232
mL of this solution, add the mobile phase to make exactly
Iodine value <1.13> Not more than 2.0. 100 mL, and use this solution as the standard solution. Per-
Purity (1) Clarity of solution—Dissolve 3.0 g of Cetanol
in 25 mL of ethanol (99.5) by warming: the solution is clear.
(2) Alkalinity—To the solution obtained in (1) add 2
termine each peak area of each solution by the automatic in-
drops of phenolphthalein TS: no red color develops.
tegration method: the area of each peak other than cetirizine
Residue on ignition <2.44> Not more than 0.05z (2 g). obtained from the sample solution is not larger than the peak
area of cetirizine obtained from the standard solution. And
the total area of the peaks other than cetirizine from the
ers.
sample solution is not larger than 2.5 times the peak area of
cetirizine from the standard solution.
length: 230 nm).
chromatography (5 mm in particle diameter).
672 Cetirizine Hydrochloride Tablets / Official Monographs JP XVII
Column temperature: A constant temperature of about Content uniformity test.
259 C. Take 1 tablet of Cetirizine Hydrochloride Tablets, add
Mobile phase: A mixture of acetonitrile and diluted 0.5 4V/5 mL of sodium 1-heptanesulfonate solution (1 in 5000)
mol/L sulfuric acid TS (2 in 25) (47:3). adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS, treat
Flow rate: Adjust so that the retention time of cetirizine is with ultrasonic waves for 20 minutes, add sodium 1-hep-
about 9 minutes. tanesulfonate solution (1 in 5000) adjusted to pH 3.0 with
Time span of measurement: About 3 times as long as the 0.5 mol/L sulfuric acid TS to exactly V mL so that each mL
retention time of cetirizine, beginning after the solvent peak. contains about 0.2 mg of cetirizine hydrochloride
System suitability— (C21H25ClN2O3.2HCl), and filter through a membrane filter
Test for required detectability: Pipet 5 mL of the standard with a pore size not exceeding 0.45 mm. Discard the first 3
solution, and add the mobile phase to make exactly 10 mL. mL of the filtrate, pipet 5 mL of the subsequent filtrate, add
Confirm that the peak area of cetirizine obtained from 10 mL exactly 2 mL of the internal standard solution, add aceto-
of this solution is equivalent to 35 to 65z of that obtained nitrile to make 10 mL, and use this solution as the sample so-
from 10 mL of the standard solution. lution. Then, proceed as directed in the Assay.
System performance: Dissolve 20 mg of Cetirizine Hydro-
Amount (mg) of cetirizine hydrochloride
chloride in the mobile phase to make 100 mL. To 5 mL of
(C21H25ClN2O3.2HCl)
this solution, add 3 mL of a solution of aminopyrine in the
＝ MS × QT/QS × V/100
mobile phase (1 in 2500), and add the mobile phase to make
20 mL. When the procedure is run with 10 mL of this solu- MS: Amount (mg) of cetirizine hydrochloride for assay
tion under the above operating conditions, cetirizine and taken
aminopyrine are eluted in this order with the resolution be-
with 10 mL of the standard solution under the above operat- Dissolution <6.10> When the test is performed at 50 revolu-
ing conditions, the relative standard deviation of the peak tions per minute according to the Paddle method, using 900
area of cetirizine is not more than 2.0z. mL of water as the dissolution medium, the dissolution rates
in 15 minutes of 5-mg tablet and in 30 minutes of 10-mg
um, 609C, 3 hours).
tively.
Residue on ignition <2.44> Not more than 0.2z (1 g). Start the test with 1 tablet of Cetirizine Hydrochloride
Assay Weigh accurately about 0.1 g of Cetirizine Hydro-
acetone and water (7:3), and titrate <2.50> to the second
equivalence point with 0.1 mol/L sodium hydroxide VS
(potentiometric titration). Perform a blank determination in
each mL contains about 5.6 mg of cetirizine hydrochloride
(C21H25ClN2O3.2HCl), and use this solution as the sample
Each mL of 0.1 mol/L sodium hydroxide VS solution. Separately, weigh accurately about 28 mg of cetiri-
＝ 15.39 mg of C21H25ClN2O3.2HCl zine hydrochloride for assay, previously dried in vacuum at
609C for 3 hours, and dissolve in water to make exactly 100
mL. Pipet 2 mL of this solution, add water to make exactly
ers.
100 mL, and use this solution as the standard solution.
Determine the absorbances, AT and AS, at 230 nm of the
sample solution and the standard solution as directed under
Cetirizine Hydrochloride Tablets Ultraviolet-visible Spectrophotometry <2.24>.
セチリジン塩酸塩錠 Dissolution rate (z) with respect to the labeled amount
of cetirizine hydrochloride (C21H25ClN2O3.2HCl)
＝ MS × AT/AS × V?/V × 1/C × 18
Cetirizine Hydrochloride Tablets contain not less
than 95.0z and not more than 105.0z of the MS: Amount (mg) of cetirizine hydrochloride for assay
labeled amount of cetirizine hydrochloride taken
(C21H25ClN2O3.2HCl: 461.81). C: Labeled amount (mg) of cetirizine hydrochloride
(C21H25ClN2O3.2HCl) in 1 tablet
with Cetirizine Hydrochloride. Assay Weigh accurately not less than 20 Cetirizine Hydro-
chloride Tablets, and powder. Weigh accurately a portion of
Identification To a quantity of powdered Cetirizine Hydro-
the powder, equivalent to about 10 mg of cetirizine hydro-
chloride Tablets, equivalent to 10 mg of Cetirizine Hydro-
chloride (C21H25ClN2O3.2HCl), add 40 mL of sodium 1-hep-
chloride, add about 70 mL of 0.1 mol/L hydrochloric acid
tanesulfonate solution (1 in 5000) adjusted to pH 3.0 with
TS, shake, add 0.1 mol/L hydrochloric acid TS to make 100
0.5 mol/L sulfuric acid TS, treat with ultrasonic waves for
mL, and filter. To 4 mL of the filtrate add 0.1 mol/L hydro-
20 minutes, add sodium 1-heptanesulfonate solution (1 in
chloric acid TS to make 25 mL, and determine the absorp-
5000) adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS, to
make exactly 50 mL, and filter this solution through a mem-
brane filter with a pore size not exceeding 0.45 mm. Discard
the first 3 mL of the filtrate, pipet 5 mL of the subsequent
Uniformity of dosage units <6.02> Perform the test accord- filtrate, add exactly 2 mL of the internal standard solution,
ing to the following method: it meets the requirement of the add acetonitrile to make exactly 10 mL, and use this solution
JP XVII Official Monographs / Cetotiamine Hydrochloride Hydrate 673
as the sample solution. Separately, weigh accurately about less than 98.0z and not more than 102.0z of cetotia-
20 mg of cetirizine hydrochloride for assay, previously dried mine hydrochloride (C18H26N4O6S.HCl: 462.95), cal-
in vacuum at 609C for 3 hours, and add sodium 1-hep- culated on the anhydrous basis.
tanesulfonate solution (1 in 5000) adjusted to pH 3.0 with
Description Cetotiamine Hydrochloride Hydrate occurs as
0.5 mol/L sulfuric acid TS, to make exactly 100 mL. Pipet 5
white, crystals or crystalline powder. It is odorless or has a
mL of this solution, add exactly 2 mL of the internal stand-
faint characteristic odor.
ard solution, add acetonitrile to make 10 mL, and use this
solution as the standard solution. Perform the test with 20
mL each of the sample solution and standard solution as di-
the following conditions, and calculate the ratios, QT and Identification (1) Determine the absorption spectrum of a
QS, of the peak area of cetirizine to that of the internal solution of Cetotiamine Hydrochloride Hydrate in 0.01
standard. mol/L hydrochloric acid TS (1 in 50,000) as directed under
Amount (mg) of cetirizine hydrochloride
(C21H25ClN2O3.2HCl)
of a solution of Cetotiamine Hydrochloride RS prepared in
＝ MS × QT/QS × 1/2
MS: Amount (mg) of cetirizine hydrochloride for assay similar intensities of absorption at the same wavelengths.
taken (2) Determine the infrared absorption spectrum of
Cetotiamine Hydrochloride Hydrate as directed in the potas-
Spectrum or the spectrum of Cetotiamine Hydrochloride
length: 230 nm).
(3) A solution of Cetotiamine Hydrochloride Hydrate (1
in 50) responds to the Qualitative Tests <1.09> for chloride.
Column temperature: A constant temperature of about Purity (1) Clarity and color of solution—A solution ob-
259 C. tained by dissolving 1.0 g of Cetotiamine Hydrochloride Hy-
Mobile phase: A mixture of a solution of sodium 1-hep- drate in 10 mL of water is clear and has no more color than
tansulfonate (1 in 2900) and acetonitrile (29:21), adjusted to the following control solution.
pH 3.0 with 0.5 mol/L sulfuric acid TS. Control solution: Mix exactly 1.5 mL of Cobalt (II) Chlo-
Flow rate: Adjust so that the retention time of cetirizine is ride CS, exactly 36 mL of Iron (III) Chloride CS and exactly
about 5 minutes. 12.5 mL of diluted dilute hydrochloric acid (1 in 10). Pipet 1
System suitability— mL of this mixture, and add diluted dilute hydrochloric acid
System performance: When the procedure is run with 20 (1 in 10) to make exactly 100 mL.
mL of the standard solution under the above operating con- (2) Heavy metals <1.07>—Proceed with 1.0 g of Cetotia-
ditions, cetirizine and the internal standard are eluted in this mine Hydrochloride Hydrate according to Method 1, and
order with the resolution between these peaks being not less perform the test. Prepare the control solution with 2.0 mL of
than 7. Standard Lead Solution (not more than 20 ppm).
System repeatability: When the test is repeated 6 times (3) Related substances—Dissolve 50 mg of Cetotiamine
with 20 mL of the standard solution under the above operat- Hydrochloride Hydrate in 50 mL of the mobile phase, and
ing conditions, the relative standard deviation of the ratio of use this solution as the sample solution. Pipet 1 mL of the
the peak area of cetirizine to that of the internal standard is sample solution, add the mobile phase to make exactly 200
not more than 1.0z. mL, and use this solution as the standard solution. Perform
ers.
area of the peak other than cetotiamine from the sample so-
Cetotiamine Hydrochloride Hydrate lution is not larger than the peak area of cetotiamine from
セトチアミン塩酸塩水和物
than cetotiamine from the sample solution is not larger than
2 times the peak area of cetotiamine from the standard solu-
tion.
the Assay.
C18H26N4O6S.HCl.H2O: 480.96
retention time of cetotiamine, beginning after the solvent
(3Z)-4-sN-[(4-Amino-2-methylpyrimidin-5-yl)methyl]-N-
peak.
formylaminot-3-(ethoxycarbonylsulfanyl)pent-3-enyl ethyl
carbonate monohydrochloride monohydrate
mL. Confirm that the peak area of cetotiamine obtained
Cetotiamine Hydrochloride Hydrate contains not
674 Cetraxate Hydrochloride / Official Monographs JP XVII
with 10 mL of this solution is equivalent to 7 to 13z of that
obtained with 10 mL of the standard solution. Cetraxate Hydrochloride
mL of the standard solution under the above operating con- セトラキサート塩酸塩
factor of the peak of cetotiamine are not less than 3000 and
0.7 – 1.0, respectively.
area of cetotiamine is not more than 2.0z. C17H23NO4.HCl: 341.83
3-{4-[trans-4-(Aminomethyl)cyclohexylcarbonyloxy]-
Water <2.48> 3.0 – 5.0z (40 mg, coulometric titration).
phenyl}propanoic acid monohydrochloride
Residue on ignition <2.44> Not more than 0.2z (1 g). [27724-96-5]
Assay Weigh accurately about 30 mg each of Cetotiamine
Cetraxate Hydrochloride, when dried, contains
Hydrochloride Hydrate and Cetotiamine Hydrochloride RS
not less than 98.5z of cetraxate hydrochloride
(C17H23NO4.HCl).
Cetotiamine Hydrochloride Hydrate), add exactly 10 mL
each of the internal standard solution, then add a mixture of Description Cetraxate Hydrochloride occurs as white, crys-
water and methanol (1:1) to make 50 mL. To 2 mL each of tals or crystalline powder.
these solutions add a mixture of water and methanol (1:1) to It is soluble in methanol, sparingly soluble in water and in
make 10 mL, and use these solutions as the sample solution ethanol (95), and practically insoluble in diethyl ether.
and the standard solution, respectively. Perform the test Melting point: about 2369 C (with decomposition).
solution of Cetraxate Hydrochloride in methanol (1 in 2500)
QS, of the peak area of cetotiamine to that of the internal
standard.
Amount (mg) of cetotiamine hydrochloride
(C18H26N4O6S.HCl)
(2) Dissolve 0.5 g of Cetraxate Hydrochloride in 5 mL of
＝ MS × QT/QS
a mixture of water and 2-propanol (1:1) by warming, cool to
MS: Amount (mg) of Cetotiamine Hydrochloride RS below 259C. Filter, dry the formed crystals in vacuum for 4
taken, calculated on the anhydrous basis hours, and further dry at 1059C for 1 hour. Determine the
infrared absorption spectrum of the dried matter as directed
in the potassium chloride disk method under Infrared Spec-
droxybenzoate in a mixture of water and methanol (1:1) (1 in
800).
Reference Spectrum: both spectra exhibit similar intensities
(3) A solution of Cetraxate Hydrochloride (1 in 100)
length: 245 nm).
responds to the Qualitative Tests <1.09> (2) for chloride.
ter and 15 cm in length, packed with octadecylsilanized silica Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
gel for liquid chromatography (5 mm in particle diameter). Cetraxate Hydrochloride according to Method 2, and per-
Column temperature: A constant temperature of about form the test. Prepare the control solution with 2.0 mL of
259 C. Standard Lead Solution (not more than 10 ppm).
Mobile phase: Dissolve 1.0 g of sodium 1-heptanesul- (2) Arsenic <1.11>—Prepare the test solution with 1.0 g
fonate in diluted acetic acid (100) (1 in 100) to make 1000 of Cetraxate Hydrochloride according to Method 3, and per-
mL. To 1 volume of this solution add 1 volume of methanol. form the test with a solution of magnesium nitrate hexahy-
Flow rate: Adjust so that the retention time of cetotiamine drate in ethanol (95) (1 in 5) (not more than 2 ppm).
is about 10 minutes. (3) cis Isomer—Dissolve 0.10 g of Cetraxate Hydrochlo-
System suitability— ride in 10 mL of water, and use this solution as the sample
System performance: When the procedure is run with 5 mL solution. To exactly 5 mL of the sample solution add water
of the standard solution under the above operating condi- to make exactly 100 mL. To exactly 2 mL of this solution
tions, cetotiamine and the internal standard are eluted in this add water to make exactly 50 mL, and use this solution as
order with the resolution between these peaks being not less the standard solution. Perform the test with exactly 10 mL
than 5. each of the sample solution and standard solution as directed
System repeatability: When the test is repeated 6 times under Liquid Chromatography <2.01> according to the fol-
with 5 mL of the standard solution under the above operating lowing conditions. Determine each peak area of both solu-
conditions, the relative standard deviation of the ratio of the tions by the automatic integration method: the area of the
peak area of cetotiamine to that of the internal standard is peak, having the relative retention time 1.3 to 1.6 to
not more than 1.0z. cetraxate from the sample solution is not larger than the
peak area of cetraxate from the standard solution.
length: 220 nm).
JP XVII Official Monographs / Chenodeoxycholic Acid 675
and 15 cm in length, packed with octadecylsilanized silica gel mL each of the sample solution and standard solution on a
for liquid chromatography (5 mm in particle diameter). plate of silica gel for thin-layer chromatography. Develop
Column temperature: A constant temperature of about the plate with a mixture of chloroform, methanol and acetic
259 C. acid (100) (20:4:3) to a distance of about 10 cm, and air-dry
Mobile phase: Adjust the pH of a mixture of water, meth- the plate. Spray evenly ninhydrin TS on the plate, and heat
anol and 0.5 mol/L ammonium acetate TS (15:10:4) to 6.0 at 909C for 10 minutes: the spots other than the principal
with acetic acid (31). spot from the sample solution are not more intense than the
Flow rate: Adjust so that the retention time of cetraxate is spot from the standard solution.
about 10 minutes.
3 hours).
System performance: Dissolve 0.02 g of Cetraxate Hydro-
chloride and 0.01 g of phenol in 100 mL of water. To 2 mL Residue on ignition <2.44> Not more than 0.1z (1 g).
of this solution add water to make 20 mL. When the proce-
Assay Weigh accurately about 0.5 g of Cetraxate Hydro-
dure is run with 10 mL of this solution under the above oper-
chloride, previously dried, dissolve in 100 mL of water, and
ating conditions, cetraxate and phenol are eluted in this
adjust the pH of this solution to between 7.0 and 7.5 with
dilute sodium hydroxide TS. To this solution add 10 mL of
than 5.
formaldehyde solution, stir for about 5 minutes, and titrate
<2.50> with 0.1 mol/L sodium hydroxide VS by taking over
about 20 minutes (potentiometric titration). Perform a blank
area of cetraxate is not more than 2.0z.
(4) 3-( p-Hydroxyphenyl)propionic acid—To 0.10 g of Each mL of 0.1 mol/L sodium hydroxide VS
Cetraxate Hydrochloride add exactly 2 mL of the internal ＝ 34.18 mg of C17H23NO4.HCl
standard solution and methanol to make 10 mL, and use this
of 3-( p-hydroxyphenyl)propionic acid in methanol to make
exactly 100 mL. To exactly 2 mL of this solution add exactly
2 mL of the internal standard solution and methanol to Chenodeoxycholic Acid
ケノデオキシコール酸
the ratios, QT and QS, of the peak area of 3-( p-hydroxy-
phenyl)propionic acid to that of the internal standard: QT is
not larger than QS.
Internal standard solution—A solution of caffeine in metha-
nol (1 in 4000).
C24H40O4: 392.57
3a,7a-Dihydroxy-5b-cholan-24-oic acid
length: 230 nm).
[474-25-9]
Chenodeoxycholic Acid, when dried, contains not
chenodeoxycholic acid (C24H40O4).
409 C.
Mobile phase: Adjust the pH of a mixture of water, meth- Description Chenodeoxycholic Acid occurs as white, crys-
anol and 0.5 mol/L ammonium acetate TS (15:5:2) to 5.5 tals, crystalline powder or powder.
with acetic acid (31). It is freely soluble in methanol and in ethanol (99.5), solu-
Flow rate: Adjust so that the retention time of 3-( p- ble in acetone, and practically insoluble in water.
hydroxyphenyl)propionic acid is about 7 minutes.
of Chenodeoxycholic Acid, previously dried, as directed in
ditions, 3-(p-hydroxyphenyl)propionic acid and the internal
standard are eluted in this order with the resolution between
System repeatability: When the test is repeated 6 times Optical rotation <2.49> [a]20 D : ＋11.0 – ＋13.09(after dry-
with 10 mL of the standard solution under the above operating, 0.4 g, ethanol (99.5), 20 mL, 100 mm).
the peak area of 3-( p-hydroxyphenyl)propionic acid to that
of the internal standard is not more than 1.0z. Purity (1) Chloride <1.03>—Dissolve 0.36 g of
(5) Related substances—Dissolve 0.10 g of Cetraxate Hy- Chenodeoxycholic Acid in 30 mL of methanol, add 10 mL of
drochloride in 10 mL of methanol, and use this solution as dilute nitric acid and water to make 50 mL, and perform the
the sample solution. Pipet 1 mL of the sample solution, add test with this solution. Prepare the control solution as
methanol to make exactly 100 mL, and use this solution as follows: To 1.0 mL of 0.01 mol/L hydrochloric acid VS add
the standard solution. Perform the test with these solutions 30 mL of methanol, 10 mL of dilute nitric acid and water to
as directed under Thin-layer Chromatography <2.03>. Spot 5 make 50 mL (not more than 0.1z).
676 Chloral Hydrate / Official Monographs JP XVII
Chenodeoxycholic Acid according to Method 4, and per- Chloral Hydrate
Standard Lead Solution (not more than 20 ppm). 抱水クロラール
(3) Barium—To 2.0 g of Chenodeoxycholic Acid add
100 mL of water, and boil for 2 minutes. To this solution
add 2 mL of hydrochloric acid, boil for 2 minutes, filter
after cooling, and wash the filter with water until to get 100
mL of the filtrate. To 10 mL of the filtrate add 1 mL of
C2H3Cl3O2: 165.40
dilute sulfuric acid: no turbidity appears.
2,2,2-Trichloroethane-1,1-diol
(4) Related substances—Dissolve 0.20 g of Chenodeoxy-
[302-17-0]
cholic Acid in a mixture of acetone and water (9:1) to make
Chloral Hydrate contains not less than 99.5z of
Separately, dissolve 10 mg of lithocholic acid for thin-layer
chloral hydrate (C2H3Cl3O2).
chromatography in the mixture of acetone and water (9:1) to
make exactly 10 mL. Pipet 2 mL of this solution, add the Description Chloral Hydrate occurs as colorless crystals. It
mixture of acetone and water (9:1) to make exactly 100 mL, has a pungent odor and an acrid, slightly bitter taste.
and use this solution as the standard solution (1). Separately, It is very soluble in water, and freely soluble in ethanol
dissolve 10 mg of ursodeoxycholic acid in the mixture of ace- (95) and in diethyl ether.
tone and water (9:1) to make exactly 100 mL, and use this It slowly volatilizes in air.
solution as the standard solution (2). Separately, dissolve 10
Identification (1) Dissolve 0.2 g of Chloral Hydrate in 2
mg of cholic acid for thin-layer chromatography in the mix-
mL of water, and add 2 mL of sodium hydroxide TS: the
ture of acetone and water (9:1) to make exactly 100 mL, and
turbidity is produced, and it separates into two clear layers
use this solution as the standard solution (3). Pipet 1 mL of
by warming.
the sample solution, and add the mixture of acetone and
(2) Heat 0.2 g of Chloral Hydrate with 3 drops of aniline
water (9:1) to make exactly 20 mL. Pipet 0.5 mL, 1 mL, 2
and 3 drops of sodium hydroxide TS: the disagreeable odor
mL, 3 mL and 5 mL of this solution, add the mixture of ace-
of phenylisocyanide (poisonous) is perceptible.
tone and water (9:1) to each of them to make exactly 50 mL,
and designate these solutions as standard solution A, stand- Purity (1) Clarity and color of solution—Dissolve 1.0 g
ard solution B, standard solution C, standard solution D and of Chloral Hydrate in 2 mL of water: the solution is clear
standard solution E, respectively. Perform the test with these and colorless.
solutions as directed under Thin-layer Chromatography (2) Acidity—Dissolve 0.20 g of Chloral Hydrate in 2 mL
<2.03>. Spot 5 mL each of the sample solution, standard solu- of water, and add 1 drop of methyl orange TS: a yellow
tions (1), (2), (3) and standard solutions A, B, C, D and E on color develops.
a plate of silica gel for thin-layer chromatography. Develop (3) Chloride <1.03>—Perform the test with 1.0 g of Chlo-
the plate with a mixture of 4-methyl-2-pentanone, toluene ral Hydrate. Prepare the control solution with 0.30 mL of
and formic acid (16:6:1) to a distance of about 15 cm, air- 0.01 mol/L hydrochloric acid VS (not more than 0.011z).
dry the plate, and further dry at 1209C for 30 minutes. Im- (4) Chloral alcoholate—Warm 1.0 g of Chloral Hydrate
mediately, spray evenly a solution of phosphomolybdic acid with 10 mL of sodium hydroxide TS, filter the upper layer,
n-hydrate in ethanol (95) (1 in 5) on the plate, and heat at add iodine TS to the filtrate until a yellow color develops,
1209C for 2 to 3 minutes: the spot corresponding to the spot and allow the solution to stand for 1 hour: no yellow precipi-
with the standard solution (1) is not more intense than the tate is produced.
spot with the standard solution (1), the spot corresponding (5) Benzene—Warm the solution obtained in (1) with 3
to the spot with the standard solution (2) is not more intense mL of water: no odor of benzene is perceptible.
than the spot with the standard solution (2), and the spot
corresponding to the spot with the standard solution (3) is
not more intense than the spot with the standard solution Assay Weigh accurately about 4 g of Chloral Hydrate in a
(3). As compared to the spots with the standard solutions A, glass-stoppered flask, add 10 mL of water and exactly 40 mL
B, C, D and E, the spots other than the principal spot and of 1 mol/L sodium hydroxide VS, and allow the mixture to
the spots mentioned above with the sample solution are not stand for exactly 2 minutes. Titrate <2.50> the excess sodium
more intense than the spot with the standard solution E, and hydroxide immediately with 0.5 mol/L sulfuric acid VS (in-
the total amount of them is not more than 1.5z. dicator: 2 drops of phenolphthalein TS). Perform a blank
3 hours). Each mL of 1 mol/L sodium hydroxide VS
＝ 165.4 mg of C2H3Cl3O2
Assay Weigh accurately about 0.5 g of Chenodeoxycholic
Acid, previously dried, dissolve in 40 mL of ethanol (95) and
20 mL of water, and titrate <2.50> with 0.1 mol/L sodium
hydroxide VS (potentiometric titration). Perform a blank
determination in the same manner, and make any necessary
correction.
＝ 39.26 mg of C24H40O4
JP XVII Official Monographs / Chloramphenicol Palmitate 677
from the sample solution is not more than 2.0z.

Chloramphenicol Loss on drying <2.41> Not more than 0.5z (1 g, 1059C,
3 hours).
クロラムフェニコール
Assay Weigh accurately an amount of Chloramphenicol
and Chloramphenicol RS, equivalent to about 0.1 g (po-
tency), dissolve each in 20 mL of methanol, and add water to
make exactly 100 mL. Pipet 20 mL each of these solutions,
C11H12Cl2N2O5: 323.13 and add water to make exactly 100 mL. Pipet 10 mL each of
2,2-Dichloro-N-[(1R,2R)-1,3-dihydroxy-1- these solutions, add water to make exactly 100 mL, and use
(4-nitrophenyl)propan-2-yl]acetamide these solutions as the sample solution and standard solution.
[56-75-7] Determine the absorbances, AT and AS, at 278 nm of the
Chloramphenicol contains not less than 980 mg (po- Ultraviolet-visible Spectrophotometry <2.24>.
tency) and not more than 1020 mg (potency) per mg,
Amount [ mg (potency)] of chloramphenicol (C11H12Cl2N2O5)
calculated on the dried basis. The potency of Chlo- ＝ MS × AT/AS × 1000
ramphenicol is expressed as mass (potency) of chlo-
ramphenicol (C11H12Cl2N2O5). MS: Amount [mg (potency)] of Chloramphenicol RS
taken
Description Chloramphenicol occurs as white to yellowish
white, crystals or crystalline powder. Containers and storage Containers—Tight containers.
It is freely soluble in methanol and in ethanol (99.5), and
slightly soluble in water.
Identification (1) Determine the absorption spectrum of Chloramphenicol Palmitate
クロラムフェニコールパルミチン酸エステル
of a solution of Chloramphenicol RS prepared in the same
Chloramphenicol as directed in the potassium bromide disk
C27H42Cl2N2O6: 561.54
(2R,3R)-2-(Dichloroacetyl)amino-3-hydroxy-3-
trum of Chloramphenicol RS: both spectra exhibit similar
(4-nitrophenyl)propan-1-yl palmitate
[530-43-8]
Optical rotation <2.49> [a]20D : ＋18.5 – ＋21.59 (1.25 g,
ethanol (99.5), 25 mL, 100 mm). Chloramphenicol Palmitate contains not less than
per mg, calculated on the dried basis. The potency of
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Chloramphenicol Palmitate is expressed as mass (po-
Chloramphenicol according to Method 2, and perform the tency) of chloramphenicol (C11H12Cl2N2O5: 323.13).
Description Chloramphenicol Palmitate occurs as a white
to grayish white, crystalline powder.
It is freely soluble in acetone, sparingly soluble in metha-
of Chloramphenicol according to Method 4, and perform
nol and in ethanol (99.5), and practically insoluble in water.
(3) Related substances—Dissolve 0.10 g of Chloram- Identification (1) Determine the absorption spectrum of a
phenicol in 10 mL of methanol, and use this solution as the solution of Chloramphenicol Palmitate in ethanol (99.5) (1
sample solution. Pipet 1 mL of the sample solution, add in 33,000) as directed under Ultraviolet-visible Spectropho-
methanol to make exactly 100 mL, and use this solution as tometry <2.24>, and compare the spectrum with the Refer-
the standard solution (1). Pipet 10 mL of the standard solu- ence Spectrum or the spectrum of a solution of Chloram-
tion (1), add methanol to make exactly 20 mL, and use this phenicol Palmitate RS prepared in the same manner as the
solution as the standard solution (2). Perform the test with sample solution: both spectra exhibit similar intensities of
these solutions as directed under Thin-layer Chromatogra- absorption at the same wavelengths.
phy <2.03>. Spot 20 mL each of the sample solution and (2) Dissolve 5 mg each of Chloramphenicol Palmitate
standard solutions (1) and (2) on a plate of silica gel with and Chloramphenicol Palmitate RS in 1 mL of acetone, and
fluorescent indicator for thin-layer chromatography, de- use these solutions as the sample solution and standard sou-
velop the plate with a mixture of chloroform, methanol and tion. Perform the test with these solutions as directed under
acetic acid (100) (79:14:7) to a distance of about 15 cm, and Thin-layer Chromatography <2.03>. Spot 5 mL each of the
air-dry the plate. Examine under ultraviolet light (main sample solution and standard solution on a plate of silica gel
wavelength: 254 nm): the spots other than the principal spot with fluorescent indicator for thin-layer chromatography.
and the spot on the original obtained from the sample solu- Develop the plate with a mixture of acetone and cyclohexane
tion are not more intense than the spot obtained from the (1:1) to a distance of about 10 cm, and air-dry the plate.
standard solution (1), and the total amount of these spots Examine under ultraviolet light (main wavelength: 254 nm):
678 Chloramphenicol Sodium Succinate / Official Monographs JP XVII
the principal spot obtained from the sample solution has the Loss on drying <2.41> Not more than 1.0z (1 g, reduced
same R f value as the spot obtained from the standard solu- C, 3 hours).
pressure not exceeding 0.67 kPa, 609
tion.
Assay Weigh accurately an amount of Chloramphenicol
Optical rotation <2.49> [a]25D : ＋21 – ＋259(1 g calculated Palmitate and Chloramphenicol Palmitate RS, equivalent to
on the dried basis, ethanol (99.5), 20 mL, 100 mm). about 37 mg (potency), dissolve each in 40 mL of methanol
and 1 mL of acetic acid (100), and add methanol to make ex-
Melting point <2.60> 91 – 969
C
actly 50 mL. Pipet 10 mL each of these solutions, add the
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of mobile phase to make exactly 25 mL, and use these solutions
Chloramphenicol Palmitate according to Method 4, and as the sample solution and standard solution. Perform the
perform the test. Prepare the control solution with 2.0 mL of test with exactly 10 mL each of the sample solution and
Standard Lead Solution (not more than 20 ppm). standard solution as directed under Liquid Chromatography
(2) Arsenic <1.11>—Prepare the test solution with 1.0 g <2.01> according to the following conditions, and determine
of Chloramphenicol Palmitate according to Method 3, and the peak areas, AT and AS, of chloramphenicol palmitate in
perform the test (not more than 2 ppm). each solution.
(3) Related substances—Dissolve 50 mg of Chloram-
Amount [ mg (potency)] of chloramphenicol (C11H12Cl2N2O5)
phenicol Palmitate in 50 mL of methanol, and use this solu-
＝ MS × AT/AS × 1000
tion, add methanol to make exactly 100 mL, and use this MS: Amount [mg (potency)] of Chloramphenicol
solution as the standard solution. Perform the test with Palmitate RS taken
according to the following conditions. The test should be
length: 280 nm).
performed within 30 minutes after the sample solution and
standard solution are prepared. Determine each peak area by
the automatic integration method: the total area of the peaks
other than the peak of chloramphenicol palmitate from the
sample solution is not larger than 3.5 times the peak area of
409C.
chloramphenicol palmitate from the standard solution. For
Mobile phase: A mixture of methanol, water and acetic
the peak areas for chloramphenicol, having the relative
acid (100) (172:27:1).
retention time of about 0.5 to chloramphenicol palmitate,
Flow rate: Adjust so that the retention time of chloram-
and for chloramphenicol dipalmitate, having the relative
phenicol palmitate is about 7 minutes.
retention time of about 5.0 to chloramphenicol palmitate,
multiply their relative response factors, 0.5 and 1.4, respec-
tively.
ditions, the number of theoretical plates of the peak of chlo-
ramphenicol palmitate is not less than 2400.
length: 270 nm).
area of chloramphenicol palmitate is not more than 1.0z.
209 C. Containers and storage Containers—Tight containers.
Mobile phase: Methanol. Storage—Light-resistant.
Flow rate: Adjust so that the retention time of chloram-
phenicol palmitate is about 5 minutes.
Time span of measurement: About 6 times as long as the Chloramphenicol Sodium Succinate
retention time of chloramphenicol palmitate.
System suitability— クロラムフェニコールコハク酸エステルナトリウム
Test for required detectability: Dissolve 50 mg of Chlo-
ramphenicol Palmitate in 50 mL of methanol. To 1 mL of
this solution, add methanol to make 100 mL, and use this so-
lution as the solution for system suitability test. Pipet 5 mL
of the solution for system suitabillity test, and add methanol
to make exactly 50 mL. Confirm that the peak area of chlo-
ramphenicol palmitate obtained from 20 mL of this solution
is equivalent to 7 to 13z of that obtained from 20 mL of the C15H15Cl2N2NaO8: 445.18
solution for system suitability test. Monosodium (2R,3R)-2-(dichloroacetyl)amino-3-hydroxy-
System performance: When the procedure is run with 20 3-(4-nitrophenyl)propan-1-yl succinate
mL of the solution for system suitability test under the above [982-57-0]
operating conditions, the number of theoretical plates of the
peak of chloramphenicol palmitate is not less than 5000. Chloramphenicol Sodium Succinate contains not
System repeatability: When the test is repeated 6 times less than 711 mg (potency) and not more than 740 mg
with 20 mL of the solution for system suitability test under (potency) per mg, calculated on the anhydrous basis.
the above operating conditions, the relative standard devia- The potency of Chloramphenicol Sodium Succinate is
tion of the peak area of chloramphenicol palmitate is not expressed as mass (potency) of chloramphenicol
more than 1.0z. (C11H12Cl2N2O5: 323.13).
JP XVII Official Monographs / Chlordiazepoxide 679
Description Chloramphenicol Sodium Succinate occurs as

white to yellowish white, crystals or crystalline powder. Chlordiazepoxide
It is very soluble in water, and freely soluble in methanol
and in ethanol (99.5). クロルジアゼポキシド
It is hygroscopic.
solution of Chloramphenicol Sodium Succinate (1 in 50,000)
(2) Determine the infrared absorption spectrum of Chlo-
C16H14ClN3O: 299.75
ramphenicol Sodium Succinate as directed in the potassium
7-Chloro-2-methylamino-5-phenyl-3H-1,4-benzodiazepin-
4-oxide
[58-25-3]
Chlordiazepoxide, when dried, contains not less
(3) Chloramphenicol Sodium Succinate responds to the
than 98.5z of chlordiazepoxide (C16H14ClN3O).
Qualitative Tests <1.09> (1) for sodium salt.
Description Chlordiazepoxide occurs as white to light yel-
D : ＋5 – ＋89(1.25 g calculated
low, crystals or crystalline powder.
pH <2.54> The pH of a solution obtained by dissolving ethanol (95), very slightly soluble in diethyl ether, and practi-
1.4 g of Chloramphenicol Sodium Succinate in 5 mL of cally insoluble in water.
water is between 6.0 and 7.0. It dissolves in dilute hydrochloric acid.
It is gradually affected by light.
of Chloramphenicol Sodium Succinate in 10 mL of water:
the solution is clear and colorless to yellowish. Identification (1) Determine the absorption spectrum of a
(2) Heavy metals <1.07>—Proceed with 1.0 g of Chlo- solution of Chlordiazepoxide in 0.1 mol/L hydrochloric acid
ramphenicol Sodium Succinate according to Method 2, and TS (1 in 200,000) as directed under Ultraviolet-visible Spec-
perform the test. Prepare the control solution with 2.0 mL of trophotometry <2.24>, and compare the spectrum with the
Standard Lead Solution (not more than 20 ppm). Reference Spectrum or the spectrum of a solution of Chlor-
(3) Arsenic <1.11>—Prepare the test solution with 1.0 g diazepoxide RS prepared in the same manner as the sample
of Chloramphenicol Sodium Succinate according to Method solution: both spectra exhibit similar intensities of absorp-
1, and perform the test (not more than 2 ppm). tion at the same wavelengths.
(2) Determine the infrared absorption spectra of Chlor-
diazepoxide, previously dried, as directed in the potassium
Assay Weigh accurately an amount of Chloramphenicol <2.25>, and compare the spectrum with the Reference Spec-
Sodium Succinate, equivalent to about 20 mg (potency), dis- trum or the spectrum of dried Chlordiazepoxide RS: both
solve in water to make exactly 1000 mL, and use this solu- spectra exhibit similar intensities of absorption at the same
tion as the sample solution. Separately, weigh accurately an wave numbers.
amount of Chloramphenicol Succinate RS, equivalent to (3) Proceed with Chlordiazepoxide as directed under
about 20 mg (potency), add about 50 mL of water to make a Flame Coloration Test <1.04> (2), and perform the test: a
suspension, and add gradually about 7 mL of 0.01 mol/L so- green color develops.
dium hydroxide TS while stirring to adjust the pH to 7.0. To
this solution add water to make exactly 1000 mL, and use
Chlordiazepoxide according to Method 2, and perform the
this solution as the standard solution. Determine the absor-
bances, AT and AS, at 276 nm of the sample solution and
exposure to light, using light-resistant vessels. Dissolve
Amount [ mg (potency)] of chloramphenicol (C11H12Cl2N2O5) 0.20 g of Chlordiazepoxide in exactly 10 mL of a mixture of
＝ MS × AT/AS × 1000 methanol and ammonia TS (97:3), and use this solution as
MS: Amount [mg (potency)] of Chloramphenicol Suc-
a mixture of methanol and ammonia TS (97:3) to make ex-
cinate RS taken
Containers and storage Containers—Hermetic containers. (1). Separately, dissolve 10 mg of 2-amino-5-chloroben-
zophenone for thin-layer chromatography in methanol to
solution (2). Perform the test with these solutions as directed
under Thin-layer Chromatography <2.03>. Spot 25 mL of the
sample solution and 5 mL each of the standard solutions (1)
and (2) on a plate of silica gel with fluorescent indicator for
of ethyl acetate and ethanol (99.5) (19:1) to a distance of
680 Chlordiazepoxide Powder / Official Monographs JP XVII
about 12 cm, and air-dry the plate. Examine under ultravio- standard solution (1). Dissolve 5.0 mg of 2-amino-5-
let light (main wavelength: 254 nm): the spots other than the chlorobenzophenone for thin-layer chromatography in
principal spot from the sample solution are not more intense methanol to make exactly 200 mL, and use this solution as
than the spot from the standard solution (1). Spray evenly a the standard solution (2). Perform the test with these solu-
solution of sodium nitrite in 1 mol/L hydrochloric acid TS tions as directed under Thin-layer Chromatography <2.03>.
(1 in 100) on the plate, allow to stand for 1 minute, and Spot 25 mL of the sample solution and 10 mL each of the
spray evenly N-(1-naphthyl)-N?-diethylethylenediamine standard solutions (1) and (2) on a plate of silica gel with
oxalate-acetone TS on the plate: the spots from the sample fluorescent indicator for thin-layer chromatography. Pro-
solution are not more intense than the spots from the stand- ceed as directed in the Purity (2) under Chlordiazepoxide.
ard solution (2).
Loss on drying <2.41> Not more than 0.5z (1 g, in vacu- lutions per minute according to the Paddle method, using
um, phosphorus (V) oxide, 609C, 4 hours). 900 mL of 2nd fluid for dissolution test as the dissolution
medium, the dissolution rate in 60 minutes of Chlordia-
zepoxide Powder is not less than 70z.
Assay Weigh accurately about 0.6 g of Chlordiazepoxide, Start the test with an accurately weighed amount of Chlor-
previously dried, and dissolve in 50 mL of acetic acid (100). diazepoxide Powder, equivalent to about 3.3 mg of chlordia-
Titrate <2.50> with 0.1 mol/L perchloric acid VS until the zepoxide (C16H14ClN3O), withdraw not less than 15 mL of
color of the supernatant liquid changes from purple through the medium at the specified minute after starting the test,
blue-purple to blue (indicator: 3 drops of crystal violet TS). and filter through a membrane filter with a pore size not
Perform a blank determination in the same manner, and exceeding 0.45 mm. Discard the first 10 mL of the filtrate,
make any necessary correction. and use the subsequent filtrate as the sample solution. Sepa-
rately, weigh accurately about 12 mg of Chlordiazepoxide
RS, previously dried in a desiccator (in vacuum, phosphorus
＝ 29.98 mg of C16H14ClN3O
(V) oxide, 609C) for 4 hours, dissolve in 20 mL of 0.1 mol/L
Containers and storage Containers—Tight containers. hydrochloric acid TS, and add the dissolution medium to
Storage—Light-resistant. make exactly 200 mL. Pipet 3 mL of this solution, add the
Chlordiazepoxide Powder AT and AS, at 260 nm of the sample solution and standard
クロルジアゼポキシド散 tometry <2.24>.
Chlordiazepoxide Powder contains not less than of chlordiazepoxide (C16H14ClN3O)
93.0z and not more than 107.0z of the labeled ＝ MS/MT × AT/AS × 1/C × 27
amount of chlordiazepoxide (C16H14ClN3O: 299.75).
MS: Amount (mg) of Chlordiazepoxide RS taken
Method of preparation Prepare as directed under Granules MT: Amount (g) of Chlordiazepoxide Powder taken
or Powders, with Chlordiazepoxide. C: Labeled amount (mg) of chlordiazepoxide
(C16H14ClN3O) in 1 g
Identification (1) Weigh a portion of Chlordiazepoxide
Powder, equivalent to 0.01 g of Chlordiazepoxide, add 100 Assay Conduct this procedure without exposure to light,
mL of 0.1 mol/L hydrochloric acid TS, shake, and filter. To using light-resistant vessels. Weigh accurately a quantity
5 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to of Chlordiazepoxide Powder, equivalent to about 0.1 g
make 100 mL, and determine the absorption spectrum of of chlordiazepoxide (C16H14ClN3O), transfer to a glass-
this solution as directed under Ultraviolet-visible Spectro- stoppered flask, wet with exactly 10 mL of water, add ex-
photometry <2.24>: it exhibits maxima between 244 nm and actly 90 mL of methanol, stopper, shake vigorously for 15
248 nm and between 306 nm and 310 nm, and a minimum minutes, and centrifuge. Pipet 10 mL of the supernatant liq-
between 288 nm and 292 nm. uid, add exactly 5 mL of the internal standard solution, add
(2) Weigh a portion of Chlordiazepoxide Powder, methanol to make 100 mL, and use this solution as the sam-
equivalent to 0.02 g of Chlordiazepoxide, add 10 mL of ple solution. Separately, weigh accurately about 0.1 g of
methanol, shake for 5 minutes, then filter by suction Chlordiazepoxide RS, previously dried in a desiccator (in
through a glass filter (G4), evaporate the filtrate with the aid vacuum, phosphorus (V) oxide, 609C) for 4 hours, and dis-
of a current of air to dryness, and dry the residue in vacuum solve in exactly 10 mL of water and exactly 90 mL of metha-
at 609C for 1 hour. Determine the infrared absorption spec- nol. Pipet 10 mL of this solution, add exactly 5 mL of the in-
trum of the residue as directed in the potassium bromide disk ternal standard solution, add methanol to make 100 mL, and
method under Infrared Spectrophotometry <2.25>: it exhibits use this solution as the standard solution. Perform the test
absorption at the wave numbers of about 1625 cm－1, 1465 with 10 mL each of the sample solution and standard solution
cm－1, 1265 cm－1, 850 cm－1 and 765 cm－1. as directed under Liquid Chromatography <2.01> according
Purity Conduct this procedure without exposure to light,
QS, of the peak area of chlordiazepoxide to that of the inter-
using light-resistant vessels. To a portion of Chlordia-
nal standard.
zepoxide Powder, equivalent to 50 mg of Chlordiazepoxide,
add exactly 5 mL of a mixture of methanol and ammonia TS Amount (mg) of chlordiazepoxide (C16H14ClN3O)
(97:3), shake, centrifuge, and use the supernatant liquid as ＝ MS × QT/QS
the sample solution. Separately, dissolve 50 mg of Chlordia-
MS: Amount (mg) of Chlordiazepoxide RS taken
zepoxide RS in a mixture of methanol and ammonia TS
(97:3) to make exactly 50 mL, and use this solution as the Internal standard solution—A solution of isobutyl salicylate
JP XVII Official Monographs / Chlordiazepoxide Tablets 681
in methanol (1 in 20). 2-amino-5-chlorobenzophenone for thin-layer chromatogra-

Operating conditions— phy in methanol to make exactly 200 mL, and use this solu-
Detector: An ultraviolet absorption photometer (wave- tion as the standard solution (2). Perform the test with these
length: 254 nm). solutions as directed under Thin-layer Chromatography
Column: A stainless steel column 4 mm in inside diameter <2.03>. Spot 25 mL of the sample solution and 10 mL each of
and 25 cm in length, packed with octadecylsilanized silica gel the standard solutions (1) and (2) on a plate of silica gel with
for liquid chromatography (10 mm in particle diameter). fluorescent indicator for thin-layer chromatography. Pro-
Column temperature: A constant temperature of about ceed as directed in the Purity (2) under Chlordiazepoxide.
259 C.
Mobile phase: A mixture of methanol and 0.02 mol/L am-
monium dihydrogenphosphate TS (7:3).
Flow rate: Adjust so that the retention time of chlordia-
Conduct this procedure without exposure to light, using
zepoxide is about 5 minutes.
light-resistant vessels. To 1 tablet of Chlordiazepoxide
Tablets add 1 mL of water, shake to disintegrate the tablet,
then add 20 mL of methanol, shake, add methanol to make
mL of the standard solution under the above operating
exactly 25 mL, and filter through a membrane filter with a
conditions, chlordiazepoxide and the internal standard are
pore size not exceeding 0.5 mm. Discard the first 5 mL of the
filtrate, take exactly V mL of the subsequent filtrate equiva-
lent to about 2 mg of chlordiazepoxide (C16H14ClN3O), add
exactly 1 mL of the internal standard solution, then add
methanol to make 20 mL, and use this solution as the sample
of the peak area of chlordiazepoxide to that of the internal
standard is not more than 1.0z. Amount (mg) of chlordiazepoxide (C16H14ClN3O)
＝ MS × QT/QS × 5/V
Storage-Light-resistant. MS: Amount (mg) of Chlordiazepoxide RS taken
Internal standard solution—A solution of isobutyl salicylate
Chlordiazepoxide Tablets
クロルジアゼポキシド錠 lutions per minute according to the Paddle method, using
medium, the dissolution rate in 60 minutes of Chlordia-
Chlordiazepoxide Tablets contain not less than
zepoxide Tablets is not less than 70z.
Conduct this procedure without exposure to light, using
amount of chlordiazepoxide (C16H14ClN3O: 299.75).
light-resistant vessels. Start the test with 1 tablet of Chlordia-
Method of preparation Prepare as directed under Tablets, zepoxide Tablets, withdraw not less than 30 mL of the me-
with Chlordiazepoxide. dium at the specified minute after starting the test, and filter
Identification (1) Weigh a portion of powdered Chlordia-
0.8 mm. Discard the first 10 mL of the filtrate, pipet V mL of
zepoxide Tablets, equivalent to 0.01 g of Chlordiazepoxide,
the subsequent filtrate, add the dissolution medium to make
add 100 mL of 0.1 mol/L hydrochloric acid TS, shake, and
exactly V? mL so that each mL contains about 3.7 mg of
filter. To 5 mL of the filtrate add 0.1 mol/L hydrochloric
chlordiazepoxide (C16H14ClN3O), and use this solution as the
acid TS to make 100 mL, and determine the absorption spec-
trum of this solution as directed under Ultraviolet-visible
of Chlordiazepoxide RS, previously dried under reduced
pressure with phosphorus (V) oxide as a dessicant at 609C
nm and 248 nm and between 306 nm and 310 nm, and a
for 4 hours, dissolve in 5 mL of 0.1 mol/L hydrochloric acid
minimum between 288 nm and 292 nm.
TS, and add the dissolution medium to make exactly 200
(2) Weigh a portion of powdered Chlordiazepoxide
mL. Pipet 3 mL of this solution, add the dissolution medium
Tablets, equivalent to 0.01 g of Chlordiazepoxide, add 10
mL of diethyl ether, shake vigorously, and centrifuge.
Evaporate 5 mL of the supernatant liquid by warming on a
water bath to dryness. Determine the infrared absorption
under Ultraviolet-visible Spectrophotometry <2.24>.
spectrum of the residue as directed in the potassium bromide
disk method under Infrared Spectrophotometry <2.25>: it ex- Dissolution rate (z) with respect to the labeled amount
hibits absorption at the wave numbers of about 1625 cm－1, of chlordiazepoxide (C16H14ClN3O)
1465 cm－1, 1265 cm－1, 850 cm－1 and 765 cm－1. ＝ MS × AT/AS × V?/V × 1/C × 27
Purity Related substances—Conduct this procedure with- MS: Amount (mg) of Chlordiazepoxide RS taken
out exposure to light, using light-resistant vessels. To a por- C: Labeled amount (mg) of chlordiazepoxide
tion of powdered Chlordiazepoxide Tablets, equivalent to 50 (C16H14ClN3O) in 1 tablet
mg of Chlordiazepoxide, add exactly 5 mL of a mixture of
Assay Conduct this procedure without exposure to light,
methanol and ammonia TS (97:3), shake, centrifuge, and use
using light-resistant vessels. Weigh accurately a quantity of
the supernatant liquid as the sample solution. Separately,
Chlordiazepoxide Tablets, equivalent to about 0.1 g of
dissolve 50 mg of Chlordiazepoxide RS in a mixture of meth-
chlordiazepoxide (C16H14ClN3O), add 10 mL of water, and
anol and ammonia TS (97:3) to make exactly 50 mL, and use
shake well to disintegrate. Add 60 mL of methanol, shake
this solution as the standard solution (1). Dissolve 5.0 mg of
well, add methanol to make exactly 100 mL, and centrifuge.
682 Chlorhexidine Gluconate Solution / Official Monographs JP XVII
Pipet 10 mL of the supernatant liquid, add exactly 5 mL of Identification (1) To 0.05 mL of Chlorhexidine
the internal standard solution, add methanol to make exactly Gluconate Solution add 5 mL of methanol, 1 mL of bromine
100 mL, and use this solution as the sample solution. Sepa- TS and 1 mL of 8 mol/L sodium hydroxide TS: a deep red
rately, weigh accurately about 10 mg of Chlordiazepoxide color is produced.
RS, previously dried in a desiccator (in vacuum, phosphorus (2) To 0.5 mL of Chlorhexidine Gluconate Solution add
(V) oxide, 609C) for 4 hours, dissolve in 1 mL of water and a 10 mL of water and 0.5 mL of copper (II) sulfate TS: a white
suitable amount of methanol, add exactly 5 mL of the inter- precipitate is formed. Heat to boiling: the precipitate
nal standard solution, add methanol to make 100 mL, and changes to light purple.
use this solution as the standard solution. Perform the test (3) To 10 mL of Chlorhexidine Gluconate Solution add 5
with 10 mL each of the sample solution and standard solution mL of water, cool on ice, and add 5 mL of sodium hydrox-
as directed under Liquid Chromatography <2.01> according ide TS dropwise with stirring: a white precipitate is formed.
to the following conditions, and calculate the ratios, QT and Collect the precipitate by filtration, wash with water,
QS, of the peak area of chlordiazepoxide to that of the inter- recrystallize from diluted ethanol (95) (7 in 10), and dry at
nal standard. 1059C for 30 minutes: the crystals thus obtained melt <2.60>
between 1309 C and 1349 C.
Amount (mg) of chlordiazepoxide (C16H14ClN3O)
(4) Neutralize the filtrate obtained in (3) with 5 mol/L
＝ MS × QT/QS × 10
hydrochloric acid TS. To 5 mL of this solution add 0.65 mL
MS: Amount (mg) of Chlordiazepoxide RS taken of acetic acid (100) and 1 mL of freshly distilled phenyl-
hydrazine, and heat on a water bath for 30 minutes. After
Internal standard solution—A solution of isobutyl salicylate
cooling, scratch the inner wall of the vessel with a glass rod
to induce crystallization. Collect the crystals, dissolve in 10
mL of hot water, add a small amount of activated charcoal,
and filter. Cool the filtrate, scratch the inner side of the ves-
length: 254 nm).
sel, collect the formed crystals, and dry: the crystals thus ob-
tained melt <2.60> at about 1959C (with decomposition).
for liquid chromatography (10 mm in particle diameter). pH <2.54> To 5.0 mL of Chlorhexidine Gluconate Solution
Column temperature: A constant temperature of about add water to make 100 mL: the pH of the solution is between
259 C. 5.5 and 7.0.
Mobile phase: A mixture of methanol and 0.02 mol/L am-
Purity 4-Chloroaniline—To 2.0 mL of Chlorhexidine
monium dihydrogenphosphate TS (7:3).
Gluconate Solution add water to make exactly 100 mL. Pipet
Flow rate: Adjust so that the retention time of chlordia-
5 mL of the solution, and add 20 mL of water and 5 mL of 1
zepoxide is about 5 minutes.
mol/L hydrochloric acid TS. Add 0.3 mL of sodium nitrite
TS, shake, and allow to stand for 2 minutes. Add 4 mL of
ammonium amidosulfate TS, and then allow to stand for 1
minute. Add 5 mL of N-(1-naphthyl)-N?-diethylethylenedia-
ditions, chlordiazepoxide and the internal standard are
mine oxalate-acetone TS, allow to stand for 10 minutes, add
1 mL of ethanol (95), and then add water to make 50 mL:
the color of the solution is not more intense than the follow-
ing control solution.
Control solution: Dissolve 20 mg of 4-chloroaniline in 10
mL of 1 mol/L hydrochloric acid TS, and add water to make
of the peak area of chlordiazepoxide to that of the internal
exactly 100 mL. Pipet 5 mL of the solution, and add water to
make exactly 100 mL. Pipet 5 mL of the solution, add 20 mL
Containers and storage Containers—Tight containers. of water and 5 mL of 1 mol/L hydrochloric acid TS, and
proceed as directed for the preparation of the sample solu-
tion.
Chlorhexidine Gluconate Solution Residue on ignition <2.44> Not more than 0.1z (2 g, after
evaporation).
クロルヘキシジングルコン酸塩液
Assay Pipet 2 mL of Chlorhexidine Gluconate Solution,
evaporate to dryness on a water bath, dissolve the residue in
Chlorhexidine Gluconate Solution is a solution of
60 mL of acetic acid for nonaqueous titration, and titrate
digluconate of chlorhexidine. <2.50> with 0.1 mol/L perchloric acid VS (potentiometric
It contains not less than 19.0 w/vz and not
more than 21.0 w/vz of chlorhexidine gluconate
(C22H30Cl2N10.2C6H12O7: 897.76).
Description Chlorhexidine Gluconate Solution is a clear,
＝ 22.44 mg of C22H30Cl2N10.2C6H12O7
colorless or pale yellow liquid. It is odorless, and has a bitter
taste. Containers and storage Containers—Tight containers.
It is miscible with water and with acetic acid (100). 1 mL Storage—Light-resistant.
of Chlorhexidine Gluconate Solution is miscible with not
more than 5 mL of ethanol (99.5) and with not more than 3
mL of acetone. By further addition of each of these solvents,
a white turbidity is formed.
20: 1.06 – 1.07
JP XVII Official Monographs / Chlorinated Lime 683
of formic acid, 15 mL of 1 mol/L hydrochloric acid TS and

Chlorhexidine Hydrochloride 20 mL of water, and proceed in the same manner.
クロルヘキシジン塩酸塩
2 hours).
Assay Weigh accurately about 0.2 g of Chlorhexidine Hy-
drochloride, previously dried, dissolve in 2.0 mL of formic
acid, add 60 mL of acetic anhydride, and titrate <2.50> with
C22H30Cl2N10.2HCl: 578.37 form a blank determination, and make any necessary correc-
1,1?-Hexamethylenebis[5-(4-chlorophenyl)biguanide] tion.
dihydrochloride
[3697-42-5]
＝ 14.46 mg of C22H30Cl2N10.2HCl
Chlorhexidine Hydrochloride, when dried, contains Containers and storage Containers—Tight containers.
not less than 98.0z of chlorhexidine hydrochloride Storage—Light-resistant.
(C22H30Cl2N10.2HCl).
Description Chlorhexidine Hydrochloride occurs as a white
crystalline powder. It is odorless, and has a bitter taste. Chlorinated Lime
It is soluble in formic acid, slightly soluble in methanol
サラシ粉
and in warm methanol, and practically insoluble in water, in
It is gradually colored by light. Chlorinated Lime contains not less than 30.0z of
available chlorine (Cl: 35.45).
Identification (1) Dissolve 0.01 g of Chlorhexidine Hy-
drochloride in 5 mL of methanol by warming, and add 1 mL Description Chlorinated Lime occurs as a white powder. It
of bromine TS and 1 mL of 8 mol/L sodium hydroxide TS: a has a chlorine-like odor.
deep red color is produced. It dissolves partially in water. The solution changes red
(2) Dissolve 0.3 g of Chlorhexidine Hydrochloride in 10 litmus paper to blue, then gradually decolorizes.
mL of 6 mol/L hydrochloric acid TS, cool in ice, and add 10
Identification (1) To Chlorinated Lime add dilute hydro-
mL of 8 mol/L sodium hydroxide TS dropwise with stirring:
chloric acid: a gas, which has the odor of chlorine, evolves,
a white precipitate is produced. Collect the precipitate, wash
and the gas changes moistened starch-potassium iodide
with water, recrystallize from diluted ethanol (95) (7 in 10),
paper to blue.
and dry at 1059C for 30 minutes: the crystals so obtained
(2) Shake 1 g of Chlorinated Lime with 10 mL of water,
melt <2.60> between 1309 C and 1349C.
and filter: the filtrate responds to the Qualitative Tests
(3) Dissolve 0.1 g of Chlorhexidine Hydrochloride in 50
<1.09> (2) and (3) for calcium salt.
mL of dilute nitric acid: the solution responds to the Qualita-
tive Tests <1.09> for chloride. Assay Weigh accurately about 5 g of Chlorinated Lime,
transfer to a mortar, and triturate thoroughly with 50 mL of
water. Transfer to a 500-mL volumetric flask with the aid of
Chlorhexidine Hydrochloride according to Method 2, and
water, and add water to make 500 mL. Mix well, imme-
diately take exactly 50 mL of the mixture in an iodine flask,
add 10 mL of potassium iodide TS and 10 mL of dilute hy-
(2) Arsenic <1.11>—To 1.0 g of Chlorhexidine Hydro-
drochloric acid, and titrate <2.50> the liberated iodine with
chloride in a crucible add 10 mL of a solution of magnesium
0.1 mol/L sodium thiosulfate VS (indicator: 3 mL of starch
nitrate hexahydrate in ethanol (95) (1 in 10), fire the ethanol
TS). Perform a blank determination, and make any neces-
(95) to burn, and heat gradually to incinerate. If a car-
sary correction.
bonized substance remains, moisten with a small amount of
nitric acid, and ignite to incinerate. Cool, add 10 mL of Each mL of 0.1 mol/L sodium thiosulfate VS
dilute hydrochloric acid to the residue, dissolve by warming ＝ 3.545 mg of Cl
on a water bath, use this solution as the test solution, and
Storage—Light-resistant, and in a cold place.
(3) p-Chloroaniline—Dissolve 0.10 g of Chlorhexidine
Hydrochloride in 2 mL of formic acid, and add 15 mL of 1
mol/L hydrochloric acid TS and 20 mL of water immedi-
ately. Add 0.3 mL of sodium nitrite TS, shake, and allow to
stand for 2 minutes. Add 4 mL of ammonium amidosulfate
TS, and then allow to stand for 1 minute. Add 5 mL of N-
(1-naphthyl)-N?-diethylethylenediamine oxalate-acetone TS,
allow to stand for 10 minutes, and add 1 mL of ethanol (95)
and water to make 50 mL: the solution has no more color
than the following control solution.
Control solution: Dissolve 20 mg of 4-chloroaniline in 10
mL of 1 mol/L hydrochloric acid TS, and add water to make
exactly 100 mL. Pipet 5 mL of the solution, and add water to
make exactly 100 mL. To 2.0 mL of the solution add 2 mL
684 Chlormadinone Acetate / Official Monographs JP XVII
none acetate from the standard solution.
Chlormadinone Acetate Operating conditions—
クロルマジノン酢酸エステル length: 236 nm).
309C.
Mobile phase: A mixture of acetonitrile and water (13:7).
Flow rate: Adjust so that the retention time of chlormadi-
none acetate is about 10 minutes.
C23H29ClO4: 404.93 Time span of measurement: About 1.5 times as long as the
6-Chloro-3,20-dioxopregna-4,6-dien-17-yl acetate retention time of chlormadinone acetate, beginning after the
[302-22-7] solvent peak.
Chlormadinone Acetate, when dried, contains Test for required detectability: To exactly 5 mL of the
not less than 98.0z of chlormadinone acetate standard solution add acetonitorile to make exactly 50 mL.
(C23H29ClO4). Confirm that the peak area of chlormadinone acetate ob-
Description Chlormadinone Acetate occurs as white to
of that of chlormadinone acetate obtained from 10 mL of the
light yellow, crystals or crystalline powder. It is odorless.
standard solution.
It is freely soluble in chloroform, soluble in acetonitrile,
System performance: Dissolve 8 mg of Chlormadinone
slightly soluble in ethanol (95) and in diethyl ether, and prac-
Acetate and 2 mg of butyl parahydroxybenzoate in 100 mL
tically insoluble in water.
of acetonitrile. When the procedure is run with 10 mL of this
Identification (1) Dissolve 2 mg of Chlormadinone solution under the above operating conditions, butyl parahy-
Acetate in 1 mL of ethanol (95), and add 1 mL of 1,3- droxybenzoate and chlormadinone acetate are eluted in this
dinitrobenzene TS and 1 mL of a solution of potassium hy- order with the resolution between these peaks being not less
droxide (1 in 5): a red-purple color develops. than 8.
(2) To 0.05 g of Chlormadinone Acetate add 2 mL of po- System repeatability: When the test is repeated 6 times
tassium hydroxide-ethanol TS, and boil on a water bath for with 10 mL of the standard solution under the above operat-
5 minutes. After cooling, add 2 mL of diluted sulfuric acid ing conditions, the relative standard deviation of the peak
(2 in 7), and boil gently for 1 minute: the odor of ethyl ace- area of chlormadinone acetate is not more than 1.0z.
tate is perceptible.
Chlormadinone Acetate, previously dried, as directed in the
potassium bromide disk method under Infrared Spectropho- Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Assay Weigh accurately about 20 mg each of Chlormadi-
ence Spectrum or the spectrum of previously dried Chlor-
none Acetate and Chlormadinone Acetate RS, previously
madinone Acetate RS: both spectra exhibit similar intensities
dried, and dissolve in ethanol (95) to make exactly 100 mL.
Pipet 5 mL each of these solutions, to each add ethanol (95)
(4) Perform the test with Chlormadinone Acetate as di-
to make exactly 100 mL, and use these solutions as the sam-
ple solution and the standard solution, respectively. Perform
appears.
the test with these solutions as directed under Ultraviolet-
Optical rotation <2.49> [a]20 D : －10.0 – －14.09(after dry- visible Spectrophotometry <2.24>, and determine the absor-
ing, 0.2 g, acetonitrile, 10 mL, 100 mm). bances, AT and AS, at 285 nm.
Melting point <2.60> 211 – 2159C Amount (mg) of chlormadinone acetate (C23H29ClO4)
＝ M S × A T / AS
Chlormadinone Acetate according to Method 2, and per- MS: Amount (mg) of Chlormadinone Acetate RS taken
of Chlormadinone Acetate according to Method 3, and per-
(3) Related substances—Dissolve 20 mg of Chlormadi-
none Acetate in 10 mL of acetonitrile, and use this solution
add acetonitrile to make exactly 100 mL, and use this solu-
the automatic integration method: the total area of peaks
other than the peak of chlormadinone acetate from the sam-
ple solution is not larger than the peak area of chlormadi-
JP XVII Official Monographs / Chlorphenesin Carbamate 685
Chlorobutanol Chlorphenesin Carbamate

クロロブタノールクロルフェネシンカルバミン酸エステル
C4H7Cl3O: 177.46
C10H12ClNO4: 245.66
1,1,1-Trichloro-2-methylpropan-2-ol
(2RS )-3-(4-Chlorophenoxy)-2-hydroxypropyl carbamate
[57-15-8]
[886-74-8]
Chlorobutanol contains not less than 98.0z of chlo-
Chlorphenesin Carbamate, when dried, contains
robutanol (C4H7Cl3O), calculated on the anhydrous
basis.
chlorphenesin carbamate (C10H12ClNO4).
Description Chlorobutanol occurs as colorless or white
Description Chlorphenesin Carbamate occurs as white,
crystals. It has a camphoraceous odor.
crystals or a crystalline powder.
It is very soluble in methanol, in ethanol (95) and in
It is freely soluble in methanol, in ethanol (95) and in pyri-
diethyl ether, and slightly soluble in water.
dine, and slightly soluble in water.
It slowly volatilizes in air.
A solution of Chlorphenesin Carbamate in ethanol (95)
Melting point: not lower than about 769C.
(1 in 20) shows no optical rotation.
Identification (1) To 5 mL of a solution of Chlo-
robutanol (1 in 200) add 1 mL of sodium hydroxide TS, then
solution of Chlorphenesin Carbamate in ethanol (95) (3 in
slowly add 3 mL of iodine TS: a yellow precipitate is pro-
duced and the odor of iodoform is perceptible.
(2) To 0.1 g of Chlorobutanol add 5 mL of sodium hy-
droxide TS, shake well the mixture, add 3 to 4 drops of ani-
line, and warm gently: the disagreeable odor of phenyl isoc-
yanide (poisonous) is perceptible.
Chlorphenesin Carbamate, as directed in the potassium bro-
Purity (1) Acidity—Shake thoroughly 0.10 g of the pow- mide disk method under Infrared Spectrophotometry <2.25>,
der of Chlorobutanol with 5 mL of water: the solution is and compare the spectrum with the Reference Spectrum:
neutral. both spectra exhibit similar intensities of absorption at the
(2) Chloride <1.03>—Dissolve 0.5 g of Chlorobutanol in same wave numbers.
25 mL of dilute ethanol, and add 6 mL of dilute nitric acid (3) Perform the test with Chlorphenesin Carbamate as
and water to make 50 mL. Perform the test using this solu- directed under Flame Coloration Test <1.04> (2): a green
tion as the test solution. Prepare the control solution with color appears.
1.0 mL of 0.01 mol/L hydrochloric acid VS by adding 25
mL of dilute ethanol, 6 mL of dilute nitric acid and water to
make 50 mL (not more than 0.071z). Purity (1) Heavy metals <1.07>—Dissolve 2.0 g of Chlor-
phenesin Carbamate in 20 mL of ethanol (95), and add 2 mL
of dilute acetic acid and water to make 50 mL. Perform the
Residue on ignition <2.44> Not more than 0.1z (1 g). trol solution as follows: to 2.0 mL of Standard Lead Solu-
tion add 20 mL of ethanol (95), 2 mL of dilute acetic acid
Assay Transfer about 0.1 g of Chlorobutanol, accurately
and water to make 50 mL (not more than 10 ppm).
weighed, to a 200-mL conical flask, and dissolve in 10 mL of
ethanol (95). Add 10 mL of sodium hydroxide TS, boil
of Chlorphenesin Carbamate according to Method 3, and
under a reflux condenser for 10 minutes, cool, add 40 mL of
dilute nitric acid and exactly 25 mL of 0.1 mol/L silver ni-
(3) Chlorphenesin-2-carbamate—Dissolve 0.10 g of
trate VS, and shake well. Add 3 mL of nitrobenzene, and
Chlorphenesin Carbamate in 20 mL of a mixture of hexane
shake vigorously until the precipitate is coagulated. Titrate
for liquid chromatography and 2-propanol (7:3), and use
<2.50> the excess silver nitrate with 0.1 mol/L ammonium
this solution as the sample solution. Perform the test with 10
thiocyanate VS (indicator: 2 mL of ammonium iron (III) sul-
mL of the sample solution as directed under Liquid Chroma-
fate TS). Perform a blank determination.
tography <2.01> according to the following conditions. De-
Each mL of 0.1 mol/L silver nitrate VS termine the peak area, Aa, of chlorphenesin carbamate and
＝ 5.915 mg of C4H7Cl3O the peak area, Ab, of chlorphenesin-2-carbamate by the au-
tomatic integration method: the ratio, Ab/(Aa ＋ Ab), is not
more than 0.007.
length: 280 nm).
and 30 cm in length, packed with silica gel for liquid chro-
matography (5 mm in particle diameter).
686 Chlorphenesin Carbamate Tablets / Official Monographs JP XVII
409 C. Each mL of 0.1 mol/L potassium hydroxide-ethanol TS
Mobile phase: A mixture of hexane for liquid chromatog- ＝ 24.57 mg of C10H12ClNO4
raphy, 2-propanol and acetic acid (100) (700:300:1).
Flow rate: Adjust so that the retention time of chlorphene-
sin carbamate is about 9 minutes.
Test for required detectability: To 1 mL of the sample so- Chlorphenesin Carbamate Tablets
lution, add a mixture of hexane for liquid chromatography
クロルフェネシンカルバミン酸エステル錠
and 2-propanol (7:3) to make 100 mL, and use this solution
as the solution for system suitability test. To exactly 5 mL of
the solution for system suitability test add the mixture of Chlorphenesin Carbamate Tablets contain not less
hexane for liquid chromatography and 2-propanol (7:3) to than 93.0z and not more than 107.0z of the labeled
make exactly 10 mL. Confirm that the peak area of chlor- amount of chlorphenesin carbamate (C10H12ClNO4:
phenesin carbamate obtained from 10 mL of this solution is 245.66).
equivalent to 40 to 60z of that of chlorphenesin carbamate
obtained from 10 mL of the solution for system suitability
with Chlorphenesin Carbamate.
test.
System performance: Dissolve 0.1 g of Chlorphenesin Identification To a quantity of powdered Chlorphenesin
Carbamate in 50 mL of methanol. To 25 mL of this solution Carbamate Tablets, equivalent to 0.15 g of Chlorphenesin
add 25 mL of dilute sodium hydroxide TS, and warm at Carbamate, add 60 mL of ethanol (95), treat with ultrasonic
609 C for 20 minutes. To 20 mL of this solution add 5 mL of waves, and add ethanol (95) to make 100 mL. Centrifuge 20
1 mol/L hydrochloric acid TS, shake well with 20 mL of mL of this solution, add ethanol (95) to 1 mL of the superna-
ethyl acetate, and allow to stand to separate the upper layer. tant liquid to make 100 mL, and determine the absorption
When the procedure is run with 10 mL of this layer under the spectrum of this solution as directed under Ultraviolet-
above operating conditions, chlorphenesin, chlorphenesin visible Spectrophotometry <2.24>: it exhibits maxima be-
carbamate and chlorphenesin-2-carbamate are eluted in this tween 226 nm and 230 nm, between 279 nm and 283 nm, and
order, with the relative retention times of chlorphenesin and between 286 nm and 290 nm.
chlorphenesin-2-carbamate to chlorphenesin carbamate
being about 0.7 and about 1.2, respectively, and with the
resolution between the peaks of chlorphenesin and chlor-
phenesin carbamate being not less than 2.0.
To 1 tablet of Chlorphenesin Carbamate Tablets add 10
mL of water to disintegrate the tablet, add 70 mL of a mix-
ture of water and methanol (1:1), treat with ultrasonic waves
for 15 minutes with occasional stirring, then add the mixture
tion of the peak areas of chlorphenesin carbamate is not
of water and methanol (1:1) to make exactly 100 mL. Centri-
more than 2.0z.
fuge this solution, pipet V mL of the supernatant liquid
(4) Related substances—Dissolve 0.10 g of Chlorphene-
equivalent to about 2.5 mg of chlorphenesin carbamate
sin Carbamate in 10 mL of ethanol (95), and use this solu-
(C10H12ClNO4), add the mixture of water and methanol (1:1)
to make exactly 25 mL, and use this solution as the sample
tion, add ethanol (95) to make exactly 20 mL. Pipet 2 mL of
solution. Separately, weigh accurately about 50 mg of chlor-
this solution, add ethanol (95) to make exactly 20 mL, and
phenesin carbamate for assay, previously dried in a desicca-
tor (in vacuum, silica gel) for 4 hours, and dissolve in the
with these solutions as directed under Thin-layer Chroma-
mixture of water and methanol (1:1) to make exactly 50 mL.
tography <2.03>. Spot 50 mL each of the sample solution and
Pipet 2 mL of this solution, add the mixture of water and
standard solution on a plate of silica gel for thin-layer chro-
methanol (1:1) to make exactly 20 mL, and use this solution
matography. Develop the plate with a mixture of ethyl ace-
as the standard solution. Determine the absorbances at 280
tate, methanol and ammonia solution (28) (17:2:1) to a dis-
nm, AT and AS, of the sample solution and standard solution
tance of about 10 cm, and air-dry the plate. Allow the plate
to stand in iodine vapor for 20 minutes: the spot other than
<2.24>.
the principal spot from the sample solution is not more than
one, and it is not more intense than the spot from the stand- Amount (mg) of chlorphenesin carbamate
ard solution. (C10H12ClNO4)
＝ MS × AT/AS × 1/V × 5
um, silica gel, 4 hours). MS: Amount (mg) of chlorphenesin carbamate for assay
taken
Assay Weigh accurately about 0.5 g of Chlorphenesin Car-
bamate, previously dried, dissolve in 20 mL of pyridine, add
exactly 50 mL of 0.1 mol/L potassium hydroxide-ethanol
in 15 minutes of Chlorphenesin Carbamate Tablets is not
TS, and warm at 709C for 40 minutes. After cooling, add
less than 85z.
100 mL of ethanol (95), and titrate <2.50> the excess potas-
Start the test with 1 tablet of Chlorphenesin Carbamate
sium hydroxide with 0.1 mol/L hydrochloric acid VS until
the color of the solution changes from blue through blue-
green to yellow (indicator: 1 mL of thymol blue TS). Per-
form a blank determination in the same manner.
JP XVII Official Monographs / Chlorpheniramine Maleate 687
mL contains about 0.14 mg of chlorphenesin carbamate ternal standard is not more than 1.5z.
(C10H12ClNO4), and use this solution as the sample solution.
Separately, weigh accurately about 28 mg of chlorphenesin
ers.
carbamate for assay, previously dried in a desiccator (in
vacuum, silica gel) for 4 hours, dissolve in 1 mL of metha-
nol, and add water to make exactly 50 mL. Pipet 5 mL of
this solution, add water to make exactly 20 mL, and use this Chlorpheniramine Maleate
solution as the standard solution. Determine the absor-
クロルフェニラミンマレイン酸塩
bances, AT and AS, at 278 nm of the sample solution and
of chlorphenesin carbamate (C10H12ClNO4)
＝ MS × AT/AS × V?/V × 1/C × 450
MS: Amount (mg) of chlorphenesin carbamate for assay
C16H19ClN2.C4H4O4: 390.86
taken
(3RS )-3-(4-Chlorophenyl)-N, N-dimethyl-3-pyridin-
C: Labeled amount (mg) of chlorphenesin carbamate
2-ylpropylamine monomaleate
(C10H12ClNO4) in 1 tablet
[113-92-8]
Assay Weigh accurately the mass of not less than 20 Chlor-
phenesin Carbamate Tablets, and powder them in an agate Chlorpheniramine Maleate, when dried, contains
mortar. Weigh accurately a portion of the powder, not less than 98.0z and not more than 101.0z of dl-
equivalent to about 0.25 g of chlorphenesin carbamate chlorpheniramine maleate (C16H19ClN2.C4H4O4).
(C10H12ClNO4), add 30 mL of ethyl acetate, disperse using
Description Chlorpheniramine Maleate occurs as white,
ultrasonic waves, then add ethyl acetate to make exactly 50
fine crystals.
mL. Centrifuge 20 mL of this solution, pipet 2 mL of the su-
pernatant liquid, add exactly 2 mL of the internal standard
water and in methanol, and soluble in ethanol (99.5).
solution, add ethyl acetate to make 20 mL, and use this
A solution of Chlorpheniramine Maleate (1 in 20) shows
about 0.1 g of chlorphenesin carbamate for assay, previously
dried in a desiccator (in vacuum, silica gel) for 4 hours, and
dissolve in ethyl acetate to make exactly 50 mL. Pipet 5 mL Identification (1) Determine the absorption spectrum of a
of this solution, add exactly 2 mL of the internal standard solution of Chlorpheniramine Maleate in 0.1 mol/L hydro-
solution, then add ethyl acetate to make 20 mL, and use this chloric acid TS (3 in 100,000) as directed under Ultraviolet-
solution as the standard solution. Perform the test with 10 visible Spectrophotometry <2.24>, and compare the spectrum
mL each of the sample solution and standard solution as di- with the Reference Spectrum or the spectrum of a solution of
rected under Liquid Chromatography <2.01> according to Chlorpheniramine Maleate RS prepared in the same manner
the following conditions, and calculate the ratios, QT and as the sample solution: both spectra exhibit similar intensi-
QS, of the peak area of chlorphenesin carbamate to that of ties of absorption at the same wavelengths.
the internal standard. (2) Determine the infrared absorption spectrum of
Chlorpheniramine Maleate, previously dried, as directed in
Amount (mg) of chlorphenesin carbamate (C10H12ClNO4)
the paste method under Infrared Spectrophotometry <2.25>,
＝ MS × QT/QS × 5/2
MS: Amount (mg) of chlorphenesin carbamate for assay the spectrum of previously dried Chlorpheniramine Maleate
taken RS: both spectra exhibit similar intensities of absorption at
Internal standard solution—A solution of ethenzamide in
(3) Dissolve 0.10 g of Chlorpheniramine Maleate in 5 mL
ethyl acetate (1 in 400).
Separately, dissolve 56 mg of maleic acid in 10 mL of metha-
nol, and use this solution as the standard solution. Perform
length: 280 nm).
and 30 cm in length, packed with silica gel for liquid chro-
tion and standard solution on a plate of silica gel for thin-
matography (5 mm in particle diameter).
diethyl ether, methanol, acetic acid (100) and water
409 C.
Mobile phase: A mixture of hexane for liquid chromatog-
raphy, 2-propanol and acetic acid (100) (700:300:1).
nm): a spot among two of the spots obtained with the sample
Flow rate: Adjust so that the retention time of chlorphene-
solution shows the same intense and R f value with the spot
sin carbamate is about 9 minutes.
obtained with the standard solution.
System performance: Proceed as directed in the system pH <2.54> Dissolve 1.0 g of Chlorpheniramine Maleate in
suitability in the Purity (3) under Chlorphenesin Carbamate. 100 mL of freshly boiled and cooled water: the pH of this so-
System repeatability: When the test is repeated 6 times lution is between 4.0 and 5.5.
the peak area of chlorphenesin carbamate to that of the in- Purity (1) Clarity and color of solution—Dissolve 1.0 g
688 Chlorpheniramine Maleate Injection / Official Monographs JP XVII
of Chlorpheniramine Maleate in 50 mL of water: the solu- Each mL of 0.1 mol/L perchloric acid VS
tion is clear and colorless. ＝ 19.54 mg of C16H19ClN2.C4H4O4
(2) Heavy metals <1.07>—Proceed with 1.0 g of Chlor-
pheniramine Maleate according to Method 4, and perform
(3) Related substances—Dissolve 0.10 g of Chlor-
pheniramine Maleate in 100 mL of the mobile phase, and use Chlorpheniramine Maleate Injection
クロルフェニラミンマレイン酸塩注射液
ple solution, and add the mobile phase to make exactly 100
mL. Pipet 2 mL of this solution, add the mobile phase to
make exactly 20 mL, and use this solution as the standard Chlorpheniramine Maleate Injection is an aqueous
solution. Perform the test with exactly 20 mL each of the injection.
sample solution and standard solution as directed under Liq- It contains not less than 95.0z and not more than
uid Chromatography <2.01> according to the following con- 105.0z of the labeled amount of dl-chlorpheniramine
ditions, and determine each peak area by the automatic in- maleate (C16H19ClN2.C4H4O4: 390.86).
tegration method: the area of the peak other than maleic
acid and chlorpheniramine obtained with the sample solu-
tions, with Chlorpheniramine Maleate.
tion is not larger than 2/3 times the peak area of chlor-
pheniramine obtained with the standard solution, and the Description Chlorpheniramine Maleate Injection is a clear,
total area of the peaks other than maleic acid and chlor- colorless liquid.
pheniramine with the sample solution is not larger than the pH: 4.5 – 7.0
peak area of chlorpheniramine with the standard solution.
Identification Take a volume of Chlorpheniramine Male-
ate Injection, equivalent to 25 mg of Chlorpheniramine
Maleate, add 5 mL of dilute sodium hydroxide TS, and ex-
length: 225 nm).
tract with 20 mL of hexane. Wash the hexane layer with 10
mL of water, shake with 0.5 g of anhydrous sodium sulfate
for several minutes, and filter. Evaporate the filtrate in a
water bath at 509 C under a reduced pressure, and determine
the infrared absorption spectrum of the residue as directed in
259 C.
the liquid film method under Infrared Spectrophotometry
<2.25>: it exhibits absorption at the wave numbers of about
phosphate and 1 mL of phosphoric acid in water to make
2940 cm－1, 2810 cm－1, 2770 cm－1, 1589 cm－1, 1491 cm－1,
1470 cm－1, 1434 cm－1, 1091 cm－1 and 1015 cm－1.
nitrile.
Flow rate: Adjust so that the retention time of chlor- Bacterial endotoxins <4.01> Less than 8.8 EU/mg.
pheniramine is about 11 minutes.
retention time of chlorpheniramine, beginning after the sol- Foreign insoluble matter <6.06> Perform the test according
vent peak. to Method 1: it meets the requirement.
Test for required detectability: To exactly 2.5 mL of the
ment.
mL. Confirm that the peak area of chlorpheniramine ob- Sterility <4.06> Perform the test according to the Mem-
tained with 20 mL of this solution is equivalent to 7 to 13z brane filtration method: it meets the requirement.
of that obtained with 20 mL of the standard solution.
Assay Transfer an exactly measured volume of Chlor-
pheniramine Maleate Injection, equivalent to about 3 mg of
chlorpheniramine maleate (C16H19ClN2.C4H4O4), to a
100-mL separator, add 20 mL of water and 2 mL of sodium
factor of the peak of chlorpheniramine are not less than 4000
hydroxide TS, and extract with two 50-mL portions of
diethyl ether. Combine the diethyl ether extracts, wash with
20 mL of water, and then extract with 20-mL, 20-mL and
5-mL portions of 0.25 mol/L sulfuric acid TS successively.
Combine all acid extracts, and add 0.25 mol/L sulfuric acid
area of chlorpheniramine is not more than 4.0z.
TS to make exactly 50 mL. Pipet 10 mL of this solution, add
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 0.25 mol/L sulfuric acid TS to make exactly 25 mL, and use
3 hours). this solution as the sample solution. Separately, weigh accu-
rately about 30 mg of Chlorpheniramine Maleate RS, previ-
ously dried at 1059C for 3 hours, and dissolve in water to
Assay Dissolve about 0.4 g of Chlorpheniramine Maleate, make exactly 200 mL. Pipet 20 mL of this solution, transfer
previously dried and accurately weighed, in 20 mL of acetic to a 100-mL separator, add 2 mL of sodium hydroxide TS,
acid (100). Titrate <2.50> with 0.1 mol/L perchloric acid VS and extract with two 50-mL portions of diethyl ether. Pro-
until the color of the solution changes from purple through ceed in the same manner as for the preparation of the sample
blue-green to green (indicator: 2 drops of crystal violet TS). solution, and use the solution so obtained as the standard so-
Perform a blank determination, and make any necessary lution. Determine the absorbances AT and AS of the sample
correction. solution and standard solution at a wavelength of the maxi-
JP XVII Official Monographs / Chlorpheniramine Maleate Powder 689
mum absorbance at about 265 nm as directed under Ultravi- taken

olet-visible Spectrophotometry <2.24>. C: Labeled amount (mg) of chlorpheniramine maleate
(C16H19ClN2.C4H4O4) in 1 g
Amount (mg) of chlorpheniramine maleate
(C16H19ClN2.C4H4O4) Operating conditions—
＝ MS × AT/AS × 1/10 Proceed as directed in the operating conditions in the
Assay.
MS: Amount (mg) of Chlorpheniramine Maleate RS taken
Containers and storage Containers—Hermetic containers. System performance: When the procedure is run with 50
Storage—Light-resistant. mL of the standard solution under the above operating con-
Chlorpheniramine Maleate Powder and not more than 2.5, respectively.
クロルフェニラミンマレイン酸塩散 with 50 mL of the standard solution under the above operat-
Chlorpheniramine Maleate Powder contains not less
than 93.0z and not more than 107.0z of the labeled Assay Weigh accurately an amount of Chlorpheniramine
amount of dl-chlorpheniramine maleate Maleate Powder, equivalent to about 4 mg of chlorphenira-
(C16H19ClN2.C4H4O4: 390.86). mine maleate (C16H19ClN2.C4H4O4), add 70 mL of the inter-
nal standard solution, shake for 15 minutes, then add the in-
Method of preparation Prepare as directed under Granules
ternal standard solution to make exactly 100 mL, and use
or Powders, with Chlorpheniramine Maleate.
Identification Weigh a portion of Chlorpheniramine rately about 20 mg of Chlorpheniramine Maleate RS, previ-
Maleate Powder, equivalent to 50 mg of Chlorpheniramine ously dried at 1059C for 3 hours, and add the internal stand-
Maleate, shake with 40 mL of 0.1 mol/L hydrochloric acid ard to make exactly 100 mL. Pipet 20 mL of this solution,
TS, and filter. Transfer the filtrate to a separator, and wash add the internal standard solution to make exactly 100 mL,
with 40 mL of hexane. Add 10 mL of sodium hydroxide TS, and use this solution as the standard solution. Perform the
and extract with 20 mL of hexane. Wash the hexane layer test with 30 mL each of the sample solution and standard so-
with 5 mL of water. Centrifuge, if necessary, shake the lution as directed under Liquid Chromatography <2.01> ac-
hexane extract with 0.5 g of anhydrous sodium sulfate for cording to the following conditions, and calculate the ratios,
several minutes, and filter. Evaporate the filtrate in a water QT and QS, of the peak area of chlorpheniramine to that of
bath at about 509C under reduced pressure, and determine the internal standard.
the infrared absorption spectrum of the residue as directed in
the liquid film method under Infrared Spectrophotometry
(C16H19ClN2.C4H4O4)
<2.25>: it exhibits absorption at the wave number of about
＝ MS × QT/QS × 1/5
2940 cm－1, 2810 cm－1, 2770 cm－1, 1589 cm－1, 1491 cm－1,
1470 cm－1, 1434 cm－1, 1091 cm－1 and 1015 cm－1. MS: Amount (mg) of Chlorpheniramine Maleate RS taken
Dissolution <6.10> When the test is performed at 50 revolu- Internal standard solution—To 7 mL of a solultion of
tions per minute according to the Paddle method, using 900 methyl parahydroxybenzoate in methanol (1 in 1000) add
mL of water as the dissolution medium, the dissolution rate water to make 1000 mL.
in 15 minutes of Chlorpheniramine Maleate Powder is not Operating conditions—
less than 85z. Detector: An ultraviolet absorption photometer (wave-
Start the test with an accurately weighed amount of Chlor- length: 265 nm).
pheniramine Maleate Powder, equivalent to about 4 mg of Column: A stainless steel column 4.6 mm in inside diame-
chlorpheniramine maleate (C16H19ClN2.C4H4O4), withdraw ter and 15 cm in length, packed with octadecylsilanized silica
not less than 20 mL of the medium at the specified minute gel for liquid chromatography (5 mm in particle diameter).
after starting the test, and filter through a membrane filter Column temperature: A constant temperature of about
with a pore size not exceeding 0.45 mm. Discard the first 10 409C.
mL of the filtrate, and use the subsequent filtrate as the sam- Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate
ple solution. Separately, weigh accurately about 22 mg of in 900 mL of water, add 10 mL of acetic acid (100) and water
Chlorpheniramine Maleate RS, previously dried at 1059 C to make 1000 mL. To 650 mL of this solution add 350 mL of
for 3 hours, and dissolve in water to make exactly 100 mL. acetonitrile.
Pipet 2 mL of this solution, add water to make exactly 100 Flow rate: Adjust so that the retention time of chlor-
mL, and use this solution as the standard solution. Perform pheniramine is about 8 minutes.
the test with exactly 50 mL each of the sample solution and System suitability—
standard solution as directed under Liquid Chromatography System performance: When the procedure is run with 30
<2.01> according to the following conditions, and determine mL of the standard solution under the above operating
the peak areas, AT and AS, of chlorpheniramine in each solu- conditions, the internal standard and chlorpheniramine are
tion. eluted in this order with the resolution between these peaks
of chlorpheniramine maleate (C16H19ClN2.C4H4O4)
＝ MS/MT × AT/AS × 1/C × 18
MS: Amount (mg) of Chlorpheniramine Maleate RS taken the peak area of chlorpheniramine to that of the internal
MT: Amount (g) of Chlorpheniramine Maleate Powder standard is not more than 1.0z.
690 Chlorpheniramine Maleate Tablets / Official Monographs JP XVII
Containers and storage Containers—Tight containers. membrane filter with a pore size not exceeding 0.45 mm. Dis-
Chlorpheniramine Maleate Tablets mL contains about 4.4 mg of chlorpheniramine maleate
(C16H19ClN2.C4H4O4), and use this solution as the sample
クロルフェニラミンマレイン酸塩錠 solution. Separately, weigh accurately about 22 mg of Chlor-
pheniramine Maleate RS, previously dried at 1059C for 3
hours, and dissolve in water to make exactly 100 mL. Pipet 2
Chlorpheniramine Maleate Tablets contain not
less than 93.0z and not more than 107.0z of the
labeled amount of dl-chlorpheniramine maleate
(C16H19ClN2.C4H4O4: 390.86).
solution as directed under Liquid Chromatography <2.01>,
Method of preparation Prepare as directed under Tablets, and determine the peak areas, AT and AS, of chlorphenira-
with Chlorpheniramine Maleate. mine in each solution.
Identification Weigh a portion of powdered Chlorphenira- Dissolution rate (z) with respect to the labeled amount
mine Maleate Tablets, equivalent to 50 mg of Chlorphenira- of chlorpheniramine maleate (C16H19ClN2.C4H4O4)
mine Maleate, shake with 40 mL of 0.1 mol/L hydrochloric ＝ MS × AT/AS × V?/V × 1/C × 18
acid TS, and filter. Transfer the filtrate to a separator, and
wash with 40 mL of hexane. Add 10 mL of sodium hydrox-
C: Labeled amount (mg) of chlorpheniramine maleate
ide TS, and extract with 20 mL of hexane. Wash the hexane
(C16H19ClN2.C4H4O4) in 1 tablet
layer with 5 mL of water. Centrifuge, if necessary, shake the
hexane extract with 0.5 g of anhydrous sodium sulfate for Operating conditions—
several minutes, and filter. Evaporate the filtrate in a water Detector: An ultraviolet absorption photometer (wave-
bath at about 509C under a reduced pressure, and determine length: 265 nm).
the infrared absorption spectrum of the residue as directed in Column: A stainless steel column 4.6 mm in inside diame-
the liquid film method under Infrared Spectrophotometry ter and 15 cm in length, packed with octadecylsilanized silica
<2.25>: it exhibits absorption at the wave numbers of about gel for liquid chromatography (5 mm in particle diameter).
2940 cm－1, 2810 cm－1, 2770 cm－1, 1589 cm－1, 1491 cm－1, Column temperature: A constant temperature of about
1470 cm－1, 1434 cm－1, 1091 cm－1 and 1015 cm－1. 409C.
Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate
in 900 mL of water, add 10 mL of acetic acid (100), and add
water to make 1000 mL. To 650 mL of this solution add 350
mL of acetonitrile.
To 1 tablet of Chlorpheniramine Maleate Tablets add 10
Flow rate: Adjust so that the retention time of chlor-
mL of water, shake to disintegrate the tablet, then add water
to make exactly V mL of a solution containing about 80 mg
of chlorpheniramine maleate (C16H19ClN2.C4H4O4) per mL,
exceeding 0.5 mm. Pipet 5 mL of the filtrate, add exactly 2.5
mL of the internal standard solution, add water to make 10
weigh accurately about 20 mg of Chlorpheniramine Maleate
RS, previously dried at 1059 C for 3 hours, and add water to
make exactly 100 mL. Pipet 20 mL of this solution, add ex-
actly 25 mL of the internal standard solution, add water to
Perform the test with 30 mL each of the sample solution and Assay Weigh accurately the mass of not less than 20 Chlor-
standard solution as directed under Liquid Chromatography pheniramine Maleate Tablets, and powder. Weigh accurately
<2.01> according to the conditions described in the Assay, a portion of the powder, equivalent to about 4 mg of chlor-
and calculate the ratios, QT and QS, of the peak area of pheniramine maleate (C16H19ClN2.C4H4O4), add 70 mL of
chlorpheniramine to that of the internal standard. the internal standard solution, shake for 15 minutes, then
add the internal standard solution to make exactly 100 mL,
filter through a membrane filter with a pore size not exceed-
(C16H19ClN2.C4H4O4)
ing 0.5 mm, and use this solution as the sample solution.
＝ MS × QT/QS × V/250
Separately, weigh accurately about 20 mg of Chlorphenira-
MS: Amount (mg) of Chlorpheniramine Maleate RS taken mine Maleate RS, previously dried at 1059C for 3 hours, and
add the internal standard to make exactly 100 mL. Pipet 20
Internal standard solution—To 7 mL of a solution of methyl
mL of this solution, add the internal standard solution to
parahydroxybenzoate (1 in 250) add water to make 1000 mL.
Dissolution <6.10> When the test is performed at 50 revolu- solution. Perform the test with 30 mL each of the sample so-
tions per minute according to the Paddle method, using 900 lution and standard solution as directed under Liquid Chro-
mL of water as the dissolution medium, the dissolution rate matography <2.01> according to the following conditions,
in 45 minutes of Chlorpheniramine Maleate Tablets is not and calculate the ratios, QT and QS, of the peak area of
less than 75z. chlorpheniramine to that of the internal standard.
Start the test with 1 tablet of Chlorpheniramine Maleate
JP XVII Official Monographs / d-Chlorpheniramine Maleate 691
Amount (mg) of chlorpheniramine maleate similar intensities of absorption at the same wavelengths.
(C16H19ClN2.C4H4O4) (2) Determine the infrared absorption spectrum of d-
＝ MS × QT/QS × 1/5 Chlorpheniramine Maleate, previously dried, as directed in
the paste method under Infrared Spectrophotometry <2.25>,
Internal standard solution—To 7 mL of a solution of methyl both spectra exhibit similar intensities of absorption at the
parahydroxybenzoate in methanol (1 in 1000) add water to same wave numbers.
make 1000 mL. (3) Dissolve 0.10 g of d-Chlorpheniramine Maleate in 5
Operating conditions— mL of methanol, and use this solution as the sample solu-
Detector: An ultraviolet absorption photometer (wave- tion. Separately, dissolve 56 mg of maleic acid in 10 mL of
length: 265 nm). methanol, and use this solution as the standard solution.
Column: A stainless steel column 4.6 mm in inside diame- Perform the test with these solutions as directed under Thin-
ter and 15 cm in length, packed with octadecylsilanized silica layer Chromatography <2.03>. Spot 5 mL each of the sample
gel for liquid chromatography (5 mm in particle diameter). solution and standard solution on a plate of silica gel for
Column temperature: A constant temperature of about thin-layer chromatography. Develop the plate with a mixture
409 C. of diethyl ether, methanol, acetic acid (100) and water
Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate (70:20:7:3) to a distance of about 12 cm, and air-dry the
in 900 mL of water, add 10 mL of acetic acid (100) and water plate. Examine under ultraviolet light (main wavelength: 254
to make 1000 mL. To 650 mL of this solution add 350 mL of nm): a spot among two of the spots obtained with the sample
acetonitrile. solution shows the same intense to the spot obtained with the
Flow rate: Adjust so that the retention time of chlor- standard solution, and its R f value is about 0.4.
D : ＋39.5 – ＋43.09(after dry-
ing, 0.5 g, N, N-dimethylformamide, 10 mL, 100 mm).
mL of the standard solution under the above operating con- pH <2.54> Dissolve 1.0 g of d-Chlorpheniramine Maleate in
ditions, the internal standard and chlorpheniramine are 100 mL of freshly boiled and cooled water: the pH of this so-
eluted in this order with the resolution between these peaks lution is between 4.0 and 5.0.
with 30 mL of the standard solution under the above operat- Purity (1) Clarity and color of solution—Dissolve 1.0 g
ing conditions, the relative standard deviation of the ratio of of d-Chlorpheniramine Maleate in 50 mL of water: the solu-
the peak area of chlorpheniramine to that of the internal tion is clear and colorless.
standard is not more than 1.0z. (2) Heavy metals <1.07>—Proceed with l.0 g of d-Chlor-
pheniramine Maleate according to Method 4 and perform
(3) Related substances—Dissolve 0.10 g of d-Chlor-
d-Chlorpheniramine Maleate pheniramine Maleate in 100 mL of the mobile phase, and use
d-クロルフェニラミンマレイン酸塩
ple solution, add the mobile phase to make exactly 100 mL.
Pipet 2 mL of this solution, add the mobile phase to make
exactly 20 mL, and use this solution as the standard solution.
method: the area of the peak other than maleic acid and d-
C16H19ClN2.C4H4O4: 390.86
chlorpheniramine obtained with the sample solution is not
(3S )-3-(4-Chlorophenyl)-N, N-dimethyl-3-pyridin-
larger than 2/3 times the peak area of d-chlorpheniramine
2-ylpropylamine monomaleate
obtained with the standard solution, and the total area of
[2438-32-6]
these peaks is not larger than the peak area of d-chlor-
pheniramine with the standard solution.
d-Chlorpheniramine Maleate, when dried, contains
not less than 99.0z and not more than 101.0z of d-
chlorpheniramine maleate (C16H19ClN2.C4H4O4).
length: 225 nm).
Description d-Chlorpheniramine Maleate occurs as a Column: A stainless steel column 3.9 mm in inside diame-
white, crystalline powder. ter and 30 cm in length, packed with octadecylsilanized silica
It is very soluble in water, in methanol and in acetic acid gel for liquid chromatography (10 mm in particle diameter).
(100), and freely soluble in N, N-dimethylformamide and in Column temperature: A constant temperature of about
ethanol (99.5). 259C.
It dissolves in dilute hydrochloric acid. Mobile phase: Dissolve 8.57 g of ammonium dihydrogen
phosphate and 1 mL of phosphoric acid in water to make
solution of d-Chlorpheniramine Maleate in 0.1 mol/L hy-
nitrile.
drochloric acid TS (3 in 100,000) as directed under Ultra-
Flow rate: Adjust so that the retention time of d-chlor-
692 Chlorpromazine Hydrochloride / Official Monographs JP XVII
retention time of d-chlorpheniramine, beginning after the 10 mL of 2,4,6-trinitrophenol TS, and allow to stand for 5
solvent peak. hours. Collect the resulting precipitate, wash with water,
System suitability— recrystallize from a small portion of acetone, and dry at
Test for required detectability: To exactly 2.5 mL of the 1059C for 1 hour: the crystals so obtained melt <2.60> be-
standard solution add the mobile phase to make exactly 25 tween 1759C and 1799C.
mL. Confirm that the peak area of d-chlorpheniramine ob- (3) Dissolve 0.5 g of Chlorpromazine Hydrochloride in 5
tained with 20 mL of this solution is equivalent to 7 to 13z mL of water, add 2 mL of ammonia TS, and heat on a water
of that obtained with 20 mL of the standard solution. bath for 5 minutes. Cool, filter, and render the filtrate acidic
System performance: When the procedure is run with 20 with dilute nitric acid: the solution responds to the Qualita-
mL of the standard solution under the above operating con- tive Tests <1.09> (2) for chloride.
Melting point <2.60> 194 – 1989
C
factor of the peak of d-chlorpheniramine are not less than
4000 and not more than 2.0, respectively. pH <2.54> Dissolve 1.0 g of Chlorpromazine Hydrochlo-
System repeatability: When the test is repeated 6 times ride in 20 mL of freshly boiled and cooled water, and meas-
with 20 mL of the standard solution under the above operat- ure within 10 minutes: the pH of this solution is between 4.0
ing conditions, the relative standard deviation of the peak and 5.0.
area of d-chlorpheniramine is not more than 4.0z.
Purity (1) Clarity and color of solution—A solution of
Loss on drying <2.41> Not more than 0.5z (1 g, 659C, 1.0 g of Chlorpromazine Hydrochloride in 20 mL of water,
4 hours). when observed within 10 minutes, is clear and colorless to
pale yellow.
Assay Weigh accurately about 0.3 g of d-Chlorphenira- promazine Hydrochloride according to Method 2, and per-
mine Maleate, previously dried, and dissolve in 20 mL of form the test. Prepare the control solution with 2.0 mL of
acetic acid (100). Titrate <2.50> with 0.1 mol/L perchloric Standard Lead Solution (not more than 20 ppm).
acid VS until the color of the solution changes from purple
through blue-green to green (indicator: 2 drops of crystal
2 hours).
violet TS). Perform a blank determination, and make any
necessary correction. Residue on ignition <2.44> Not more than 0.1z (1 g).
Each mL of 0.1 mol/L perchloric acid VS Assay Weigh accurately about 0.7 g of Chlorpromazine
＝ 19.54 mg of C16H19ClN2.C4H4O4 Hydrochloride, previously dried, dissolve in 50 mL of a mix-
ture of acetic anhydride and acetic acid (100) (7:3), and
metric titration). Perform a blank determination, and make
Chlorpromazine Hydrochloride Each mL of 0.1 mol/L perchloric acid VS

＝ 35.53 mg of C17H19ClN2S.HCl
クロルプロマジン塩酸塩
Chlorpromazine Hydrochloride
Injection
C17H19ClN2S.HCl: 355.33 クロルプロマジン塩酸塩注射液
3-(2-Chloro-10H-phenothiazin-10-yl)-N, N-
dimethylpropylamine monohydrochloride
Chlorpromazine Hydrochloride Injection is an
[69-09-0]
aqueous injection.
Chlorpromazine Hydrochloride, when dried, con-
105.0z of the labeled amount of chlorpromazine hy-
tains not less than 99.0z of chlorpromazine hydro-
drochloride (C17H19ClN2S.HCl: 355.33).
chloride (C17H19ClN2S.HCl).
Description Chlorpromazine Hydrochloride occurs as a
tions, with Chlorpromazine Hydrochloride.
white to pale yellow, crystalline powder. It is odorless, or has
a faint, characteristic odor. Description Chlorpromazine Hydrochloride Injection is a
It is very soluble in water, freely soluble in ethanol (95) clear, colorless or pale yellow liquid.
and in acetic acid (100), sparingly soluble in acetic anhy- pH: 4.0 – 6.5
dride, and practically insoluble in diethyl ether.
Identification (1) Proceed with a volume of Chlorproma-
zine Hydrochloride Injection, equivalent to 5 mg of Chlor-
Identification (1) To 5 mL of a solution of Chlorproma- promazine Hydrochloride, as directed in the Identification
zine Hydrochloride (1 in 1000) add 1 drop of iron (III) chlo- (1) under Chlorpromazine Hydrochloride.
ride TS: a red color develops. (2) Proceed with a volume of Chlorpromazine Hydro-
(2) Dissolve 0.1 g of Chlorpromazine Hydrochloride in chloride Injection, equivalent to 0.1 g of Chlorpromazine
20 mL of water and 3 drops of dilute hydrochloric acid, add Hydrochloride, as directed in the Identification (2) under
JP XVII Official Monographs / Chlorpromazine Hydrochloride Tablets 693
Chlorpromazine Hydrochloride. and ethanol (99.5) (1:1) so that each mL contains about 0.83
mg of chlorpromazine hydrochloride (C17H19ClN2S.HCl),
treat with the ultrasonic waves for 5 minutes, then shake
Foreign insoluble matter <6.06> Perform the test according vigorously for 20 minutes, and add the mixture of diluted
to Method 1: it meets the requirement. phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make
exactly V mL so that each mL contains about 0.5 mg of
chlorpromazine hydrochloride (C17H19ClN2S.HCl). Filter
ment.
Sterility <4.06> Perform the test according to the Mem- 0.45 mm. Discard the first 3 mL of the filtrate, pipet 2.5 mL
brane filtration method: it meets the requirement. of the subsequent filtrate, add exactly 5 mL of the internal
standard solution, then add the mixture of diluted phos-
Assay Transfer an exactly measured volume of Chlor-
phoric acid (1 in 500) and ethanol (99.5) (1:1) to make 25
promazine Hydrochloride Injection, equivalent to about
mL, and use this solution as the sample solution. Then,
0.15 g of chlorpromazine hydrochloride (C17H19ClN2S.HCl)
to a separator, add 30 mL of water and 10 mL of a solution
of sodium hydroxide (1 in 5), and extract with two 30-mL Amount (mg) of chlorpromazine hydrochloride
portions and three 20-mL portions of diethyl ether. Wash (C17H19ClN2S.HCl)
the combined diethyl ether extracts with successive 10-mL ＝ MS × QT/QS × V/50
portions of water until the last washing shows no red color
MS: Amount (mg) of chlorpromazine hydrochloride for
upon the addition of phenolphthalein TS. Concentrate the
assay taken
diethyl ether extracts on a water bath to 20 mL, add 5 g of
anhydrous sodium sulfate, allow to stand for 20 minutes, Internal standard solution—A solution of ethyl parahy-
and filter through a pledget of absorbent cotton. Wash with droxybenzoate in the mixture of diluted phosphoric acid (1
diethyl ether, combine the washings with the filtrate, and in 500) and ethanol (99.5) (1:1) (1 in 4500).
evaporate the diethyl ether on a water bath. Dissolve the
residue in 50 mL of acetone and 5 mL of acetic acid (100),
and titrate <2.50> with 0.05 mol/L perchloric acid VS until
mL of 2nd fluid for dissolution test as the dissolution me-
the color of the solution changes from red-purple to blue-
dium, the dissolution rate in 60 minutes of Chlorpromazine
purple (indicator: 3 drops of bromocresol green-methyl-
Hydrochloride Tablets is not less than 75z.
rosaniline chloride TS). Perform a blank determination in
Start the test with 1 tablet of Chlorpromazine Hydrochlo-
ride Tablets, withdraw not less than 20 mL of the medium at
Each mL of 0.05 mol/L perchloric acid VS the specified minute after starting the test, and filter through
＝ 17.77 mg of C17H19ClN2S.HCl a membrane filter with a pore size not exceeding 0.8 mm.
Containers and storage Containers—Hermetic containers,
subsequent filtrate, add the dissolution medium to make ex-
and colored containers may be used.
actly V? mL so that each mL contains about 5.6 mg of chlor-
promazine hydrochloride (C17H19ClN2S.HCl), and use this
about 90 mg of chlorpromazine hydrochloride for assay,
Chlorpromazine Hydrochloride previously dried at 1059C for 2 hours, dissolve in the dissolu-
Tablets tion medium to make exactly 200 mL. Pipet 5 mL of this so-
lution, add the dissolution medium to make exactly 100 mL,
クロルプロマジン塩酸塩錠 further pipet 5 mL of this solution, add the dissolution me-
dium to make exactly 20 mL, and use this solution as the
Chlorpromazine Hydrochloride Tablets contain not
of the sample solution and standard solution at 254 nm as di-
less than 93.0z and not more than 107.0z of the
labeled amount of chlorpromazine hydrochloride
(C17H19ClN2S.HCl: 355.33). Dissolution rate (z) with respect to labeled amount
of chlorpromazine hydrochloride (C17H19ClN2S.HCl)
＝ MS × AT/AS × V?/V × 1/C × 45/8
with Chlorpromazine Hydrochloride.
Identification (1) Shake a quantity of powdered Chlor-
assay taken
promazine Hydrochloride Tablets, equivalent to 0.2 g of
C: Labeled amount (mg) of chlorpromazine hydrochloride
Chlorpromazine Hydrochloride, with 40 mL of 0.1 mol/L
(C17H19ClN2S.HCl) in 1 tablet
hydrochloric acid TS, and filter. To 1 mL of the filtrate add
4 mL of water and 1 drop of iron (III) chloride TS: a red Assay Conduct this procedure without exposure to light,
color develops. using light-resistant vessels. Weigh accurately, and powder
(2) To 20 mL of the filtrate obtained in (1) add 10 mL of not less than 20 Chlorpromazine Hydrochloride Tablets.
2,4,6-trinitrophenol TS dropwise, and proceed as directed in Weigh accurately a portion of the powder, equivalent to
the Identification (2) under Chlorpromazine Hydrochloride. about 50 mg of chlorpromazine hydrochloride
(C17H19ClN2S.HCl), add 60 mL of a mixture of diluted phos-
phoric acid (1 in 500) and ethanol (99.5) (1:1), treat with
ultrasonic waves for 5 minutes, then shake vigorously for 20
minutes, and add the mixture of diluted phosphoric acid
Conduct this procedures using light-resistant vessels. To 1
(1 in 500) and ethanol (99.5) (1:1) to make exactly 100 mL.
tablet of Chlorpromazine Hydrochloride Tablets add an
Filter the solution through a membrane filter with a pore size
amount of a mixture of diluted phosphoric acid (1 in 500)
694 Chlorpropamide / Official Monographs JP XVII
not exceeding 0.45 mm, and discard the first 3 mL of the fil-
trate. To exactly 2.5 mL of the subsequent filtrate add ex- Chlorpropamide
actly 5 mL of the internal standard solution, then add the
mixture of diluted phosphoric acid (1 in 500) and ethanol クロルプロパミド
(99.5) (1:1) to make 25 mL, and use this solution as the sam-
chlorpromazine hydrochloride for assay, previously dried at
1059C for 2 hours, and dissolve in the mixture of diluted
phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make
exactly 100 mL. Pipet 5 mL of this solution, add exactly 5
C10H13ClN2O3S: 276.74
mL of the internal standard solution and the mixture of
4-Chloro-N-(propylcarbamoyl)benzenesulfonamide
diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to
[94-20-2]
Chlorpropamide, when dried, contains not less than
98.0z of chlorpropamide (C10H13ClN2O3S).
the ratios, QT and QS, of the peak area of chlorpromazine to Description Chlorpropamide occurs as white, crystals or
that of the internal standard. crystalline powder.
It is freely soluble in methanol and in acetone, soluble in
Amount (mg) of chlorpromazine hydrochloride
ethanol (95), and slightly soluble in diethyl ether, and practi-
(C17H19ClN2S.HCl) ＝ MS × QT/QS × 2
cally insoluble in water.
Identification (1) Dissolve 0.08 g of Chlorpropamide in
assay taken
50 mL of methanol. To 1 mL of the solution add 0.01 mol/L
Internal standard solution—A solution of ethyl parahy- hydrochloric acid TS to make 200 mL. Determine the ab-
droxybenzoate in a mixture of diluted phosphoric acid (1 in sorption spectrum of the solution as directed under Ultra-
500) and ethanol (99.5) (1:1) (1 in 4500). violet-visible Spectrophotometry <2.24>, and compare the
Operating conditions— spectrum with the Reference Spectrum: both spectra exhibit
Detector: An ultraviolet absorption photometer (wave- similar intensities of absorption at the same wavelengths.
length: 256 nm). (2) Determine the infrared absorption spectrum of
Column: A stainless steel column 4.6 mm in inside diame- Chlorpropamide, previously dried, as directed in the potas-
ter and 15 cm in length, packed with octadecylsilanized silica sium bromide disk method under Infrared Spectrophotome-
gel for liquid chromatography (5 mm in particle diameter). try <2.25>, and compare the spectrum with the Reference
Column temperature: A constant temperature of about Spectrum: both spectra exhibit similar intensities of absorp-
259 C. tion at the same wave numbers.
Mobile phase: A mixture of diluted 0.05 mol/L sodium di- (3) Perform the test with Chlorpropamide as directed
hydrogen phosphate TS (1 in 2) and acetonitrile (27:13). under Flame Coloration Test <1.04> (2): a green color ap-
Flow rate: Adjust so that the retention time of chlor- pears.
promazine is about 15 minutes.
Melting point <2.60> 127 – 1319
C
System performance: When the procedure is run with 10 Purity (1) Acidity—To 3.0 g Chlorpropamide add 150
mL of the standard solution under the above operating con- mL of water, and warm at 709 C for 5 minutes. Allow to
ditions, the internal standard and chlorpromazine are eluted stand in ice water for 1 hour, and filter. To 25 mL of the
in this order with the resolution between these peaks being filtrate add 2 drops of methyl red TS and 0.30 mL of 0.1
not less than 10. mol/L sodium hydroxide VS: a yellow color develops.
System repeatability: When the test is repeated 6 times (2) Chloride <1.03>—To 40 mL of the filtrate obtained in
with 10 mL of the standard solution under the above operat- (1) add 6 mL of dilute nitric acid and water to make 50 mL.
ing conditions, the relative standard deviation of the ratio of Perform the test using this solution as the test solution. Pre-
the peak area of chlorpromazine to that of the internal pare the control solution with 0.25 mL of 0.01 mol/L hydro-
standard is not more than 1.0z. chloric acid VS (not more than 0.011z).
propamide according to Method 2, and perform the test.
(5) Related substances—Dissolve 0.6 g of Chlor-
propamide in acetone to make exactly 10 mL, and use this
lution, add acetone to make exactly 300 mL, and use this so-
lution as the standard solution (1). Separately, dissolve 60
mg of 4-chlorobenzene sulfonamide in acetone to make ex-
(2). Perform the test with these solutions as directed under
JP XVII Official Monographs / Chlorpropamide Tablets 695
Thin-layer Chromatography <2.03>. Spot 5 mL each of the um, the dissolution rate in 45 minutes of Chlorpropamide
sample solution and standard solutions (1) and (2) on a plate Tablets is not less than 70z.
of silica gel for thin-layer chromatography. Develop the Start the test with 1 tablet of Chlorpropamide Tablets,
plate with a mixture of cyclohexane, 3-methyl-1-butanol, withdraw not less than 20 mL of the medium at the specified
methanol and ammonia solution (28) (15:10:5:1) to a dis- minute after starting the test, and filter through a membrane
tance of about 10 cm, and air-dry the plate. After drying the filter with a pore size not exceeding 0.8 mm. Discard the first
plate at 1009C for 1 hour, spray evenly sodium hypochlorite 10 mL of the filtrate, pipet V mL of the subsequent filtrate,
TS on the plate, and air-dry for 15 minutes. Then spray add the dissolution medium to make exactly V? mL so that
evenly potassium iodide-starch TS on the plate: the spot each mL contains about 10 mg of chlorpropamide
from the sample solution equivalent to the spot from the (C10H13ClN2O3S), and use this solution as the sample solu-
standard solution (2) is not more intense than the spot from tion. Separately, weigh accurately about 50 mg of chlor-
the standard solution (2), and the spots other than the spot propamide for assay, previously dried at 1059 C for 3 hours,
mentioned above and other than the principal spot is not dissolve in 10 mL of methanol, and add water to make ex-
more intense than the spot from the standard solution (1). actly 50 mL. Pipet 1 mL of this solution, add the dissolution
3 hours).
of the sample solution and standard solution at 232 nm as di-
Residue on ignition <2.44> Not more than 0.2z (1 g). rected under Ultraviolet-visible Spectrophotometry <2.24>.
Assay Weigh accurately about 0.5 g of Chlorpropamide, Dissolution rate (z) with respect to the labeled amount
previously dried, dissolve in 30 mL of neutralized ethanol, of chlorpropamide (C10H13ClN2O3S)
and add 20 mL of water. Titrate <2.50> with 0.1 mol/L so- ＝ MS × AT/AS × V?/V × 1/C × 18
dium hydroxide VS (indicator: 3 drops of phenolphthalein
MS: Amount (mg) of chlorpropamide for assay taken
TS).
C: Labeled amount (mg) of chlorpropamide
Each mL of 0.1 mol/L sodium hydroxide VS (C10H13ClN2O3S) in 1 tablet
＝ 27.67 mg of C10H13ClN2O3S
Containers and storage Containers—Well-closed contain- Chlorpropamide Tablets. Weigh accurately a quantity of the
ers. powder, equivalent to about 50 mg of chlorpropamide
(C10H13ClN2O3S), add 75 mL of the mobile phase, shake for
10 minutes, and add the mobile phase to make exactly 100
Chlorpropamide Tablets mL. Centrifuge this solution, pipet 10 mL of the supernatant
liquid, add the mobile phase to make exactly 100 mL, and
クロルプロパミド錠 use this solution as the sample solution. Separately, weigh
accurately about 50 mg of chlorpropamide for assay, previ-
ously dried at 1059C for 3 hours, dissolve in the mobile
Chlorpropamide Tablets contain not less than
amount of chlorpropamide (C10H13ClN2O3S: 276.74).
Method of preparation Prepare as directed under Tablets, actly 20 mL each of the sample solution and standard solu-
with Chlorpropamide. tion as directed under Liquid Chromatography <2.01> ac-
cording to the following conditions. Determine the peak
Identification Take a quantity of powdered Chlor-
areas, AT and AS, of chlorpropamide in each solution.
propamide Tablets, equivalent to 0.08 g of Chlorpropamide,
add 50 mL of methanol, shake, and filter. To 1 mL of the Amount (mg) of chlorpropamide (C10H13ClN2O3S)
filtrate add 0.01 mol/L hydrochloric acid TS to make 200 ＝ M S × AT / AS
<2.24>: it exhibits a maximum between 231 nm and 235 nm. Operating conditions—
length: 240 nm).
To 1 tablet of Chlorpropamide Tablets add 75 mL of the
mobile phase, treat with the ultrasonic waves for 20 minutes
with occasional strong shaking, then add the mobile phase to
259C.
of Chlorpropamide. Centrifuge the solution, pipet 2 mL of
100) and acetonitrile (1:1).
the supernatant liquid, add the mobile phase to make exactly
Flow rate: Adjust so that the retention time of chlor-
100 mL, and use this solution as the sample solution. Then,
propamide is about 5 minutes.
Amount (mg) of chlorpropamide (C10H13ClN2O3S) System performance: When the procedure is run with 20
＝ MS × AT/AS × V/20 mL of the standard solution under the above operating con-
factor of the peak of chlorpropamide are not less than 1500
Dissolution <6.10> When the test is performed at 50 revolu- and not more than 1.5, respectively.
tions per minute according to the Paddle method, using 900 System repeatability: When the test is repeated 6 times
mL of 2nd fluid for dissolution test as the dissolution medi- with 20 mL of the standard solution under the above operat-
696 Cholecalciferol / Official Monographs JP XVII
ing conditions, the relative standard deviation of the peak Assay Proceed with the operation avoiding contact with air
area of chlorpropamide is not more than 1.5z. or other oxidizing agents and using light-resistant containers.
Dissolve separately about 30 mg each of Cholecalciferol and
Cholecalciferol RS, accurately weighed, in isooctane to
ers.
make exactly 50 mL. Pipet 10 mL each of these solutions,
add 3 mL each of the internal standard solution, then add
Cholecalciferol the sample solution and the standard solution. Perform the
Vitamin D3 lution as directed under Liquid Chromatography <2.01> ac-
コレカルシフェロール
QT and QS, of the peak area of cholecalciferol to that of the
internal standard.
Amount (mg) of cholecalciferol (C27H44O ＝ MS × QT/QS)
MS: Amount (mg) of Cholecalciferol RS taken
late in isooctane (1 in 100).
length: 254 nm).
Column: A stainless steel column about 4 mm in inside di-
C27H44O: 384.64 ameter and 10 to 30 cm in length, packed with silica gel for
(3S,5Z,7E )-9,10-Secocholesta-5,7,10(19)-trien-3-ol liquid chromatography (5 to 10 mm in particle diameter).
[67-97-0] Column temperature: Ordinary temperature.
Mobile phase: A mixture of hexane and n-amylalcohol
Cholecalciferol contains not less than 97.0z and (997:3).
not more than 103.0z of cholecalciferol (C27H44O). Flow rate: Adjust so that the retention time of cholecal-
ciferol is about 25 minutes.
Description Cholecalciferol occurs as white crystals. It is
Selection of column: Dissolve 15 mg of Cholecalciferol RS
odorless.
in 25 mL of isooctane. Transfer this solution to a flask, heat
It is freely soluble in ethanol (95), in chloroform, in
under a reflux condenser in an oil bath for 2 hours, and cool
diethyl ether and in isooctane, and practically insoluble in
to room temperature rapidly. Transfer this solution to a
water.
quartz test tube, and irradiate under a short-wave lamp
It is affected by air and by light.
(main wavelength: 254 nm) and a long-wave lamp (main
Melting point: 84 – 889C Transfer Cholecalciferol to a
wavelength: 365 nm) for 3 hours. To this solution add the
capillary tube, and dry for 3 hours in a desiccator (in vacu-
mobile phase to make 50 mL. Proceed with 10 mL of this so-
um at a pressure not exceeding 2.67 kPa). Immediately
lution under the above operating conditions. Use a column
fireseal the capillary tube, put it in a bath fluid, previously
with the relative retention time of previtamin D3, trans-vita-
heated to a temperature about 109C below the expected
min D3 and tachysterol D3 to cholecalciferol being about 0.5,
melting point, and heat at a rate of rise of about 39C per
about 0.6 and about 1.1, respectively, and with resolution
minute, and read the melting point.
between previtamin D3 and trans-vitamin D3, and that be-
Identification (1) Dissolve 0.5 mg of Cholecalciferol in 5 tween cholecalciferol and tachysterol D3 being not less than
mL of chloroform, add 0.3 mL of acetic anhydride and 0.1 1.0.
mL of sulfuric acid, and shake: a red color is produced, and
rapidly changes through purple and blue to green.
Storage—Light-resistant, under nitrogen atmosphere, and
in a cold place.
Cholecalciferol as directed in the potassium bromide disk
trum of Cholecalciferol RS: both spectra exhibit similar in- Cholera Vaccine
コレラワクチン
Absorbance <2.24> E 11zcm (265 nm): 450 – 490 (10 mg,
ethanol (95), 1000 mL).
Cholera Vaccine is a liquid for injection containing
Optical rotation <2.49> [a]20 D ＋103 – ＋1129 (50 mg, inactivated Vibrio cholerae of the Ogawa and Inaba
ethanol (95), 10 mL, 100 mm). Prepare the solution without strains.
delay, using Cholecalciferol from a container opened not Monotypic products may be manufactured, if neces-
longer than 30 minutes, previously, and determine the rota- sary.
tion within 30 minutes after the solution has been prepared. It conforms to the requirements of Cholera Vaccine
in the Minimum Requirements for Biological
Purity 7-Dehydrocholesterol—Dissolve 10 mg of Cholecal-
Products.
ciferol in 2.0 mL of diluted ethanol (95) (9 in 10), add a solu-
tion prepared by dissolving 20 mg of digitonin in 2.0 mL of Description Cholera Vaccine is a white-turbid liquid.
diluted ethanol (95) (9 in 10), and allow the mixture to stand
for 18 hours: no precipitate is formed.
JP XVII Official Monographs / Cibenzoline Succinate 697
Cholesterol Cibenzoline Succinate

コレステロールシベンゾリンコハク酸塩
C18H18N2.C4H6O4: 380.44
C27H46O: 386.65 2-[(1RS )-2,2-Diphenylcyclopropan-1-yl]-4,5-dihydro-1H-
Cholest-5-en-3b-ol imidazole monosuccinate
[57-88-5] [100678-32-8]
Description Cholesterol occurs as white to pale yellow crys-
Cibenzoline Succinate, when dried, contains not
tals or granules. It is odorless, or has a slight odor. It is
less than 98.5z and not more than 101.0z of ciben-
tasteless.
zoline succinate (C18H18N2.C4H6O4).
It is freely soluble in chloroform and in diethyl ether, solu-
ble in 1,4-dioxane, sparingly soluble in ethanol (99.5), and Description Cibenzoline Succinate occurs as a white crys-
practically insoluble in water. talline powder.
It gradually changes to a yellow to light yellow-brown It is freely soluble in methanol and in acetic acid (100),
color by light. and sparingly soluble in water and in ethanol (99.5).
A solution of Cibenzoline Succinate in methanol (1 in 10)
Identification (1) Dissolve 0.01 g of Cholesterol in 1 mL
shows no optical rotation.
of chloroform, add 1 mL of sulfuric acid, and shake: a red
color develops in the chloroform layer, and the sulfuric acid Identification (1) Determine the absorption spectrum of a
layer shows a green fluorescence. solution of Cibenzoline Succinate (1 in 50,000) as directed
(2) Dissolve 5 mg of Cholesterol in 2 mL of chloroform, under Ultraviolet-visible Spectrophotometry <2.24>, and
add 1 mL of acetic anhydride and 1 drop of sulfuric acid, compare the spectrum with the Reference Spectrum: both
and shake: a red color is produced, and it changes to green spectra exhibit similar intensities of absorption at the same
through blue. wavelengths.
D : －34 – －389 (after drying,
Cibenzoline Succinate as directed in the paste method under
0.2 g, 1,4-dioxane, 10 mL, 100 mm).
Melting point <2.60> 147 – 1509C trum with the Reference Spectrum: both spectra exhibit simi-
Purity (1) Clarity of solution—Place 0.5 g of Cholesterol
(3) Shake 0.4 g of Cibenzoline Succinate with 2.5 mL of
in a glass-stoppered flask, dissolve in 50 mL of warm ethanol
sodium hydroxide TS and 5 mL of ethyl acetate, allow to
(95), and allow to stand at room temperature for 2 hours: no
stand, and to 1 mL of the water layer so obtained add 0.5
turbidity or deposit is produced.
mL of 1 mol/L hydrochloric acid TS and 0.5 mL of iron
(2) Acidity—Place 1.0 g of Cholesterol in a flask, dis-
(III) chloride TS: a blown precipitate is formed.
solve in 10 mL of diethyl ether, add 10.0 mL of 0.1 mol/L
sodium hydroxide VS, and shake for 1 minute. Expel the Melting point <2.60> 163 – 1679C
diethyl ether, and boil for 5 minutes. Cool, add 10 mL of
pH <2.54> Dissolve 0.20 g of Cibenzoline Succinate in 10
water, and titrate <2.50> with 0.05 mol/L sulfuric acid VS
(indicator: 2 drops of phenolphthalein TS). Perform a blank
determination in the same manner. Purity (1) Clarity and color of solution—A solution ob-
The volume of 0.1 mol/L sodium hydroxide VS consumed tained by dissolving 0.20 g of Cibenzoline Succinate in 10
is not more than 0.30 mL. mL of water is clear and colorless.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Cibenzo-
line Succinate according to Method 2, and perform the test.
um, 609C, 4 hours).
Residue on ignition <2.44> Not more than 0.1z (1 g). Solution (not more than 20 ppm).
of Cibenzoline Succinate according to Method 3, using a so-
lution of magnesium nitrate hexahydrate in ethanol (95) (1 in
25), and perform the test (not more than 2 ppm).
(4) Related substances—Dissolve 0.10 g of Cibenzoline
Succinate in 10 mL of methanol, and use this solution as the
sample solution. Pipet 1 mL of the sample solution, and add
methanol to make exactly 100 mL. Pipet 5 mL and 2 mL of
this solution, add methanol to make them exactly 10 mL,
and use these solutions as the standard solution (1) and the
standard solution (2). Perform the test with these solutions
10 mL each of the sample solution and standard solutions (1)
698 Cibenzoline Succinate Tablets / Official Monographs JP XVII
and (2) on a plate of silica gel with fluorescent indicator for Amount (mg) of cibenzoline succinate (C18H18N2.C4H6O4)
thin-layer chromatography. Develop the plate with a mixture ＝ MS × QT/QS × C/100
of ethyl acetate, methanol and ammonia solution (28)
MS: Amount (mg) of cibenzoline succinate for assay taken
(20:3:2) to a distance of about 10 cm, air-dry the plate, and
C: Labeled amount (mg) of cibenzoline succinate
dry more at 809C for 30 minutes. After cooling, examine
(C18H18N2.C4H6O4) in 1 tablet
under ultraviolet light (main wavelength: 254 nm): the spot
other than the principal spot obtained with the sample solu- Internal standard solution—Dissolve 0.1 g of 2-ethylhexyl
tion is not more intense than the spot obtained with the parahydroxybenzoate in methanol to make 100 mL.
standard solution (1). Allow the plate to stand for 30
minutes in iodine vapor: the spot other than the principal
spot obtained with the sample solution is not more intense
than the spot obtained with the standard solution (1), and
in 15 minutes of Cibenzoline Succinate Tablets is not less
the spot, which is more intense than the spot with the stand-
than 80z.
ard solution (2), is not more than two.
Start the test with 1 tablet of Cibenzoline Succinate
Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, Tablets, withdraw not less than 20 mL of the medium at the
2 hours). specified minute after starting the test, and filter through a
Assay Weigh accurately about 0.4 g of Cibenzoline Suc- subsequent filtrate, add water to make exactly V? mL so that
cinate, previously dried, dissolve in 50 mL of acetic acid each mL contains about 11 mg of cibenzoline succinate
(100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (C18H18N2.C4H6O4), and use this solution as the sample solu-
until the color of the solution changes from violet to blue- tion. Separately, weigh accurately about 28 mg of cibenzo-
green through blue (indicator: 2 drops of crystal violet TS). line succinate for assay, previously dried at 1059C for 2
Perform a blank determination in the same manner, and hours, and dissolve in water to make exactly 100 mL. Pipet 2
make any necessary correction. mL of this solution, add water to make exactly 50 mL, and
＝ 38.04 mg of C18H18N2.C4H6O4
Containers and storage Containers—Tight containers. trophotometry <2.24>.
of cibenzoline succinate (C18H18N2.C4H6O4)
Cibenzoline Succinate Tablets ＝ MS × AT/AS × V?/V × 1/C × 36
シベンゾリンコハク酸塩錠 MS: Amount (mg) of cibenzoline succinate for assay taken
C: Labeled amount (mg) of cibenzoline succinate
(C18H18N2.C4H6O4) in 1 tablet
Cibenzoline Succinate Tablets contain not less than
95.0z and not more than 105.0z of the labeled Assay Weigh accurately not less than 20 Cibenzoline Suc-
amount of cibenzoline succinate (C18H18N2.C4H6O4: cinate Tablets, and powder. Weigh accurately a portion of
380.44). the powder, equivalent to about 0.1 g of cibenzoline suc-
cinate (C18H18N2.C4H6O4), add 10 mL of water, shake, and
add 40 mL of methanol and exactly 10 mL of the internal
with Cibenzoline Succinate.
standard solution. Shake for 20 minutes, add methanol to
Identification To a quantity of powdered Cibenzoline Suc- make 100 mL, centrifuge, and use the supernatant liquid as
cinate Tablets, equivalent to 50 mg of Cibenzoline Succinate, the sample solution. Separately, weigh accurately about 0.1 g
add 100 mL of water, shake for 10 minutes, and centrifuge. of cibenzoline succinate for assay, previously dried at 1059
C
To 2 mL of the supernatant liquid add water to make 50 mL, for 2 hours, add 10 mL of water and 40 mL of methanol to
and determine the absorption spectrum of this solution as di- dissolve, then add exactly 10 mL of the internal standard so-
rected under Ultraviolet-visible Spectrophotometry <2.24>: it lution and methanol to make 100 mL, and use this solution
exhibits a maximum between 221 nm and 225 nm. as the standard solution. Perform the test with 5 mL each of
area of cibenzoline to that of the internal standard.
To 1 tablet of Cibenzoline Succinate Tablets add a suitable
amount of water so that each mL contains about 10 mg of Amount (mg) of cibenzoline succinate (C18H18N2.C4H6O4)
cibenzoline succinate (C18H18N2.C4H6O4), and allow standing ＝ M S × Q T / QS
for 10 minutes while occasional shaking. To this solution
MS: Amount (mg) of cibenzoline succinate for assay taken
add methanol so that each mL contains about 2 mg of ciben-
zoline succinate (C18H18N2.C4H6O4), add exactly 1 mL of the Internal standard solution—Dissolve 0.1 g of 2-ethylhexyl
internal standard solution per 10 mg of cibenzoline succinate parahydroxybenzoate in methanol to make 100 mL.
(C18H18N2.C4H6O4), then add methanol so that each mL Operating conditions—
contains about 1 mg of cibenzoline succinate Detector: An ultraviolet absorption photometer (wave-
(C18H18N2.C4H6O4). Centrifuge the solution, and use the length: 254 nm).
supernatant liquid as the sample solution. Then, proceed as Column: A stainless steel column 4.6 mm in inside diame-
directed in the Assay. ter and 5 cm in length, packed with octadecylsilanized silica
JP XVII Official Monographs / Ciclacillin 699
Column temperature: A constant temperature of about Assay Weigh accurately an amount of Ciclacillin and
259 C. Ciclacillin RS, equivalent to about 50 mg (potency), dissolve
Mobile phase: Dissolve 2.67 g of sodium di-2-ethylhexyl each in a suitable amount of the mobile phase, add exactly 5
sulfosuccinate in 2000 mL of a mixture of water, acetonitrile mL of the internal standard solution and the mobile phase to
and diluted phosphoric acid (1 in 10) (1000:1000:1). make 50 mL, and use these solutions as the sample solution
Flow rate: Adjust so that the retention time of cibenzoline and standard solution. Perform the test with 10 mL each of
is about 3 minutes. the sample solution and standard solution as directed under
System suitability— Liquid Chromatography <2.01> according to the following
System performance: When the procedure is run with 5 mL conditions, and calculate the ratios, QT and QS, of the peak
of the standard solution under the above operating condi- area of ciclacillin to that of the internal standard.
tions, cibenzoline and the internal standard are eluted in this
Amount [ mg (potency)] of ciclacillin (C15H23N3O4S)
＝ MS × QT/QS × 1000
than 6.
System repeatability: When the test is repeated 6 times MS: Amount [mg (potency)] of Ciclacillin RS taken
Internal standard solution—A solution of orcin in the mo-
bile phase (1 in 500).
peak area of cibenzoline to that of the internal standard is
not more than 1.0z.
Containers and storage Containers—Tight containers. length: 254 nm).
Ciclacillin for liquid chromatography (5 mm in particle diameter).
シクラシリン 259C.
Mobile phase: Dissolve 0.771 g of ammonium acetate in
about 900 mL of water, adjust the pH to 4.0 with acetic acid
Flow rate: Adjust so that the retention time of ciclacillin is
about 4 minutes.
C15H23N3O4S: 341.43 System suitability—
(2S,5R,6R)-6-[(1-Aminocyclohexanecarbonyl)amino]- System performance: When the procedure is run with 10
3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2- mL of the standard solution under the above operating con-
carboxylic acid ditions, ciclacillin and the internal standard are eluted in this
[3485-14-1] order with the resolution between these peaks being not less
than 8.
Ciclacillin contains not less than 920 mg (potency) System repeatability: When the test is repeated 6 times
and not more than 1010 mg (potency) per mg, calcu- with 10 mL of the standard solution under the above operat-
lated on the anhydrous basis. The potency of Ciclacil- ing conditions, the relative standard deviation of the ratios
lin is expressed as mass (potency) of ciclacillin of the peak areas of ciclacillin to that of the internal stand-
(C15H23N3O4S). ard is not more than 1.0z.
Description Ciclacillin occurs as white to light yellowish Containers and storage Containers—Tight containers.
nol, and practically insoluble in acetonitrile and in ethanol
(99.5).
of Ciclacillin as directed in the potassium bromide disk
trum of Ciclacillin RS: both spectra exhibit similar intensities
D : ＋300 – ＋3159 (2 g, water,
100 mL, 100 mm).
Ciclacillin according to Method 2, and perform the test. Pre-
of Ciclacillin according to Method 3, and perform the test
700 Ciclosporin / Official Monographs JP XVII
der Liquid Chromatography <2.01> according to the follow-
Ciclosporin ing conditions. Determine each peak area of both solutions
by the automatic integration method: the area of the peak
Ciclosporin A other than the ciclosporin from the sample solution is not
larger than 7/10 times the peak area of ciclosporin from the
シクロスポリン standard solution, and the total area of all peaks other than
the ciclosporin from the sample solution is not larger than
1.5 times the peak area of ciclosporin from the standard so-
lution.
the Assay.
retention time of ciclosporin, beginning after the solvent
peak.
C62H111N11O12: 1202.61
cyclo{-[(2S,3R,4R,6E )-3-Hydroxy-4-methyl-2-
methylaminooct-6-enoyl]-L-2-aminobutanoyl-
standard solution add a mixture of water and acetonitrile
N-methylglycyl-N-methyl-L-leucyl-L-valyl-N-methyl-
L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-
ciclosporin obtained from 20 mL of this solution is equiva-
L-leucyl-N-methyl-L-valyl-}
lent to 7 to 13z of that of ciclosporin obtained from 20 mL
[59865-13-3]
of the standard solution.
Ciclosporin contains not less than 98.5z and not
more than 101.5z of ciclosporin (C62H111N11O12), cal-
area of ciclosporin is not more than 3.0z.
Description Ciclosporin occurs as a white powder.
It is very soluble in acetonitrile, in methanol and in
at a pressure not exceeding 0.67 kPa, 609C, 3 hours).
ethanol (95), freely soluble in diethyl ether, and practically
insoluble in water. Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Identification Determine the infrared absorption spectrum Assay Weigh accurately about 30 mg each of Ciclosporin
of Ciclosporin as directed in the potassium bromide disk and Ciclosporin RS (separately determine the loss on drying
method under Infrared Spectrophotometry <2.25>, and com- <2.41> under the same conditions as Ciclosporin), and dis-
pare the spectrum with the Reference Spectrum or the spec- solve each in a mixture of water and acetonitrile (1:1) to
trum of Ciclosporin RS: both spectra exhibit similar intensi- make exactly 25 mL, and use these solutions as the sample
ties of absorption at the same wave numbers. solution and standard solution. Perform the test with exactly
Optical rotation <2.49> [a]20D : －185 – －1939(0.1 g calcu-
lated on the dried basis, methanol, 20 mL, 100 mm).
the following conditions, and determine the peak areas, AT
Purity (1) Clarity and color of solution—Dissolve 1.0 g and AS, of ciclosporin in each solution.
of Ciclosporin in 10 mL of ethanol (95): the solution is clear,
Amount (mg) of ciclosporin (C62H111N11O12)
and has no more color than the following control solutions
＝ M S × A T / AS
(1), (2) or (3).
Control solution (1): To exactly 3.0 mL of Iron (III) Chlo- MS: Amount (mg) of Ciclosporin RS taken, calculated on
ride CS and exactly 0.8 mL of Cobalt (II) Chloride CS add the dried basis
diluted hydrochloric acid (1 in 40) to make exactly 100 mL.
Control solution (2): To exactly 3.0 mL of Iron (III) Chlo-
ride CS, exactly 1.3 mL of Cobalt (II) Chloride CS and ex-
length: 210 nm).
actly 0.5 mL of Copper (II) Sulfate CS add diluted hydro-
chloric acid (1 in 40) to make exactly 100 mL.
Control solution (3): To exactly 0.5 mL of Iron (III) Chlo-
for liquid chromatography (5 mm in particle diameter). Con-
ride CS and exactly 1.0 mL of Cobalt (II) Chloride CS add
nect the sample injection port and the column with a stain-
diluted hydrochloric acid (1 in 40) to make exactly 100 mL.
less steel tube 0.3 mm in inside diameter and 1 m in length.
Ciclosporin according to Method 2, and perform the test.
809C (including the sample injection port and the connecting
tube).
Mobile phase: A mixture of water, acetonitrile, tert-butyl
methyl ether and phosphoric acid (520:430:50:1).
Flow rate: Adjust so that the retention time of ciclosporin
sample solution, add a mixture of water and acetonitrile
System performance: Dissolve 3 mg of Ciclosporin U in
of the sample solution and standard soluton as directed un-
2.5 mL of a mixture of water and acetonitrile (1:1), and add
JP XVII Official Monographs / Cilastatin Sodium 701
2.5 mL of the standard solution. When the procedure is run nitric acid. Prepare the control solution with 2.0 mL of
with 20 mL of this solution under the above operating condi- Standard Lead Solution (not more than 20 ppm).
tions, ciclosporin U and ciclosporin are eluted in this order (3) Arsenic <1.11>—To 2.0 g of Cilastatin Sodium add 5
with the resolution between these peaks being not less than mL of nitric acid and 1 mL of sulfuric acid, and heat care-
1.2. fully until white fumes are evolved. After cooling, heat with
System repeatability: When the test is repeated 6 times two 2-mL portions of nitric acid, then heat with several
with 20 mL of the standard solution under the above operat- 2-mL portions of hydrogen peroxide (30) until a colorless or
ing conditions, the relative standard deviation of the peak pale yellow solution is obtained. After cooling, heat again
area of ciclosporin is not more than 1.0z. until white fumes are evolved. After cooling, add water to
make 5 mL, and perform the test with this solution as the
test solution: it shows no more color than the following color
standard.
Color standard: Prepare a solution according to the above
procedure without using Cilastatin Sodium, add exactly 2
Cilastatin Sodium mL of Standard Arsenic Solution, and perform the test in
the same manner as the test solution (not more than 1 ppm).
シラスタチンナトリウム
(4) Related substances—Dissolve about 40 mg of Cilasta-
tin Sodium in 25 mL of water, and use this solution as the
of the sample solution and standard solution as directed un-
der Liquid Chromatography <2.01> according to the follow-
C16H25N2NaO5S: 380.43 ing conditions, and determine each peak area by the auto-
Monosodium (2Z )-7-{[(2R)-2-amino- matic integration method: the area of the peak other than
2-carboxyethyl]sulfanyl}-2-({[(1S )-2,2- cilastatin from the sample solution is not larger than 1/6
dimethylcyclopropyl]carbonyl}amino)hept-2-enoate times the peak area of cilastatin from the standard solution,
[81129-83-1] and the total area of the peaks other than the peak of cilasta-
tin from the sample solution is not larger than the peak area
Cilastatin Sodium contains not less than 98.0z of cilastatin from the standard solution.
and not more than 101.0z of cilastatin sodium Operating conditions—
(C16H25N2NaO5S), calculated on the anhydrous and Detector: An ultraviolet absorption photometer (wave-
residual solvent-free basis. length: 210 nm).
Description Cilastatin Sodium occurs as a white to pale yel-
It is very soluble in water, freely soluble in methanol, and
slightly soluble in ethanol (99.5).
509C.
It is hygroscopic.
Mobile phase A: A mixture of diluted phosphoric acid
Identification (1) Determine the infrared absorption spec- (1 in 1000) and acetonitrile (7:3).
trum of Cilastatin Sodium as directed in the potassium bro- Mobile phase B: Diluted phosphoric acid (1 in 1000).
mide disk method under Infrared Spectrophotometry <2.25>, Flowing of mobile phase: Control the gradient by mixing
and compare the spectrum with the Reference Spectrum: the mobile phases A and B as directed in the following table.
same wave numbers. Time after injection Mobile phase A Mobile phase B
(2) A solution of Cilastatin Sodium (1 in 10) responds to of sample (min) (volz) (volz)
the Qualitative Tests <1.09> for sodium salt.
0 – 30 15 → 100 85 → 0
D : ＋41.5 – ＋44.59(0.1 g calcu-
30 – 40 100 0
lated on the anhydrous and residual solvent-free basis, a so-
lution of hydrochloric acid in methanol (9 in 1000), 10 mL,
100 mL). Flow rate: 2.0 mL per minute.
Time span of measurement: For 40 minutes.
pH <2.54> The pH of a solution prepared by dissolving System suitability—
1.0 g of Cilastatin Sodium in 100 mL of water is between 6.5 Test for required detectability: Pipet 1 mL of the standard
and 7.5. solution, and add water to make exactly 30 mL. Confirm
Purity (1) Clarity and color of solution—Dissolve 1.0 g that the peak area of cilastatin obtained with 20 mL of this
of Cilastatin Sodium in 100 mL of water: the solution is solution is equivalent to 2.3 to 4.5z of that obtained with 20
clear and the solution has no more color than the following mL of the standard solution.
control solution. System performance: When the procedure is run with 20
Control solution: To a mixture of 2.4 mL of Iron (III) mL of the standard solution under the above operating con-
Chloride CS and 0.6 mL of Cobalt (II) Chloride CS add ditions, the retention time of cilastatin is about 20 minutes,
water to make 10 mL, pipet 5 mL of this solution, and add and the number of theoretical plates and the symmetry fac-
water to make exactly 100 mL. tor of the peak of cilastatin are not less than 10,000 and not
(2) Heavy metals <1.07>—Proceed with 1.0 g of Cilasta- more than 2.5, respectively.
tin Sodium according to Method 2, and perform the test. System repeatability: When the test is repeated 3 times
After carbonization, add 0.5 mL of sulfuric acid instead of with 20 mL of the standard solution under the above operat-
702 Cilazapril Hydrate / Official Monographs JP XVII
ing conditions, the relative standard deviation of the peak tiometric titration).
area of cilastatin is not more than 2.0z.
(5) Residual solvents <2.46>—Weigh accurately about
＝ 19.02 mg of C16H25N2NaO5S
0.2 g of Cilastatin Sodium, add exactly 2 mL of the internal
standard solution, dissolve in water to make 10 mL, and use Containers and storage Containers—Tight containers.
this solution as the sample solution. Separately, measure ex- Storage—In a cold place.
actly 2 mL of acetone, 0.5 mL of methanol and 0.5 mL of
mesityl oxide, and add water to make exactly 1000 mL. Pipet
2 mL of this solution, add exactly 2 mL of the internal stand- Cilazapril Hydrate
ard solution, add water to make 10 mL, and use this solution
as the standard solution. Perform the test with 2 mL each of シラザプリル水和物
Gas Chromatography <2.02> according to the following con-
ditions. Calculate the ratios of the peak areas of acetone,
methanol and mesityl oxide and to the peak area of the inter-
nal standard, QTa and QSa, QTb and QSb, QTc and QSc, and
calculate the amounts of acetone, methanol and mesityl
oxide by the following equation: they are not more than C22H31N3O5.H2O: 435.51
1.0z, not more 0.5z and not more than 0.4z, respectively. (1S,9S )-9-[(1S )-(1-Ethoxycarbonyl-
3-phenylpropyl)amino]-10-oxooctahydro-
Amount (z) of acetone (CH3COCH3)
6H-pyridazino[1,2-a][1,2]diazepine-1-carboxylic acid
＝ 1/MT × QTa/QSa × 400 × 0.79
monohydrate
Amount (z) of methanol (CH3OH) [92077-78-6]
＝ 1/MT × QTb/QSb × 100 × 0.79
Cilazapril Hydrate contains not less than 98.5z and
Amount (z) of mesityl oxide (CH3COCH ＝ C(CH3)2)
＝ 1/MT × QTc/QSc × 100 × 0.86
not more than 101.0z of cilazapril (C22H31N3O5:
417.50), calculated on the anhydrous basis.
MT: Amount (mg) of Cilastatin Sodium taken
Description Cilazapril Hydrate occurs as white to yellowish
0.79: Density (g/mL) of acetone and methanol
0.86: Density (g/mL) of mesityl oxide
It is very soluble in methanol, freely soluble in ethanol
Internal standard solution—To 0.5 mL of 1-propanol add (99.5) and in acetic acid (100), and slightly soluble in water.
water to make 1000 mL. It gradually turns yellow on exposure to light.
Operating conditions— Melting point: about 1019 C (with decomposition).
Identification (1) To 4 mL of a solution of Cilazapril Hy-
Column: A glass column 3.2 mm in inside diameter and
drate (1 in 1000) add 2 mL of Dragendorff's TS: an orange
2.1 m in length, packed with tetrafluoroethylene polymer for
gas chromatography (250 – 420 mm) coated with polyethy-
lene glycol 20 M for gas chromatography at the ratio of
Cilazapril Hydrate as directed in the potassium bromide disk
10z.
709 C.
numbers.
standard is about 5 minutes. Optical rotation <2.49> [a]20
D : －53 – －589(0.2 g calculated
Time span of measurement: About 3 times as long as the on the anhydrous basis, methanol, 20 mL, 100 mm).
retention time of the internal standard.
Purity (1) Chloride <1.03>—Perform the test using 1.0 g
of Cilazapril Hydrate. Prepare the control solution with
0.009z).
tions, acetone, methanol, 1-propanol and mesityl oxide are
(2) Sulfate <1.14>—Dissolve 1.0 g of Cilazapril Hydrate
eluted in this order, and these peaks completely separate
in 40 mL of water and 1.5 mL of dilute hydrochloric acid,
each other.
and add water to make 50 mL. Perform the test using this
solution as the test solution. Prepare the control solution
with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more
conditions, the relative standard deviations of the ratio of
than 0.019z).
the peak area of acetone, methanol and mesityl oxide to that
of the internal standard are not more than 4.0z, respec-
Cilazapril Hydrate according to Method 4, and perform the
tively.
test. However, use 10 mL of a solution of magnesium nitrate
Water <2.48> Not more than 2.0z (0.5 g, volumetric titra- hexahydrate in ethanol (95) (1 in 8). Prepare the control so-
tion, direct titration). lution with 2.0 mL of Standard Lead Solution (not more
than 20 ppm).
Assay Weigh accurately about 0.3 g of Cilastatin Sodium,
(4) Related substances—Dissolve 0.10 g of Cilazapril
dissolve in 30 mL of methanol, add 5 mL of water, and
Hydrate in 20 mL of methanol, and use this solution as the
adjust to pH 3.0 with 0.1 mol/L hydrochloric acid TS.
Titrate <2.50> with 0.1 mol/L sodium hydroxide VS from the
first equivalence point to the third equivalence point (poten-
the standard solution (1). Pipet 3 mL of the standard solu-
JP XVII Official Monographs / Cilazapril Tablets 703
tion (1), add methanol to make exactly 10 mL, and use this solutions are dark brown and they show the same Rf value.
solution as the standard solution (2). Separately, pipet 2 mL
of the standard solution (1), add methanol to make exactly
10 mL, and use this solution as the standard solution (3).
To 1 tablet of Cilazapril Tablets add 5 mL of a mixture of
water and acetonitrile (7:3), shake well until disintegration,
solution and three standard solutions on a plate of silica gel
add the mixture of water and acetonitrile (7:3) to make
with fluorescent indicator for thin-layer chromatography.
exactly V mL so that each mL contains about 25 mg of
Develop the plate with a mixture of ethyl acetate, methanol,
cilazapril (C22H31N3O5), and centrifuge. Pipet 4 mL of the
acetic acid (100), hexane, and water (62:15:10:10:3) to a dis-
supernatant liquid, add exactly 1 mL of the internal standard
tance of about 15 cm, and air-dry the plate. Leave the plate
solution, add the mixture of water and acetonitrile (7:3) to
in iodine vapor for 2 hours, and examine the plate under ul-
traviolet light (main wavelength: 254 nm): of the spots other
Separately, weigh accurately about 26 mg of cilazapril for
than the principal spot with an R f value close to 0.40 ob-
assay (separately determine the water <2.48> in the same
tained from the sample solution, the spot in the vicinity of
manner as Cilazapril Hydrate), and dissolve in the mixture
R f value 0.17 is not more intense than the spot obtained
of water and acetonitrile (7:3) to make exactly 50 mL. Pipet
from the standard solution (1), and the spot in the vicinity of
2 mL of this solution, add exactly 10 mL of the internal
R f value 0.44 is not more intense than the spot from the
standard solution, add the mixture of water and acetonitrile
standard solution (2). The number of all other spot does not
(7:3) to make 100 mL, and use this solution as the standard
exceed 3, and of these spots, no more than one is more in-
solution. Perform the test with 100 mL each of the sample so-
tense than the spot from the standard solution (3) and none
lution and standard solution as directed under Liquid Chro-
are more intense than the spot from the standard solution
(2).
and calculate the ratios, QT and QS, of the peak area of
Water <2.48> 3.5 – 5.0z (0.3 g, volumetric titration, direct cilazapril to that of the internal standard.
titration).
Amount (mg) of cilazapril (C22H31N3O5)
Residue on ignition <2.44> Not more than 0.1z (0.5 g). ＝ MS × QT/QS × V/1000
Assay Weigh accurately about 0.2 g of Cilazapril Hydrate, MS: Amount (mg) of cilazapril for assay taken, calculated
dissolve in 50 mL of acetic acid (100), and titrate <2.50> with on the anhydrous basis
0.02 mol/L perchloric acid VS (potentiometric titration).
Perform a blank determination in the same manner and
late in a mixture of water and acetonitrile (7:3) (1 in 12,500).
Each mL of 0.02 mol/L perchloric acid VS Proceed as directed in the operating conditions in the
＝ 8.350 mg of C22H31N3O5 Assay.
mL of the standard solution under the above conditions,
cilazapril and the internal standard are eluted in this order
with the resolution between these peaks being not less than 6.
Cilazapril Tablets System repeatability: When the test is repeated 6 times
with 100 mL of the standard solution under the above condi-
シラザプリル錠
tions, the relative standard deviation of the ratio of the peak
area of cilazapril to that of the internal standard is not more
Cilazapril Tablets contain not less than 93.0z and than 2.0z.
cilazapril (C22H31N3O5: 417.50).
Method of preparation Prepare as directed under Tablets, mL of water as the dissolution medium, the dissolution rate
with Cilazapril Hydrate. in 15 minutes of Cilazapril Tablets is not less than 85z.
Start the test with 1 tablet of Cilazapril Tablets, withdraw
Identification To a quantity of powdered Cilazapril
Tablets, equivalent to 2 mg of cilazapril (C22H31N3O5), add 2
mL of a mixture of acetonitrile and ethyl acetate (3:1),
shake, treat with ultrasonic waves for 30 seconds, centrifuge,
mL of the filtrate, pipet V mL of the subsequent filtrate, and
and use the supernatant liquid as the sample solution. Sepa-
rately, dissolve 5 mg of cilazapril in 5 mL of the mixture of
about 0.28 mg of cilazapril (C22H31N3O5). Pipet 10 mL of the
acetonitrile and ethyl acetate (3:1), and use this solution as
solution, add exactly 5 mL of acetonitrile, and use this solu-
about 29 mg of cilazapril for assay (separately determine the
water <2.48> in the same manner as Cilazapril Hydrate), and
dissolve in water to make exactly 100 mL. Pipet 5 mL of the
chromatography. Develop the plate with a mixture of ethyl
solution, add water to make exactly 100 mL. Then, pipet 2
acetate, methanol, acetic acid (100), hexane and water
mL of this solution, add water to make exactly 100 mL.
(62:15:10:10:3) to a distance of about 15 cm, and air-dry the
Pipet 10 mL of this solution, add exactly 5 mL of aceto-
plate. Place the plate in iodine vapor for 2 hours, and imme-
nitrile, and use this solution as the standard solution. Per-
diately examine under ultraviolet light (main wavelength:
254 nm): the spots obtained from the sample and standard
704 Cilnidipine / Official Monographs JP XVII
and standard solution as directed under Liquid Chromatog- late in a mixture of water and acetonitrile (7:3) (1 in 12,500).
raphy <2.01> according to the following conditions, and de- Operating conditions—
termine the peak areas, AT and AS, of cilazapril in each solu- Detector: An ultraviolet absorption photometer (wave-
tion. length: 210 nm).
of cilazapril (C22H31N3O5)
＝ MS × AT/AS × V?/V × 1/C × 9/10
MS: Amount (mg) of cilazapril for assay taken, calculated 239C.
on the anhydrous basis Mobile phase: To a solution consisting of 180 mL of tetra-
C: Labeled amount (mg) of cilazapril (C22H31N3O5) in 1 hydrofuran for liquid chromatography, 120 mL of aceto-
tablet nitrile for liquid chromatography and 3 mL of triethylamine
add water to make 1000 mL, and adjust the pH to 2.5 with
phosphoric acid.
Flow rate: Adjust so that the retention time of cilazapril is
length: 210 nm).
about 10 minutes.
ditions, cilazapril and the internal standard are eluted in this
259 C.
Mobile phase: To a solution consisting of 180 mL of tetra-
than 6.
hydrofuran for liquid chromatography, 120 mL of aceto-
nitrile for liquid chromatography and 3 mL of triethylamine
add water to make 1000 mL, and adjust the pH to 2.5 with
phosphoric acid.
the peak area of cilazapril to that of the internal standard is
Flow rate: Adjust so that the retention time of cilazapril is
not more than 1.0z.
about 10 minutes.
System suitability— Containers and storage Containers—Tight containers.
ditions, the number of theoretical plates and the symmetry Cilnidipine
factor of the peak of cilazapril are not less than 3000 and not
more than 2.0, respectively. シルニジピン
area of cilazapril is not more than 2.0z.
Assay Weigh acurately the mass of not less than 20
Cilazapril Tablets, and powder. Weigh accurately a portion
of the powder, equivalent to about 1 mg of cilazapril
(C22H31N3O5), add 30 mL of a mixture of water and aceto-
nitrile (7:3), and treat with ultrasonic waves for 5 minutes. C27H28N2O7: 492.52
Next, add exactly 5 mL of the internal standard solution, 3-(2-Methoxyethyl) 5-[(2E )-3-phenylprop-2-en-1-yl] (4RS )-
add the mixture of water and acetonitrile (7:3) to make 50 2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydropyridine-3,5-
mL, and centrifuge. Filter the supernatant liquid through a dicarboxylate
membrane filter with a pore size not exceeding 0.5 mm, and [132203-70-4]
use the filtrate as the sample solution. Separately, weigh ac-
curately about 26 mg of cilazapril for assay (separately deter- Cilnidipine, when dried, contains not less than
mine the water <2.48> in the same manner as Cilazapril Hy- 98.0z and not more than 102.0z of cilnidipine
drate), and dissolve in the mixture of water and acetonitrile (C27H28N2O7).
(7:3) to make exactly 50 mL. Pipet 2 mL of this solution,
Description Cilnidipine occurs as a faint yellow crystalline
add exactly 5 mL of the internal standard solution, add the
powder.
mixture of water and acetonitrile (7:3) to make 50 mL, and
It is freely soluble in acetonitrile, sparingly soluble in
methanol and in ethanol (99.5), and practically insoluble in
water.
A solution of Cilnidipine in acetonitrile (1 in 100) shows
QS, of the peak area of cilazapril to that of the internal
It is gradually colored to reddish yellow and decomposed
standard.
by light.
Amount (mg) of cilazapril (C22H31N3O5)
＝ MS × QT/QS × 1/25
solution of Cilnidipine in methanol (1 in 100,000) as directed
MS: Amount (mg) of cilazapril for assay taken, calculated under Ultraviolet-visible Spectrophotometry <2.24>, and
on the anhydrous basis compare the spectrum with the Reference Spectrum or the
spectrum of a solution of Cilnidipine RS prepared in the
JP XVII Official Monographs / Cilnidipine Tablets 705
similar intensities of absorption at the same wavelengths. solution and the standard solution, respectively. Perform the
(2) Determine the infrared absorption spectrum of previ- test with 10 mL each of the sample solution and standard so-
ously dried Cilnidipine as directed in the potassium bromide lution as directed under Liquid Chromatography <2.01> ac-
disk method under Infrared Spectrophotometry <2.25>, and cording to the following conditions, and calculate the ratios,
compare the spectrum with the Reference Spectrum or the QT and QS, of the peak area of cilnidipine to that of the in-
spectrum of dried Cilnidipine RS: both spectra exhibit simi- ternal standard.
Amount (mg) of cilnidipine (C27H28N2O7) ＝ MS × QT/QS
MS: Amount (mg) of Cilnidipine RS taken
Cilnidipine according to Method 4, and perform the test.
length: 240 nm).
light-resistant vessels. Dissolve 50 mg of Cilnidipine in 20
mL of acetonitrile, add the mobile phase to make 100 mL,
ter and 25 cm in length, packed with perfluorohexylpropyl-
the sample solution, add the mobile phase to make exactly
cle diameter).
259C.
Mobile phase: Dissolve 1.36 g of sodium acetate trihydrate
in water to make 1000 mL, and adjust to pH 5.5 with diluted
termine each peak area by the automatic integration method:
acetic acid (100) (1 in 100). To 400 mL of this solution add
the area of the peak, having the relative retention time of
600 mL of methanol.
about 0.5 to cilnidipine, obtained from the sample solution
Flow rate: Adjust so that the retention time of cilnidipine
is not larger than 2/5 times the peak area of cilnidipine ob-
tained from the standard solution, the area of the peaks
other than cilnidipine and the above mentioned peak from
System performance: After exposing Cilnidipine to a fluo-
the sample solution is not larger than 1/5 times the peak area
rescent light (15,000 lx･h), take 10 mg, dissolve in 4 mL of
of cilnidipine from the standard solution, and the total area
acetonitrile, and add the mobile phase to make 20 mL. When
of the peaks other than cilnidipine from the sample solution
the procedure is run with 10 mL of this solution under the
is not larger than the peak area of cilnidipine from the stand-
above operating conditions, the resolution between the peak
ard solution. For the area of the peak, having the relative
of cilnidipine and the peak having the relative retention time
retention time of about 1.15, about 1.6, and about 1.7 to cil-
of about 1.07 to cilnidipine is not less than 1.5.
nidipine, multiply the relative response factor, 1.5, 1.4, and
1.6, respectively.
the peak area of cilnidipine to that of the internal standard is
not more than 1.0z.
the Assay.
Time span of measurement: About 3 times as long as the Containers and storage Containers—Tight containers.
retention time of cilnidipine, beginning after the solvent Storage—Light-resistant.
peak.
System performance: Proceed as directed in the system Cilnidipine Tablets
Test for required detectability: Pipet 5 mL of the standard シルニジピン錠
firm that the peak area of cilnidipine obtained from 10 mL of
Cilnidipine Tablets contain not less than 95.0z and
this solution is equivalent to 7 to 13z of that obtained from
not more than 105.0z of the labeled amount of cil-
nidipine (C27H28N2O7: 492.52).
with 10 mL of the standard solution under the above operat- Method of preparation Prepare as directed under Tablets,
ing conditions, the relative standard deviation of the peak with Cilnidipine.
area of cilnidipine is not more than 2.0z.
Identification Powder Cilnidipine Tablets. To a portion of
Loss on drying <2.41> Not more than 0.5z (1.0 g, in vacu- the powder, equivalent to 20 mg of Cilnidipine, add 20 mL
um, 609C, 3 hours). of methanol, shake well, and centrifuge. To 1 mL of the su-
pernatant liquid add methanol to make 100 mL. Determine
the absorption spectrum of this solution as directed under
Assay Conduct this procedure using light-resistant vessels. Ultraviolet-visible Spectrophotometry <2.24>: it exhibits
Weigh accurately about 50 mg each of Cilnidipine and Cil- maxima between 238 nm and 242 nm and between 350 nm
nidipine RS, both previously dried, dissolve in 20 mL of and 360 nm.
acetonitrile, and add the mobile phase to make exactly 100
Purity Related substances—Conduct this procedure using
mL, respectively. Pipet 5 mL each of these solutions, add ex-
light-resistant vessels. Powder Cilnidipine Tablets. To a por-
actly 5 mL of the internal standard solution, add the mobile
tion of the powder, equivalent to 25 mg of Cilnidipine, add
phase to make 25 mL, and use these solutions as the sample
40 mL of the mobile phase, shake well, and add the mobile
706 Cilnidipine Tablets / Official Monographs JP XVII
phase to make 50 mL. Centrifuge, and use the supernatant Dissolution <6.10> When the test is performed at 75 revolu-
liquid as the sample solution. Pipet 3 mL of the sample solutions per minute according to the Paddle method, using 900
tion, add the mobile phase to make exactly 200 mL, and use mL of a solution of polysorbate 80 (dissolving 1 g of poly-
this solution as the standard solution. Perform the test with sorbate 80 in 1000 mL of 2nd fluid for dissolution test) as the
exactly 20 mL each of the sample solution and standard solu- dissolution medium, the dissolution rate in 90 minutes of
tion as directed under Liquid Chromatography <2.01> ac- Cilnidipine Tablets is not less than 70z.
cording to the following conditions, and determine each Start the test with 1 tablet of Cilnidipine Tablets, with-
peak area by the automatic integration method: the area of draw not less than 20 mL of the medium at the specified
the peak, having the relative retention time of about 1.09 to minute after starting the test, and filter through a membrane
cilnidipine, obtained from the sample solution is not larger filter with a pore size not exceeding 0.45 mm. Discard the
than 1/3 times the peak area of cilnidipine obtained from the first 10 mL of the filtrate, pipet V mL of the subsequent
standard solution, the area of the peaks other than cilnidi- filtrate, add the dissolution medium to make exactly V? mL
pine and the peak mentioned above from the sample solution so that each mL contains about 5.6 mg of cilnidipine
is not larger than 2/15 times the peak area of cilnidipine (C27H28N2O7), and use this solution as the sample solution.
from the standard solution, and the total area of the peaks Separately, weigh accurately about 28 mg of Cilnidipine RS,
other than cilnidipine from the sample solution is not larger previously dried in vacuum at 609C for 3 hours, dissolve in
than the peak area of cilnidipine from the standard solution. acetonitrile to make exactly 100 mL. Pipet 2 mL of this solu-
For the area of the peak, having the relative retention time tion, add the dissolution medium to make exactly 100 mL,
of about 1.09 to cilnidipine, multiply the relative response and use this solution as the standard solution. Perform the
factor, 1.4. test with exactly 20 mL each of the sample solution and
Operating conditions— standard solution as directed under Liquid Chromatography
Detector, column, column temperature, mobile phase, and <2.01> according to the following conditions, and determine
flow rate: Proceed as directed in the operating conditions in the peak areas, AT and AS, of cilnidipine in each solution.
the Assay.
of cilnidipine (C27H28N2O7)
retention time of cilnidipine, beginning after the solvent
＝ MS × AT/AS × V?/V × 1/C × 18
peak.
System suitability— MS: Amount (mg) of Cilnidipine RS taken
Test for required detectability: Pipet 5 mL of the standard C: Labeled amount (mg) of cilnidipine (C27H28N2O7) in 1
solution, add the mobile phase to make exactly 150 mL. tablet
Confirm that the peak area of cilnidipine obtained with 20
mL of this solution is equivalent to 2.4 to 4.3z of that ob-
Detector: An ultraviolet absorption photometer (wave
length: 240 nm).
mL of standard solution under the above operating condi-
tor of the peak of cilnidipine are not less than 15,000 and not
409C.
phate dodecahydrate in 1000 mL of water, and adjust to pH
6.0 with phosphoric acid. To 400 mL of this solution add
area of cilnidipine is not more than 2.0z.
600 mL of acetonitrile.
Uniformity of dosage units <6.02> Perform the test accord- Flow rate: Adjust so that the retention time of cilnidipine
ing to the following method: it meets the requirement of the is about 8 minutes.
Content uniformity test. System suitability—
Conduct this procedure using light-resistant vessels. To 1 System performance: When the procedure is run with 20
tablet of Cilnidipine Tablets add V/10 mL of water, and mL of the standard solution under the above operating con-
shake to completely disintegrate the tablet. Add acetonitrile ditions, the number of theoretical plates and the symmetry
to make exactly V mL so that each mL contains about 0.2 factor of the peak of cilnidipine are not less than 2000 and
mg of cilnidipine (C27H28N2O7), and centrifuge. Pipet 4 mL not more than 2.0, respectively.
of the supernatant liquid, add a mixture of acetonitrile and System repeatability: When the test is repeated 6 times
water (9:1) to make exactly 20 mL, filter, if necessary, and with 20 mL of the standard solution under the above operat-
use this solution as the sample solution. Separately, weigh ing conditions, the relative standard deviation of the peak
accurately about 20 mg of Cilnidipine RS, previously dried area of cilnidipine is not more than 2.0z.
in vacuum at 609 C for 3 hours, dissolve in a mixture of
Assay Conduct this procedure using light-resistant vessels.
acetonitrile and water (9:1) to make exactly 50 mL. Pipet 5
Weigh accurately the mass of not less than 20 Cilnidipine
mL of this solution, add a mixture of acetonitrile and water
Tablets, and powder. Weigh accurately a portion of the
powder, equivalent to about 25 mg of cilnidipine
(C27H28N2O7), add 40 mL of the mobile phase, shake well,
355 nm of the sample solution and standard solution as
and add the mobile phase to make exactly 50 mL. Centrifuge
this solution, pipet 5 mL of the supernatant liquid, add
using a mixture of acetonitrile and water (9:1) as the control.
exactly 2.5 mL of the internal standard solution, add the
Amount (mg) of cilnidipine (C27H28N2O7) mobile phase to make 25 mL, and use this solution as the
＝ MS × AT/AS × V/100 sample solution. Separately, weigh accurately about 25 mg
of Cilnidipine RS, previously dried in vacuum at 609C for 3
MS: Amount (mg) of Cilnidipine RS taken
hours, dissolve in the mobile phase to make exactly 50 mL.
JP XVII Official Monographs / Cilostazol 707
Pipet 5 mL of this solution, add exactly 2.5 mL of the inter- solution of Cilostazol in methanol (1 in 100,000) as directed
nal standard solution, add the mobile phase to make 25 mL, under Ultraviolet-visible Spectrophotometry <2.24>, and
and use this solution as the standard solution. Perform the compare the spectrum with the Reference Spectrum or the
test with 10 mL each of the sample solution and standard so- spectrum of a solution of Cilostazol RS prepared in the same
lution as directed under Liquid Chromatography <2.01> ac- manner as the sample solution: both spectra exhibit similar
cording to the following conditions, and calculate the ratios, intensities of absorption at the same wavelengths.
QT and QS, of the peak area of cilnidipine to that of the in- (2) Determine the infrared absorption spectrum of
ternal standard. Cilostazol as directed in the potassium bromide disk method
Amount (mg) of cilnidipine (C27H28N2O7) ＝ MS × QT/QS
MS: Amount (mg) of Cilnidipine RS taken Cilostazol RS: both spectra exhibit similar intensities of ab-
Internal standard solution—A solution of 4,4'-difluoroben-
zophenone in the mobile phase (1 in 500). Melting point <2.60> 158 – 1629C
Cilostazol according to Method 2, and perform the test. Pre-
length: 240 nm).
(2) Related substances—Dissolve 25 mg of Cilostazol in
25 mL of acetonitrile, and use this solution as the sample so-
lution. Pipet 1 mL of the sample solution, and add aceto-
409 C.
nitrile to make exactly 100 mL. Pipet 10 mL of this solution,
add acetonitrile to make exactly 50 mL, and use this solution
phate dodecahydrate in 1000 mL of water, and adjust to pH
6.0 with phosphoric acid. To 400 mL of this solution add
600 mL of acetonitrile.
Flow rate: Adjust so that the retention time of cilnidipine
tomatic integration method: the area of the peak other than
cilostazol obtained with the sample solution is not larger
than 7/10 times the peak area of cilostazol obtained with the
ditions, the internal standard and cilnidipine are eluted in
the peak of cilostazol with the sample solution is not larger
than 1.2 times the peak area of cilostazol with the standard
less than 15.
solution.
length: 254 nm).
the peak area of cilnidipine to that of the internal standard is
not more than 1.0z.
Containers and storage Containers—Tight containers. chromatography (5 mm in particle diameter).
Storage—Light-resistant. Column temperature: A constant temperature of about
259C.
Mobile phase: A mixture of hexane, ethyl acetate and
Cilostazol methanol (10:9:1).
Flow rate: Adjust so that the retention time of cilostazol is
シロスタゾール about 7 minutes.
retention time of cilostazol, beginning after the solvent peak.
solution, and add acetonitrile to make exactly 10 mL. Con-
firm that the peak area of cilostazol obtained with 10 mL of
this solution is equivalent to 7 to 13z of that obtained with
C20H27N5O2: 369.46
System performance: To 1 mL of the sample solution, add
6-[4-(1-Cyclohexyl-1H-tetrazol-5-yl)butyloxy]-
1 mL of a solution prepared by dissolving 5 mg of 3,4-
3,4-dihydroquinolin-2(1H )-one
dihydro-6-hydroxy-2(1H )-quinolinone in 10 mL of aceto-
[73963-72-1]
nitrile and acetonitrile to make 100 mL. When the procedure
is run with 10 mL of this solution under the above operating
Cilostazol, when dried, contains not less than 98.5z
conditions, 3,4-dihydro-6-hydroxy-2(1H)-quinolinone and
and not more than 101.5z of cilostazol (C20H27N5O2).
cilostazol are eluted in this order with the resolution between
Description Cilostazol occurs as white to pale yellowish these peaks being not less than 9.
white, crystals or crystalline powder. System repeatability: When the test is repeated 6 times
It is slightly soluble in methanol, in ethanol (99.5) and in with 10 mL of the standard solution under the above operat-
acetonitrile, and practically insoluble in water. ing conditions, the relative standard deviation of the peak
area of cilostazol is not more than 2.0z.
708 Cilostazol Tablets / Official Monographs JP XVII
Loss on drying <2.41> Not more than 0.1z (1 g, 1059C, gel for thin-layer chromatography, develop the plate with a
2 hours). mixture of ethyl acetate, acetonitrile, methanol and formic
acid (75:25:5:1) to a distance of about 12 cm, and air-dry the
plate. Spray evenly Dragendorff's TS for spraying on the
Assay Weigh accurately about 50 mg each of Cilostazol plate: the principal spot with the sample solution and the
and Cilostazol RS, previously dried, dissolve each in a suita- spot with the standard solution are orange in color and have
ble amount of methanol, add exactly 5 mL of the internal the same R f value.
standard solution and methanol to make 50 mL. To 1 mL
each of these solutions add methanol to make 10 mL, and
use these solutions as the sample solution and the standard
solution, respectively. Perform the test with 10 mL each of
To 1 tablet of Cilostazol Tablets add 2 mL of water to
disintegrate the tablet, add the internal standard solution
exactly 5 mL for a 50-mg tablet and exactly 10 mL for a
100-mg tablet, and add methanol to make 50 mL. Shake for
area of cilostazol to that of the internal standard.
10 minutes for the 50-mg tablet and for 20 minutes for the
Amount (mg) of cilostazol (C20H27N5O2) ＝ MS × QT/QS 100-mg tablet. To 1 mL of the solution add methanol to
make 10 mL for the 50-mg tablet and 20 mL for the 100-mg
MS: Amount (mg) of Cilostazol RS taken
tablet, filter through a membrane filter with a pore size not
Internal standard solution—A solution of benzophenone in exceeding 0.5 mm, and use the filtrate as the sample solution.
methanol (1 in 250). Proceed as directed in the Assay.
Amount (mg) of cilostazol (C20H27N5O2)
＝ MS × QT/QS × C/50
length: 254 nm).
Column: A stainless steel column 4.6 mm in inside diame- MS: Amount (mg) of Cilostazol RS taken
ter and 15 cm in length, packed with octadecylsilanized silica C: Labeled amount (mg) of cilostazol (C20H27N5O2) in 1
gel for liquid chromatography (5 mm in particle diameter). tablet
259 C.
methanol (1 in 250).
Mobile phase: A mixture of water, acetonitrile and metha-
nol (10:7:3). Dissolution <6.10> When the test is performed at 50 revolu-
Flow rate: Adjust so that the retention time of cilostazol is tions per minute according to the Paddle method, using 900
about 9 minutes. mL of a solution of sodium lauryl sulfate (3 in 1000) as the
System suitability— dissolution medium, the dissolution rates of a 50-mg tablet
System performance: When the procedure is run with 10 in 45 minutes and a 100-mg tablet in 60 minutes are not less
mL of the standard solution under the above operating con- than 75z and not less than 70z, respectively.
ditions, cilostazol and the internal standard are eluted in this Start the test with 1 tablet of Cilostazol Tablets, withdraw
order with the resolution between these peaks being not less not less than 20 mL of the medium at the specified minute
than 9. after starting the test, and filter through a membrane filter
System repeatability: When the test is repeated 5 times with a pore size not exceeding 0.45 mm. Discard the first 10
with 10 mL of the standard solution under the above operat- mL of the filtrate, pipet V mL of the subsequent filtrate, add
ing conditions, the relative standard deviation of the ratio of the dissolution medium to make exactly V? mL so that each
the peak area of cilostazol to that of the internal standard is mL contains about 5.6 mg of cilostazol (C20H27N5O2), and
not more than 1.0z. use this solution as the sample solution. Separately, weigh
accurately about 28 mg of Cilostazol RS, previously dried at
1059C for 2 hours, and dissolve in methanol to make exactly
ers.
100 mL. Pipet 4 mL of this solution, add the dissolution
Cilostazol Tablets of the sample solution and standard solution at 257 nm as
シロスタゾール錠
using the dissolution medium as the control.
Cilostazol Tablets contain not less than 95.0z and
of cilostazol (C20H27N5O2)
not more than 105.0z of the labeled amount of ＝ MS × AT/AS × V?/V × 1/C × 18
cilostazol (C20H27N5O2: 369.46).
MS: Amount (mg) of Cilostazol RS taken
C: Labeled amount (mg) of cilostazol (C20H27N5O2) in 1
with Cilostazol.
tablet
Identification Mix well an amount of powdered Cilostazol
Tablets, equivalent to 50 mg of Cilostazol, with 10 mL of
Cilostazol Tablets, and powder. Weigh accurately a portion
acetone, centrifuge, and use the supernatant liquid as the
of the powder, equivalent to about 50 mg of cilostazol
sample solution. Separately, dissolve 25 mg of Cilostazol RS
(C20H27N5O2), add exactly 5 mL of the internal standard
in 5 mL of acetone, and use this solution as the standard so-
solution and methanol to make 50 mL, and shake well for
lution. Perform the test with these solutions as directed un-
10 minutes. To 1 mL of this solution add methanol to make
der Thin-layer Chromatography <2.03>. Spot 6 mL each of
10 mL, filter through a membrane filter with a pore size not
exceeding 0.5 mm, and use the filtrate as the sample solution.
JP XVII Official Monographs / Cinoxacin 709
Separately, weigh accurately about 50 mg of Cilostazol RS, boiled and cooled water, shake for 5 minutes and filter: the
previously dried at 1059 C for 2 hours, dissolve in a suitable pH of the filtrate is between 9.0 and 10.5.
amount of methanol, and add exactly 5 mL of the internal
standard solution, and add methanol to make 50 mL. To 1
mL of this solution add methanol to make 10 mL, and use Purity (1) Clarity and color of solution—Dissolve 1.0 g
this solution as the standard solution. Perform the test with of Cimetidine in 10 mL of methanol: the solution is clear
10 mL each of the sample solution and standard solution as and colorless to pale yellow in color.
directed under Liquid Chromatography <2.01> according to (2) Heavy metals <1.07>—Proceed with 2.0 g of Cimeti-
the following conditions, and calculate the ratios, QT and dine according to Method 2, and perform the test. Prepare
QS, of the peak area of cilostazol to that of the internal the control solution with 2.0 mL of Standard Lead Solution
standard. (not more than 10 ppm).
(3) Arsenic <1.11>—Dissolve 1.0 g of Cimetidine in 5 mL
Amount (mg) of cilostazol (C20H27N5O2) ＝ MS × QT/QS
of dilute hydrochloric acid, and perform the test with this so-
MS: Amount (mg) of Cilostazol RS taken lution (not more than 2 ppm).
(4) Related substances—Dissolve 0.5 g of Cimetidine in
10 mL of methanol, and use this solution as the sample solu-
methanol (1 in 250).
tion. Pipet 1 mL of the sample solution, add methanol to
make exactly 100 mL. Pipet 1 mL of this solution, add meth-
anol to make exactly 10 mL, and use this solution as the
Assay under Cilostazol.
suitability in the Assay under Cilostazol.
the plate with a mixture of ethyl acetate, methanol and am-
monia solution (28) (21:2:2) to a distance of about 15 cm,
air-dry the plate, and then dry at 809C for 30 minutes. Allow
the peak area of cilostazol to that of the internal standard is
the plate to stand in iodine vapor for 45 minutes: the spots
not more than 1.5z.
Containers and storage Containers—Well-closed contain- not more intense than the spot from the standard solution.
ers.
3 hours).
Cimetidine Residue on ignition <2.44> Not more than 0.2z (1 g).

Assay Weigh accurately about 0.24 g of Cimetidine, previ-
シメチジン
C10H16N6S: 252.34 ＝ 25.23 mg of C10H16N6S
2-Cyano-1-methyl-3-{2-[(5-methyl-1H-imidazol-4-
yl)methylsulfanyl]ethyl}guanidine
ers.
[51481-61-9]
Cimetidine, when dried, contains not less than
99.0z of cimetidine (C10H16N6S).
Cinoxacin
Description Cimetidine occurs as a white crystalline pow-
der. It is odorless, and has a bitter taste. シノキサシン
It is freely soluble in methanol and in acetic acid (100),
sparingly soluble in ethanol (95), slightly soluble in water,
Identification (1) To 0.1 mL of a solution of Cimetidine
in ethanol (95) (1 in 100) add 5 mL of citric acid-acetic anhy- C12H10N2O5: 262.22
dride TS, and heat in a water bath for 15 minutes: a red- 5-Ethyl-8-oxo-5,8-dihydro[1,3]dioxolo[4,5-g ]cinnoline-
purple color develops. 7-carboxylic acid
(2) Determine the infrared absorption spectrum of [28657-80-9]
Cimetidine, previously dried, as directed in the potassium
bromide disk method under the Infrared Spectrophotometry Cinoxacin, when dried, contains not less than
<2.25>, and compare the spectrum with the Reference Spec- 98.0z and not more than 101.0z of cinoxacin
trum: both spectra exhibit similar intensities of absorption at (C12H10N2O5).
Description Cinoxacin occurs as a white to pale yellow
pH <2.54> Dissolve 0.5 g of Cimetidine in 50 mL of freshly crystalline powder. It is odorless or has a slight, characteris-
710 Cinoxacin Capsules / Official Monographs JP XVII
tic odor. It has a bitter taste.
It is slightly soluble in N, N-dimethylformamide and in Cinoxacin Capsules
acetone, very slightly soluble in ethanol (99.5), and practi-
cally insoluble in water. シノキサシンカプセル
Cinoxacin Capsules contain not less than 95.0z and
Identification (1) Dissolve 30 mg of Cinoxacin in 10 mL not more than 105.0z of the labeled amount of cinox-
of dilute sodium hydroxide TS, and add water to make 100 acin (C12H10N2O5: 262.22).
mL. To 1 mL of this solution add 0.1 mol/L hydrochloric
acid TS to make 50 mL. Determine the absorption spectrum
sules, with Cinoxacin.
of this solution as directed under Ultraviolet-visible Spectro-
photometry <2.24>, and compare the spectrum with the Ref- Identification To a quantity of the contents of Cinoxacin
erence Spectrum: both spectra exhibit similar intensities of Capsules, equivalent to 10 mg of Cinoxacin, add 20 mL of
absorption at the same wavelengths. acetone, shake well, and centrifuge. To 3 mL of the superna-
(2) Determine the infrared absorption spectrum of tant liquid add acetone to make 10 mL, and use this solution
Cinoxacin as directed in the potassium bromide disk method as the sample solution. Separately, dissolve 10 mg of cinoxa-
under Infrared Spectrophotometry <2.25>, and compare the cin for assay in 20 mL of acetone. To 3 mL of this solution
spectrum with the Reference Spectrum: both spectra exhibit add acetone to make 10 mL, and use this solution as the
similar intensities of absorption at the same wave numbers. standard solution. Perform the test with these solutions as
Purity (1) Sulfate <1.14>—Dissolve 0.20 g of Cinoxacin in
10 mL of dilute sodium hydroxide TS, add 20 mL of 0.1
mol/L hydrochloric acid TS, shake, filter, and add water to
chromatography. Develop the plate with a mixture of aceto-
the filtrate to make 50 mL. Perform the test using this solu-
nitrile, water and ammonia solution (28) (14:4:1) to a dis-
0.20 mL of 0.005 mol/L sulfuric acid VS by adding 10 mL of
ultraviolet light (main wavelength: 254 nm): the principal
dilute sodium hydroxide TS, 20 mL of 0.1 mol/L hydrochlo-
spot obtained from the sample solution and the spot ob-
ric acid TS, and water to make 50 mL (not more than
tained from the standard solution show a blue-purple color
0.048z).
and the same R f value.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Cinoxa-
cin according to Method 2, and perform the test. Prepare the Uniformity of dosage units <6.02> Perform the test accord-
control solution with 2.0 mL of Standard Lead Solution (not ing to the following method: it meets the requirement of the
more than 20 ppm). Content uniformity test.
(3) Related substances—Dissolve 10 mg of Cinoxacin in To 1 capsule of Cinoxacin Capsules add 40 mL of dilute
10 mL of acetone, and use this solution as the sample solu- sodium hydroxide TS, and dissolve the capsule in lukewarm
tion. Pipet 1 mL of the sample solution, add acetone to water with occasional shaking. After cooling, add water and
make exactly 200 mL, and use this solution as the standard shake well, add water to make exactly V mL so that each mL
solution. Perform the test with these solutions as directed contains about 1 mg of cinoxacin (C12H10N2O5), and filter.
under Thin-layer Chromatography <2.03>. Spot 10 mL each Discard the first 20 mL of the filtrate, pipet 1 mL of the sub-
of the sample solution and standard solution on a plate of sequent filtrate, add 0.1 mol/L hydrochloric acid TS to
silica gel with fluorescent indicator for thin-layer chromatog- make exactly 100 mL, and use this solution as the sample so-
raphy. Develop the plate with a mixture of acetonitrile, lution. Separately, weigh accurately about 0.2 g of cinoxacin
water and ammonia solution (28) (14:4:1) to a distance of for assay, previously dried at 1059C for 1 hour, dissolve in
about 10 cm, and air-dry the plate. Examine under ultravio- 40 mL of dilute sodium hydroxide TS, and add water to
let light (main wavelength: 254 nm): the spots other than the make exactly 200 mL. Pipet 1 mL of this solution, add 0.1
principal spot obtained from the sample solution are not mol/L of hydrochloric acid TS to make exactly 100 mL, and
more intense than the spot obtained from the standard solu- use this solution as the standard solution. Perform the test
tion. with the sample solution and standard solution as directed
under Ultraviolet-visible Spectrophotometry <2.24>, and de-
termine the absorbances, AT and AS, at 354 nm.
1 hour).
Amount (mg) of cinoxacin (C12H10N2O5)
＝ MS × AT/AS × V/200
Assay Weigh accurately about 0.4 g of Cinoxacin, previ-
MS: Amount (mg) of cinoxacin for assay taken
ously dried, add 60 mL of a mixture of acetic anhydride and
acetic acid (100) (7:3), and dissolve by warming. After cool- Dissolution <6.10> When the test is performed at 50 revolu-
ing, titrate <2.50> with 0.1 mol/L perchloric acid VS (poten- tions per minute according to the Paddle method using the
tiometric titration). Perform a blank determination in the sinker, using 900 mL of 2nd solution for dissolution test as
same manner, and make any necessary correction. the dissolution medium, the dissolution rate in 90 minutes of
Cinoxacin Capsules is not less than 70z.
Start the test with 1 capsule of Cinoxacin Capsules, with-
＝ 26.22 mg of C12H10N2O5
Containers and storage Containers—Tight containers. minute after starting the test, and filter through a membrane
filtrate, add the dissolution medium to make exactly V? mL
so that each mL contains about 11 mg of cinoxacin
JP XVII Official Monographs / Ciprofloxacin 711
(C12H10N2O5), and use this solution as the sample solution. ish white, crystalline powder.
Separately, weigh accurately about 22 mg of cinoxacin for It is practically insoluble in water and in ethanol (99.5).
assay, previously dried at 1059C for 1 hour, and dissolve in It dissolves in ammonia TS.
the dissolution medium to make exactly 100 mL. Pipet 5 mL It is gradually colored to yellow tint by light.
of this solution, add the dissolution medium to make exactly Melting point: about 2709C (with decomposition).
form the test with the sample solution and standard solution
trum of Ciprofloxacin, as directed in the potassium bromide
<2.24>, and determine the absorbances, AT and AS, at 351
nm.
spectrum of the Ciprofloxacin RS: both spectra exhibit simi-
Dissolution rate (z) with respect to the labeled amount lar intensities of absorption at the same wave numbers.
of cinoxacin (C12H10N2O5) (2) Conduct this procedure using light-resistant vessels.
＝ MS × AT/AS × V?/V × 1/C × 45 Dissolve 50 mg each of Ciprofloxacin and Ciprofloxacin RS
in 5 mL of ammonia TS, and use these solutions as the sam-
ple solution and the standard solution, respectively. Perform
C: Labeled amount (mg) of cinoxacin (C12H10N2O5) in 1
capsule
Assay Weigh accurately the mass of not less than 20 tion and the standard solution on a plate of silica gel with
Cinoxacin Capsules, take out the contents, and powder. fluorescent indicator for thin-layer chromatography. After
Wash the capsule shells with a small amount of diethyl ether, allowing to stand this plate in the vapor of ammonia for 15
allow to stand at room temperature to vaporize the diethyl minutes, develop the plate with a mixture of methanol,
ether, weigh accurately the mass of the capsule shells, and dichloromethane, ammonia solution (28) and acetonitrile
calculate the mass of the contents. Weigh accurately a por- (4:4:2:1) to a distance of about 10 cm, and air-dry the plate.
tion of the powder, equivalent to about 50 mg of cinoxacin Examine under ultraviolet light (main wavelength: 254 nm):
(C12H10N2O5), add 10 mL of dilute sodium hydroxide TS, the principal spot obtained from the sample solution and the
shake, add water to make exactly 100 mL, and filter. Discard spot obtained from the standard solution show the same Rf
the first 20 mL of the filtrate, pipet 1 mL of the subsequent value.
filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly
Purity (1) Chloride <1.03>—To 1.5 g of Ciprofloxacin
add 75 mL of water, and boil for 5 minutes. After cooling,
rately, weigh accurately about 50 mg of cinoxacin for assay,
add water to make 75 mL, and filter. To 25 mL of the fil-
previously dried at 1059C for 1 hour, dissolve in 10 mL of
trate add 6 mL of dilute nitric acid and water to make 50
dilute sodium hydroxide TS, and add water to make exactly
mL. Perform the test using this solution as the test solution.
Prepare the control solution as follows: to 0.30 mL of 0.01
chloric acid TS to make exactly 50 mL, and use this solution
mol/L hydrochloric acid VS add 6 mL of dilute sulfuric acid
as the standard solution. Perform the test with the sample
solution and standard solution as directed under Ultraviolet-
visible Spectrophotometry <2.24>, and determine the absor-
Ciprofloxacin according to Method 4, and perform the test.
bances, AT and AS, at 354 nm.
Amount (mg) of cinoxacin (C12H10N2O5) Solution (not more than 10 ppm).
＝ M S × AT / AS (3) Fluoroquinolonic acid—Conduct this procedure
using light-resistant vessels. Dissolve 50 mg of Ciprofloxacin
in ammonia TS to make exactly 5 mL, and use this solution
Containers and storage Containers—Well-closed contain- as the sample solution. Separately, dissolve 10 mg of fluoro-
ers. quinolonic acid for thin-layer chromatography in 0.1 mL of
ammonia TS and water to make exactly 100 mL. Pipet 2 mL
of this solution, add water to make exactly 10 mL, and use
Ciprofloxacin this solution as the standard solution. Perform the test with
シプロフロキサシン phy <2.03>. Spot 5 mL each of the sample solution and the
indicator for thin-layer chromatography. After allowing to
stand this plate in the vapor of ammonia for 15 minutes, de-
velop the plate with a mixture of methanol, dichlorometh-
ane, ammonia solution (28) and acetonitrile (4:4:2:1) to a
distance of about 10 cm, and air-dry the plate. Examine
under ultraviolet light (main wavelength: 254 nm): the spot
C17H18FN3O3: 331.34 obtained from the sample solution, corresponding to the
1-Cyclopropyl-6-fluoro-4-oxo-7-(piperazin-1-yl)-1,4- spot obtained from the standard solution, is not more in-
dihydroquinoline-3-carboxylic acid tense than that obtained from the standard solution.
[85721-33-1] (4) Related substances—Conduct this procedure using
light-resistant vessels. To 25 mg of Ciprofloxacin add 2 mL
Ciprofloxacin, when dried, contains not less than of a mixture of water and phosphoric acid (13:1), then add
98.5z and not more than 101.0z of ciprofloxacin the mobile phase to make 50 mL, and use this solution as the
(C17H18FN3O3). sample solution. Pipet 2 mL of the sample solution, add the
mobile phase to make exactly 20 mL. Pipet 1 mL of this so-
Description Ciprofloxacin occurs as a white to light yellow-
lution, add the mobile phase to make exactly 50 mL, and use
712 Ciprofloxacin Hydrochloride Hydrate / Official Monographs JP XVII
this solution as the standard solution. Perform the test with Flow rate: Adjust so that the retention time of ciprofloxa-
exactly 50 mL each of the sample solution and standard solu- cin is about 7 minutes.
tion as directed under Liquid Chromatography <2.01> ac- System suitability—
cording to the following conditions. Determine each peak System performance: When the procedure is run with 10
area by the automatic integration method: the area of the mL of the standard solution under the above operating con-
peak other than ciprofloxacin obtained from the sample so- ditions, the number of theoretical plates and the symmetry
lution is not larger than the peak area of ciprofloxacin ob- factor of the peak of ciprofloxacin are not less than 3500 and
tained from the standard solution, and the total area of the not more than 2.0, respectively.
peaks other than ciprofloxacin from the sample solution is System repeatability: When the test is repeated 6 times
not larger than 2.5 times the peak area of ciprofloxacin from with 10 mL of the standard solution under the above operat-
the standard solution. For the area of peak, having the rela- ing conditions, the relative standard deviation of the peak
tive retention time of about 0.4, about 0.5, and about 1.2 to area of ciprofloxacin is not more than 1.0z.
ciprofloxacin, multiply the relative response factor, 6.7, 1.3,
and 1.4, respectively.
the Assay. Ciprofloxacin Hydrochloride
Time span of measurement: About 2.3 times as long as the Hydrate
retention time of ciprofloxacin, beginning after the solvent
peak. シプロフロキサシン塩酸塩水和物
Confirm that the peak area of ciprofloxacin obtained with
mL of the standard solution under the above operating con- C17H18FN3O3.HCl.xH2O
ditions, the number of theoretical plates and the symmetry 1-Cyclopropyl-6-fluoro-4-oxo-7-(piperazin-1-yl)-1,4-
factor of the peak of ciprofloxacin are not less than 3500 and dihydroquinoline-3-carboxylic acid monohydrochloride hydrate
not more than 1.5, respectively. [86393-32-0, monohydrochloride monohydrate]
with 50 mL of the standard solution under the above operat- Ciprofloxacin Hydrochloride Hydrate contains not
ing conditions, the relative standard deviation of the peak less than 98.0z and not more than 102.0z of
area of ciprofloxacin is not more than 2.0z. ciprofloxacin hydrochloride (C17H18FN3O3.HCl:
um, 1209C, 6 hours). Description Ciprofloxacin Hydrochloride Hydrate occurs
as a white to pale yellow crystalline powder.
Assay Conduct this procedure using light-resistant vessels. nol, and very slightly soluble in ethanol (99.5).
Weigh accurately about 25 mg each of Ciprofloxacin and It is gradually colored to a slightly brownish light yellow
Ciprofloxacin RS, both dried previously, add 2 mL of a mix- by light.
ture of water and phosphoric acid (13:1), add the mobile
phase to make exactly 50 mL, and use these solutions as the
trum of Ciprofloxacin Hydrochloride Hydrate, as directed in
sample solution and the standard solution, respectively. Per-
termine the peak areas, AT and AS, of ciprofloxacin in each
(2) Conduct this procedure using light-resistant vessels.
solution.
Dissolve 50 mg of Ciprofloxacin Hydrochloride Hydrate in 5
Amount (mg) of ciprofloxacin (C17H18FN3O3) mL of water, and use this solution as the sample solution.
＝ M S × AT / AS Separately, dissolve 45 mg of Ciprofloxacin RS in 5 mL of
ammonia TS, and use this solution as the standard solution.
MS: Amount (mg) of Ciprofloxacin RS taken
Perform the test with these solutions, as directed under
Operating conditions— Thin-layer Chromatography <2.03>. Spot 5 mL each of the
Detector: An ultraviolet absorption photometer (wave- sample solution and standard solution on a plate of silica gel
length: 278 nm). with fluorescent indicator for thin-layer chromatography.
Column: A stainless steel column 4 mm in inside diameter After allowing to stand the plate in the vapor of ammonia
and 25 cm in length, packed with octadecylsilanized silica gel for 15 minutes, develop the plate with a mixture of metha-
for liquid chromatography (5 mm in particle diameter). nol, dichloromethane, ammonia solution (28) and aceto-
Column temperature: A constant temperature of about nitrile (4:4:2:1) to a distance of about 10 cm, and air-dry the
409 C. plate. Examine under ultraviolet light (main wavelength: 254
Mobile phase: To 2.88 g of phosphoric acid add water to nm): the principal spot obtained from the sample solution
make 1000 mL, and adjust to pH 3.0 with triethylamine. To and the spot obtained from the standard solution show the
870 mL of this solution add 130 mL of acetonitrile. same Rf value.
JP XVII Official Monographs / Ciprofloxacin Hydrochloride Hydrate 713
(3) A solution of Ciprofloxacin Hydrochloride Hydrate ditions, the number of theoretical plates and the symmetry
(1 in 500) responds to the Qualitative Tests <1.09> for chlo- factor of the peak of ciprofloxacin are not less than 3500 and
ride. not more than 1.5, respectively.
Purity (1) Sulfate <1.14>—Perform the test with 0.5 g of
Ciprofloxacin Hydrochloride Hydrate. Prepare the control
solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not
area of ciprofloxacin is not more than 2.0z.
more than 0.048z).
(2) Heavy metals <1.07>—Proceed with 1.0 g of Water <2.48> 4.7 – 6.7z (0.2 g, volumetric titration, direct
Ciprofloxacin Hydrochloride Hydrate according to Method titration).
2, and perform the test. Prepare the control solution with 1.0
ml of Standard Lead Solution (not more than 10 ppm).
(3) Fluoroquinolonic acid—Conduct this procedure Assay Conduct this procedure using light-resistant vessels.
using light-resistant vessels. Dissolve 50 mg of Ciprofloxacin Weigh accurately about 25 mg of Ciprofloxacin Hydrochlo-
Hydrochloride Hydrate in water to make exactly 5 mL, and ride Hydrate, dissolve in the mobile phase to make exactly 50
use this solution as the sample solution. Separately, dissolve mL, and use this solution as the sample solution. Separately,
10 mg of fluoroquinolonic acid for thin-layer chromatogra- weigh accurately about 22.5 mg of Ciprofloxacin RS, previ-
phy in 0.1 mL of ammonia TS and water to make exactly 100 ously dried at 1209C in vacuum for 6 hours, add 2 mL of a
mL. Pipet 2 mL of this solution, add water to make exactly mixture of water and phosphoric acid (13:1), then add the
10 mL, and use this solution as the standard solution. Per- mobile phase to make exactly 50 mL, and use this solution as
form the test with these solutions as directed under Thin- the standard solution. Perform the test with exactly 10 mL
layer Chromatography <2.03>. Spot 5 mL each of the sample each of the sample solution and standard solution as directed
solution and standard solution on a plate of silica gel with under Liquid Chromatography <2.01> according to the fol-
fluorescent indicator for thin-layer chromatography. After lowing operating conditions, and determine the peak areas,
allowing to stand the plate in the vapor of ammonia for 15 AT and AS, of ciprofloxacin in each solution.
minutes, develop the plate with a mixture of methanol,
Amount (mg) of ciprofloxacin hydrochloride
dichloromethane, ammonia solution (28) and acetonitrile
(C17H18FN3O3.HCl)＝ MS × AT/AS × 1.110
(4:4:2:1) to a distance of about 10 cm, and air-dry the plate.
Examine under ultraviolet light (main wavelength: 254 nm): MS: Amount (mg) of Ciprofloxacin RS taken
the spot obtained from the sample solution, corresponding
to the spot obtained from the standard solution, is not more
intense than that from the standard solution.
length: 278 nm).
light-resistant vessels. Dissolve 25 mg of Ciprofloxacin Hy-
drochloride Hydrate in 50 mL of mobile phase, and use this
lution, add the mobile phase to make exactly 20 mL. Pipet 1
409C.
mL of this solution, add the mobile phase to make exactly 50
Mobile phase: To 2.88 g of phosphoric acid add water to
make 1000 mL, and adjust to pH 3.0 with triethylamine. To
870 mL of this solution add 130 mL of acetonitrile for liquid
chromatography.
Flow rate: Adjust so that the retention time of ciprofloxa-
the peaks other than ciprofloxacin obtained from the sample
solution is not larger than the peak area of ciprofloxacin ob-
tained from the standard solution, and the total area of the
peaks other than ciprofloxacin from the sample solution is
not larger than 2.5 times the peak area of ciprofloxacin from
factor of the peak of ciprofloxacin are not less than 3500 and
the standard solution. For the area of the peaks, having the
relative retention times of about 0.4, about 0.5, and about
1.2 to ciprofloxacin, multiply the relative response factors,
6.7, 1.3, and 1.4, respectively.
area of ciprofloxacin is not more than 1.0z.
flow rate: Proceed as directed in the operating conditions in Containers and storage Containers—Tight containers.
the Assay. Storage—Light-resistant.
retention time of ciprofloxacin, beginning after the solvent
peak.
Confirm that the peak area of ciprofloxacin obtained with
714 Cisplatin / Official Monographs JP XVII
amminetrichloroplatinate is about 8 minutes.
Cisplatin System suitability—
シスプラチン mL of the standard solution under the above operating con-
factor of the peak of ammonium amminetrichloroplatinate
are not less than 1500 and not more than 2.0, respectively.
Cl2H6N2Pt: 300.05
(SP-4-2)-Diamminedichloroplatinum
area of ammonium amminetrichloroplatinate is not more
[15663-27-1]
than 3.0z.
Cisplatin, when dried, contains not less than 98.0z Loss on drying <2.41> Not more than 0.1z (1 g, 1059C,
and not more than 102.0z of cisplatin (Cl2H6N2Pt). 4 hours).
Description Cisplatin occurs as a yellow crystalline pow- Assay Conduct this procedure using light-resistant vessels.
der. Weigh accurately about 25 mg each of Cisplatin and
It is sparingly soluble in N, N-dimethylformamide, slightly Cisplatin RS, previously dried, dissolve in N, N-dimethylfor-
soluble in water, and practically insoluble in ethanol (99.5). mamide to make exactly 25 mL, and use these solutions as
the sample solution and standard solution. Perform the test
Identification (1) To 5 mL of a solution of Cisplatin (1 in
2000) add 2 to 3 drops of a solution of tin (II) chloride dihy-
drate (1 in 100): a brown precipitate is formed.
peak areas, AT and AS, of cisplatin in each solution.
Cisplatin in a solution of sodium chloride in 0.01 mol/L hy-
drochloric acid TS (9 in 1000) (1 in 2000) as directed under Amount (mg) of cisplatin (Cl2H6N2Pt) ＝ MS × AT/AS
MS: Amount (mg) of Cisplatin RS taken
of a solution of Cisplatin RS prepared in the same manner as Operating conditions—
the sample solution: both spectra exhibit similar intensities Detector: An ultraviolet absorption photometer (wave-
of absorption at the same wavelengths. length: 310 nm).
(3) Determine the infrared absorption spectrum of Column: A stainless steel column 4.6 mm in inside diame-
Cisplatin as directed in the potassium bromide disk method ter and 25 cm in length, packed with aminopropylsilanized
under Infrared Spectrophotometry <2.25>, and compare the silica gel for liquid chromatography (5 mm in particle diame-
spectrum with the Reference Spectrum or the spectrum of ter).
Cisplatin RS: both spectra exhibit similar intensities of ab- Column temperature: A constant temperature of about
sorption at the same wave numbers. 259C.
(4) A solution of Cisplatin (1 in 2000) responds to the Mobile phase: A mixture of ethyl acetate, methanol, water
Qualitative Tests <1.09> (1) for chloride. and N, N-dimethylformamide (25:16:5:5).
Flow rate: Adjust so that the retention time of cisplatin is
Purity Ammonium amminetrichloroplatinate—Conduct
about 4 minutes.
this procedure using light-resistant vessels. Dissolve 50 mg
of Cisplatin in a solution of sodium chloride (9 in 1000) to
make exactly 100 mL, and use this solution as the sample
solution. Separately, dissolve 10 mg of ammonium am-
minetrichloroplatinate for liquid chromatography, previ-
factor of the peak of cisplatin are not less than 3000 and not
ously dried at 809 C for 3 hours, in the solution of sodium
chloride (9 in 1000) to make exactly 200 mL. Pipet 2 mL of
this solution, add the solution of sodium chloride (9 in 1000)
area of cisplatin is not more than 1.0z.
uid Chromatography <2.01> according to the following con- Containers and storage Containers—Tight containers.
ditions, and determine the peak area of ammonium am-
minetrichloroplatinate by the automatic integration method:
the peak area from the sample solution is not larger than
that from the standard solution.
length: 209 nm).
chromatography having quaternary ammonium groups (10
mm in particle diameter).
259 C.
Mobile phase: A solution of ammonium sulfate (1 in 800).
Flow rate: Adjust so that the retention time of ammonium
JP XVII Official Monographs / Citicoline 715
using the solution, obtained by proceeding with 10 mL of

Citicoline water in the same manner as the sample solution, as the
blank. The amount of free phosphoric acid is not more than
シチコリン 0.1z.
Amount (z) of free phosphoric acid (H3PO4)
＝ 1/M × AT/AS × 10.32
M: Amount (mg) of Citicoline taken, calculated on the
dried basis
(5) Related substances—Dissolve 0.10 g of Citicoline in
solution. Pipet 1 mL of the sample solution, add water to
C14H26N4O11P2: 488.32
P?-[2-(Trimethylammonio)ethyl] cytidine
5?-(monohydrogen diphosphate)
[987-78-0]
tegration method: the area of the peaks other than citicoline
Citicoline contains not less than 98.0z and not
obtained from the sample solution is not larger than 3/5
more than 102.0z of citicoline (C14H26N4O11P2), cal-
times the peak area of citicolins obtained from the standard
solution, and the total area of the peaks other than citicoline
Description Citicoline occurs as a white crystalline powder. from the sample solution is not larger than the peak area of
It is very soluble in water, and practically insoluble in citicoline from the standard solution. For the area of the
ethanol (99.5). peaks, having the relative retention times of about 0.62,
It dissolves in 0.01 mol/L hydrochloric acid TS. about 0.64 and about 1.3 to citicoline, multiply the relative
response factors, 1.2, 0.7 and 0.5, respectively.
solution of Citicoline in 0.01 mol/L hydrochloric acid TS (3
in 200,000) as directed under Ultraviolet-visible Spectropho-
the Assay.
ence Spectrum or the spectrum of a solution of Citicoline RS
retention time of citicoline.
wavelengths.
Citicoline as directed in the potassium bromide disk method
that the peak area of citicoline obtained with 10 mL of this
solution is equivalent to 5.6 to 10.4z of that obtained with
Citicoline RS: both spectra exhibit similar intensities of ab-
pH <2.54> The pH of a solution obtained by dissolving ditions, the number of theoretical plates and the symmetry
1.0 g of Citicoline in 100 mL of water is between 2.5 and 3.5. factor of the peak of citicoline are not less than 2000 and 0.9
to 1.6, respectively.
of Citicoline in 8 mL of water: the solution is clear and col-
orless.
(2) Heavy metals <1.07>—Proceed with 2.0 g of Citico-
area of citicoline is not more than 2.0z.
line according to Method 1, and perform the test. Prepare
the control solution with 2.0 mL of Standard Lead Solution Loss on drying <2.41> Not more than 5.0z (1 g, in vacu-
(not more than 10 ppm). um, phosphorus (V) oxide, 1009C, 4 hours).
Assay Weigh accurately about 0.1 g of Citicoline, and dis-
of Citicoline according to Method 4, and perform the test
(4) Free phosphoric acid—Weigh accurately about 0.1 g
of Citicoline, dissolve in water to make exactly 10 mL, and
about 25 mg of Citicoline RS (separately determine the loss
use this solution as the sample solution. Separately, pipet 4
on drying <2.41> under the same conditions as Citicoline),
mL of Standard Phosphoric Acid Solution, add water to
and dissolve in water to make exactly 25 mL. Pipet 1 mL of
this solution, add water to make exactly 100 mL, and use
solution. To each of the sample solution and the standard
solution, add exactly 1 mL of hexaammonium heptamolyb-
date-sulfuric acid TS and exactly 0.5 mL of 1-amino-2-
naphthol-4-sulfonic acid TS, and after shaking, allow to
cording to the following conditions, and determine the peak
stand for 30 minutes at 20 ± 19C. To exactly 2 mL each of
areas, AT and AS, of citicoline in each solution.
these solutions add water to make exactly 10 mL, and deter-
mine the absorbances, AT and AS, of the solutions obtained Amount (mg) of citicoline (C14H26N4O11P2)
from the sample solution and the standard solution at 730 ＝ M S × AT / AS × 4
nm as directed under Ultraviolet-visible Spectrometry <2.24>,
716 Anhydrous Citric Acid / Official Monographs JP XVII
MS: Amount (mg) of Citicoline RS taken, calculated on CS, 6.0 mL of Iron (III) Chloride CS and 1.0 mL of Copper
the dried basis (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10)
to make 1000 mL.
Control solution (3): To 0.15 mL of Cobalt (II) Chloride
CS, 7.2 mL of Iron (III) Chloride CS and 0.15 mL of Cop-
length: 254 nm).
per (II) Sulfate CS add diluted dilute hydrochloric acid (1 in
Column: Combine 2 stainless steel columns (4 mm in in-
10) to make 1000 mL.
side diameter and 25 cm in length) packed with strongly bas-
(2) Sulfates—Dissolve 2.0 g of Anhydrous Citric Acid in
ic ion exchange resin for liquid chromatography (10 mm in
water to make 30 mL, and use this solution as the sample so-
particle diameter) in series.
lution. Separately, dissolve 0.181 g of potassium sulfate in
diluted ethanol (3 in 10) to make exactly 500 mL. Pipet 5 mL
309 C.
of this solution, and add diluted ethanol (3 in 10) to make
exactly 100 mL. To 4.5 mL of this solution add 3 mL of a so-
phosphate in water to make 1000 mL. Adjust the pH of this
lution of barium chloride dihydrate (1 in 4), shake, and
solution to 3.5 with phosphoric acid.
allow to stand for 1 minute. To 2.5 mL of this solution add
Flow rate: Adjust so that the retention time of citicoline is
15 mL of the sample solution and 0.5 mL of acetic acid (31),
about 26 minutes.
and allow to stand for 5 minutes: the solution has no more
turbidity than the following control solution (not more than
150 ppm).
Control solution: Dissolve 0.181 g of potassium sulfate in
factor of the peak of citicoline are not less than 2000 and 0.9
add water to make exactly 100 mL, and proceed in the same
to 1.6, respectively.
manner as above using this solution instead of the sample so-
lution.
(3) Oxalic acid—Dissolve 0.80 g of Anhydrous Citric
Acid in 4 mL of water, add 3 mL of hydrochloric acid and
area of citicoline is not more than 1.0z.
1 g of zinc, and boil for 1 minute. After allowing to stand
Containers and storage Containers—Tight containers. for 2 minutes, take the supernatant liquid, add 0.25 mL of a
solution of phenylhydrazinium chloride (1 in 100), heat to
boil, and then cool quickly. To this solution add the equal
Anhydrous Citric Acid volume of hydrochloric acid and 0.25 mL of a solution of
potassium hexacyanoferrate (III) (1 in 20), mix, and allow to
無水クエン酸 stand for 30 minutes: the solution has no more color than
the following control solution prepared at the same time (not
more than 360 ppm expressed as oxalic anhydride).
Control solution: To 4 mL of a solution of oxalic acid di-
hydrate (1 in 10,000) add 3 mL of hydrochloric acid and 1 g
C6H8O7: 192.12
of zinc, and proceed in the same manner as the test solution.
2-Hydroxypropane-1,2,3-tricarboxylic acid (4) Heavy metals <1.07>—Proceed with 2.0 g of Anhy-
[77-92-9]
drous Citric Acid according to Method 2, and perform the
Lead Solution (not more than 10 ppm).
(5) Readily carbonizable substances—Place 1.0 g of An-
that are not harmonized are marked with symbol ( ).
hydrous Citric Acid in a Nessler tube, add 10 mL of sulfuric
acid, immediately heat in a 90 ± 19C water bath for 60
Anhydrous Citric Acid contains not less than 99.5z
minutes, and cool quickly. Compare the color of 2.0 mL
and not more than 100.5z of anhydrous citric acid
each of this solution and Matching Fluid K, using test tubes
(C6H8O7), calculated on the anhydrous basis.
12 mm in outside diameter, from a side against white back-
Description Anhydrous Citric Acid occurs as colorless ground: the solution is not more colored than the matching
crystals, white granules or crystalline powder. fluid.
It is very soluble in water, and freely soluble in ethanol
(99.5).
of Anhydrous Citric Acid, previously dried at 1059C for 2
hours, as directed in the potassium bromide disk method Assay Weigh accurately about 0.55 g of Anhydrous Citric
under Infrared Spectrophotometry <2.25>, and compare the Acid, dissolve in 50 mL of water, and titrate with 1 mol/L
spectrum with the Reference Spectrum: both spectra exhibit sodium hydroxide VS (indicator: 1 drop of phenolphthalein
similar intensities of absorption at the same wave numbers. TS).
Purity (1) Clarity and color of solution—Dissolve 2.0 g Each mL of 1 mol/L sodium hydroxide VS
of Anhydrous Citric Acid in water to make 10 mL: the solu- ＝ 64.04 mg of C6H8O7
tion is clear and colorless or has no more color than the fol- Containers and storage Containers—Tight containers.
lowing control solutions (1), (2) or (3).
CS and 6.0 mL of Iron (III) Chloride CS add diluted dilute
hydrochloric acid (1 in 10) to make 1000 mL.
JP XVII Official Monographs / Clarithromycin 717
tion of phenylhydrazinium chloride (1 in 100), heat to boil,

Citric Acid Hydrate and then cool quickly. To this solution add the equal volume
of hydrochloric acid and 0.25 mL of a solution of potassium
クエン酸水和物 hexacyanoferrate (III) (1 in 20), mix, and allow to stand for
30 minutes: the solution has no more color than the follow-
ing control solution prepared at the same time (not more
than 360 ppm expressed as oxalic anhydride).
Control solution: To 4 mL of a solution of oxalic acid di-
C6H8O7.H2O: 210.14
hydrate (1 in 10,000) add 3 mL of hydrochloric acid and 1 g
2-Hydroxypropane-1,2,3-tricarboxylic acid monohydrate
of zinc, and proceed in the same manner as the test solution.
[5949-29-1] (4) Heavy metals <1.07>—Proceed with 2.0 g of Citric
This monograph is harmonized with the European Phar- Acid Hydrate according to Method 2, and perform the test.
macopoeia and the U.S. Pharmacopeia. The parts of the text Prepare the control solution with 2.0 mL of Standard Lead
that are not harmonized are marked with symbol ( ). Solution (not more than 10 ppm).
(5) Readily carbonizable substances—Place 1.0 g of
Citric Acid Hydrate contains not less than 99.5z Citric Acid Hydrate in a Nessler tube, add 10 mL of sulfuric
and not more than 100.5z of anhydrous citric acid acid, immediately heat in a 90 ± 19C water bath for 60
(C6H8O7: 192.12), calculated on the anhydrous basis. minutes, and cool quickly. Compare the color of 2.0 mL
Description each of this solution and Matching Fluid K, using test tubes
Citric Acid Hydrate occurs as colorless crys-
12 mm in outside diameter, from a side against white back-
tals, white granules or crystalline powder.
ground: the solution is not more colored than the matching
It is very soluble in water, and freely soluble in ethanol
fluid.
(99.5).
It is efflorescent in dry air. Water <2.48> Not less than 7.5z and not more than 9.0z
of Citric Acid Hydrate, previously dried at 1059C for 2 Residue on ignition <2.44> Not more than 0.1z (1 g).
hours, as directed in the potassium bromide disk method
Assay Weigh accurately about 0.55 g of Citric Acid Hy-
drate, dissolve in 50 mL of water, and titrate <2.50> with 1
mol/L sodium hydroxide VS (indicator: 1 drop of phenol-
phthalein TS).
of Citric Acid Hydrate in water to make 10 mL: the solution
＝ 64.04 mg of C6H8O7
is clear and colorless or has no more color than the following
control solutions (1), (2) or (3). Containers and storage Containers—Tight containers.
CS and 6.0 mL of Iron (III) Chloride CS add diluted dilute
hydrochloric acid (1 in 10) to make 1000 mL. Clarithromycin
CS, 6.0 mL of Iron (III) Chloride CS and 1.0 mL of Copper クラリスロマイシン
(II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10)
to make 1000 mL.
CS, 7.2 mL of Iron (III) Chloride CS and 0.15 mL of Cop-
per (II) Sulfate CS add diluted dilute hydrochloric acid (1 in
10) to make 1000 mL.
(2) Sulfates—Dissolve 2.0 g of Citric Acid Hydrate in
water to make 30 mL, and use this solution as the sample so-
lution. Separately, dissolve 0.181 g of potassium sulfate in
diluted ethanol (3 in 10) to make exactly 500 mL. Pipet 5 mL
of this solution, and add diluted ethanol (3 in 10) to make
exactly 100 mL. To 4.5 mL of this solution add 3 mL of a so-
lution of barium chloride dihydrate (1 in 4), shake, and C38H69NO13: 747.95
allow to stand for 1 minute. To 2.5 mL of this solution add (2R,3S,4S,5R,6R,8R,10R,11R,12S,13R)-5-(3,4,6-
15 mL of the sample solution and 0.5 mL of acetic acid (31), Trideoxy-3-dimethylamino-b-D-xylo-hexopyranosyloxy)-3-
and allow to stand for 5 minutes: the solution has no more (2,6-dideoxy-3-C-methyl-3-O-methyl-a-L-ribo-
turbidity than the following control solution. (not more than hexopyranosyloxy)-11,12-dihydroxy-6-methoxy-
150 ppm). 2,4,6,8,10,12-hexamethyl-9-oxopentadecan-13-olide
Control solution: Dissolve 0.181 g of potassium sulfate in [81103-11-9]
add water to make exactly 100 mL, and proceed in the same Clarithromycin is a derivative of erythromycin.
manner as above using this solution instead of the sample so- It contains not less than 950 mg (potency) and not
lution. more than 1050 mg (potency) per mg, calculated on the
(3) Oxalic acid—Dissolve 0.80 g of Citric Acid Hydrate anhydrous basis. The potency of Clarithromycin is
in 4 mL of water, add 3 mL of hydrochloric acid and 1 g of expressed as mass (potency) of clarithromycin
zinc, and boil for 1 minute. After allowing to stand for 2 (C38H69NO13).
minutes, take the supernatant liquid, add 0.25 mL of a solu-
Description Clarithromycin occurs as a white crystalline
718 Clarithromycin / Official Monographs JP XVII
powder and has a bitter taste. AT: Peak area of each related substance obtained with the
It is soluble in acetone and in chloroform, slightly soluble sample solution
in methanol and in ethanol (95), and practically insoluble in SAT: Total area of the peaks other than clarithromycin
water. obtained with the sample solution
Identification (1) To 5 mg of Clarithromycin add 2 mL of Operating conditions—
sulfuric acid, and shake gently: a red-brown color develops. Detector, column, column temperature, mobile phase, and
(2) Dissolve 3 mg of Clarithromycin in 2 mL of acetone, flow rate: Proceed as directed in the operating conditions in
and add 2 mL of hydrochloric acid: an orange color develops the Assay.
and changes immediately to red to deep purple. Time span of measurement: About 5 times as long as the
(3) Determine the infrared absorption spectra of retention time of the main peak after 2 minutes of sample
Clarithromycin and Clarithromycin RS as directed in the injection.
potassium bromide disk method under Infrared Spectropho- System suitability—
tometry <2.25>, and compare these spectra: both spectra System performance: Proceed as directed in the system
exhibit similar intensities of absorption at the same wave suitability in the Assay.
numbers. Test for required detectability: To exactly 2 mL of the
(4) Dissolve 10 mg each of Clarithromycin and standard solution add the mobile phase to make exactly 10
Clarithromycin RS in 4 mL of chloroform, and use these mL, and use this solution as the solution for system suita-
solutions as the sample solution and standard solution. bility test. Confirm that when the procedure is run with 10
Perform the test with these solutions as directed under Thin- mL of the solution for system suitability test, the peak area
layer Chromatography <2.03>. Spot 5 mL each of the sample of clarithromycin is equivalent to 14 to 26z of that obtained
solution and standard solution on a plate of silica gel for with 10 mL of the standard solution.
thin-layer chromatography. Develop with a mixture of chlo- System repeatability: When the test is repeated 6 times
roform, methanol and ammonia water (28) (100:5:1) to a with 10 mL of the solution for system suitability test under
distance of about 15 cm, and air-dry the plate. Spray evenly the above operating conditions, the relative standard devia-
sulfuric acid on the plate, and heat at 1059C for 10 minutes: tion of the peak area of clarithromycin is not more than
the principal spot from the sample solution and the spot 3.0z.
from the standard solution show a dark purple color and
have the same R f value.
D : －87 – －979 (0.25 g calcu-
lated on the anhydrous basis, chloroform, 25 mL, 100 mm).
Assay Weigh accurately an amount of Clarithromycin and
Clarithromycin RS, equivalent to about 0.1 g (potency), and
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of dissolve in the mobile phase to make exactly 20 mL. Pipet 2
Clarithromycin according to Method 4, and perform the mL each of these solutions, add exactly 2 mL of the internal
test. Prepare the control solution with 2.0 mL of Standard standard solution, add the mobile phase to make 20 mL, and
Lead Solution (not more than 10 ppm). use these solutions as the sample solution and standard solu-
(2) Arsenic—Prepare the test solution with 1.0 g of tion. Perform the test with 10 mL each of the sample solution
Clarithromycin according to Method 3, and perform the test and standard solution as directed under Liquid Chromatog-
(not more than 2 ppm). raphy <2.01> according to the following conditions, and cal-
(3) Related substances—Weigh accurately about 0.1 g of culate the ratios, QT and QS, of the peak area of clarithromy-
Clarithromycin, dissolve in the mobile phase to make exactly cin to that of the internal standard.
Amount [ mg (potency)] of clarithromycin (C38H69NO13)
rately, weigh accurately about 10 mg of Clarithromycin RS,
＝ MS × QT/QS × 1000
dissolve in the mobile phase to make exactly 20 mL, and use
this solution as the standard solution. Perform the test with MS: Amount [mg (potency)] of Clarithromycin RS taken
cording to the following conditions, and determine the each
peak area by the automatic integration method: the amount
of each related substance calculated on the anhydrous basis
length: 210 nm).
is not more than 2.0z, and the total of them is not more
than 5.0z. Exclude any peak with an area of less than
0.05z.
Amount (z) of each related substance calculated on the Column temperature: A constant temperature of about
anhydrous basis 509C.
＝ MS/MT × AT/AS × 100 Mobile phase: A mixture of diluted 0.2 mol/L potassium
dihydrogenphosphate TS (1 in 3) and acetonitrile (13:7).
Total amount (z) of the related substances calculated on
Flow rate: Adjust so that the retention time of clarithro-
the anhydrous basis
mycin is about 8 minutes.
＝ MS/MT × SAT/AS × 100
MS: Amount (mg) of Clarithromycin RS taken System performance: When the procedure is run with 10
MT: Amount (mg) of Clarithromycin taken, calculated on mL of the standard solution under the above operating con-
the anhydrous basis ditions, clarithromycin and the internal standard are eluted
AS: Peak area of clarithromycin obtained with the stand- in this order with the resolution between these peaks being
ard solution not less than 3.
JP XVII Official Monographs / Clarithromycin Tablets 719
System repeatability: When the test is repeated 6 times and use this solution as the sample solution. Separately,
with 10 mL of the standard solution under the above operat- weigh accurately an amount of Clarithromycin RS, equiva-
ing conditions, the relative standard deviation of the ratios lent to about 28 mg (potency), and dissolve in acetonitrile for
of the peak area of clarithromycin to that of the internal liquid chromatography to make exactly 100 mL. Pipet 5 mL
standard is not more than 2.0z. of this solution, add the mobile phase to make exactly 50
ers.
the peak areas, AT and AS, of clarithromycin in each solu-
Clarithromycin Tablets tion.
クラリスロマイシン錠 Dissolution rate (z) with respect to the labeled amount
of clarithromycin (C38H69NO13)
＝ MS × AT/AS × V?/V × 1/C × 90
Clarithromycin Tablets contain not less than 93.0z
and not more than 107.0z of the labeled potency of MS: Amount [mg (potency)] of Clarithromycin RS taken
clarithromycin (C38H69NO13: 747.95). C: Labeled amount [mg (potency)] of clarithromycin
(C38H69NO13) in 1 tablet
with Clarithromycin. Operating conditions—
Identification Shake a quantity of powdered Clarithromy-
Assay.
cin Tablets, equivalent to 60 mg (potency) of Clarithromy-
cin, with 40 mL of acetone for 10 minutes, and centrifuge at
4000 rpm for 5 minutes. Evaporate 30 mL of the supernatant
liquid, and determine the infrared absorption spectrum of
the residue so obtained as directed in the potassium bromide
factor of the peak of clarithromycin are not less than 3000
disk method under Infrared Spectrophotometry <2.25>: it ex-
hibits absorption at the wave numbers of about 2980 cm－1,
2940 cm－1, 1734 cm－1, 1693 cm－1, 1459 cm－1, 1379 cm－1
and 1171 cm－1.
Uniformity of dosage units <6.02> Perform the Mass varia- area of clarithromycin is not more than 2.0z.
Assay To not less than 5 Clarithromycin Tablets add
diluted 0.2 mol/L potassium dihydrogen phosphate TS (1 in
To 1 tablet of Clarithromycin Tablets add exactly V/20
3) so that each mL contains about 8 mg (potency) of
mL of the internal standard solution (1), then add the mobile
clarithromycin (C38H69NO13), disperse to fine particles with
phase so that each mL contains about 5 mg (potency) of
the aid of ultrasonic waves, add exactly 1 mL of the internal
clarithromycin (C38H69NO13) to make V mL, and disperse to
standard solution (1) per 100 mg (potency) of clarithromy-
fine particles with the aid of ultrasonic waves for 20 minutes
cin, then add acetonitrile for liquid chromatography so that
while occasional vigorous shaking. Centrifuge this solution
each mL contains about 5 mg (potency) of clarithromycin
at 4000 rpm for 15 minutes, and filter the supernatant liquid
(C38H69NO13), and disperse to fine particles with the aid of
ultrasonic waves for 10 minutes while occasional vigorous
0.45 mm. Then, proceed as directed in the Assay.
shaking. Centrifuge of this solution at 4000 rpm for 15
Amount [mg (potency)] of clarithromycin (C38H69NO13) minutes, and filter the supernatant liquid through a mem-
＝ MS × QT/QS × V/10 brane filter with a pore size not exceeding 0.45 mm. Discard
the first 3 mL of the filtrate, to 2 mL of the subsequent fil-
MS: Amount [mg (potency)] of Clarithromycin RS taken
trate add the mobile phase to make 20 mL, and use this solu-
Internal standard solution (1)—A solution of butyl parahy- tion as the sample solution. Separately, weigh accurately an
droxybenzoate in the mobile phase (1 in 1000). amount of Clarithromycin RS, equivalent to about 50 mg
Internal standard solution (2)—To exactly 1 mL of the inter- (potency), and dissolve in the mobile phase to make exactly
nal standard solution (1) add the mobile phase to make 10 mL. Pipet 2 mL of this solution, add exactly 2 mL of the
exactly 20 mL. internal standard solution (2) and the mobile phase to make
mL of 0.05 mol/L disodium hydrogen phosphate-citric acid
buffer solution (pH 6.0) as the dissolution medium, the dis-
the ratios, QT and QS, of the peak area of clarithromycin to
solution rates in 30 minutes of a 50-mg tablet and a 200-mg
that of the internal standard.
tively. Amount [mg (potency)] of clarithromycin (C38H69NO13)
Start the test with 1 tablet of Clarithromycin Tablets, ＝ MS × QT/QS × 1/5
MS: Amount [mg (potency)] of Clarithromycin RS taken
filter with a pore size not exceeding 0.45 mm. Discard the Internal standard solution (1)—A solution of butyl parahy-
first 10 mL of the filtrate, pipet V mL of the subsequent droxybenzoate in the mobile phase (1 in 1000).
filtrate, add the mobile phase to make exactly V? mL so that Internal standard solution (2)—To exactly 1 mL of the inter-
each mL contains about 28 mg (potency) of Clarithromycin, nal standard solution (1) add the mobile phase to make
720 Clebopride Malate / Official Monographs JP XVII
exactly 20 mL. (3) Perform the test with Clebopride Malate under
Operating conditions— Flame Coloration Test <1.04> (2): a green color appears.
Purity (1) Chloride <1.03>—Dissolve 1.0 g of Clebopride
length: 210 nm).
Malate in 20 mL of acetic acid (100), add 6 mL of dilute
nitric acid and water to make 50 mL. Perform the test using
this solution as the test solution. Prepare the control solution
with 0.25 mL of 0.01 mol/L hydrochloric acid VS by adding
20 mL of acetic acid (100), 6 mL of dilute nitric acid and
509 C.
water to make 50 mL (not more than 0.009z).
Mobile phase: A mixture of diluted 0.2 mol/L potassium
(2) Heavy metals <1.07>—Proceed with 2.0 g of Cle-
dihydrogen phosphate TS (1 in 3) and acetonitrile for liquid
bopride Malate according to Method 2, and perform the
chromatography (13:7).
Flow rate: Adjust so that the retention time of clarithro-
mycin is about 8 minutes.
(3) Related substances—Dissolve 0.10 g of Clebopride
Malate in 10 mL of the mobile phase, and use this solution
as the sample solution. Pipet 0.2 mL of the sample solution,
ditions, clarithromycin and the internal standard are eluted
not less than 3.
area of both solutions by the automatic integration method:
the total area of the peaks other than clebopride obtained
the peak area of clarithromycin to that of the internal stand-
clebopride obtained from the standard solution.
Containers and storage Containers—Well-closed contain- Operating conditions—
ers. Detector: An ultraviolet absorption photometer (wave-
length: 240 nm).
Clebopride Malate ter and 25 cm in length, packed with octadecylsilanized silica
クレボプリドリンゴ酸塩 Column temperature: A constant temperature of about
259C.
Mobile phase: Dissolve 3.85 g of ammonium acetate in
water to make 500 mL, and filter through a membrane filter
with a pore size not exceeding 0.5 mm. To 400 mL of the fil-
trate add 600 mL of methanol.
Flow rate: Adjust so that the retention time of clebopride
C20H24ClN3O2.C4H6O5: 507.96
4-Amino-N-(1-benzylpiperidin-4-yl)-5-chloro-
retention time of clebopride.
2-methoxybenzamide mono-(2RS )-malate
[57645-91-7]
ard solution, and add water to make exactly 100 mL. Con-
Clebopride Malate, when dried, contains not less
firm that the peak area of clebopride obtained from 10 mL of
than 98.5z and not more than 101.0z of clebopride
this solution is equivalent to 7 to 13z of that of clebopride
malate (C20H24ClN3O2.C4H6O5).
obtained from 10 mL of the standard solution.
Description Clebopride Malate occurs as a white crystalline System performance: Dissolve 30 mg Clebopride Malate
powder. and 5 mg of propyl parahydroxybenzoate in the mobile
It is freely soluble in acetic acid (100), soluble in methanol, phase to make 100 mL. When the procedure is run with 10
sparingly soluble in water, and slightly soluble in ethanol mL of this solution under the above operating conditions,
(99.5). propyl parahydroxybenzoate and clebopride are eluted in
A solution of Clebopride Malate in methanol (1 in 25) this order with the resolution between these peaks being not
shows no optical rotation. less than 3.
solution of Clebopride Malate in methanol (1 in 80000) as di-
area of clebopride is not more than 2.5z.
both spectra exhibit similar intensities of absorption at the Loss on drying <2.41> Not more than 0.5z (1 g, 1059C,
same wavelengths. 4 hours).
(2) Determine the infrared absorption spectrum of Cle-
bopride Malate, previously dried, as directed in the potas-
sium bromide disk method under Infrared Spectrophotome- Assay Weigh accurately about 0.5 g of Clebopride Malate,
try <2.25>, and compare the spectrum with the Reference previously dried, dissolve in 30 mL of acetic acid (100), and
Spectrum: both spectra exhibit similar intensities of absorp- titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-
tion at the same wave numbers. metric titration). Perform a blank determination in the same
JP XVII Official Monographs / Clindamycin Hydrochloride 721
manner, and make any necessary correction. of Clemastine Fumarate in 10 mL of methanol by warming:
the solution is clear and colorless.
(2) Heavy metals <1.07>—Perform the test with 1.0 g of
＝ 50.80 mg of C20H24ClN3O2.C4H6O5
Clemastine Fumarate according to Method 2. Prepare the
Containers and storage Containers—Tight containers. control solution with 2.0 mL of Standard Lead Solution (not
more than 20 ppm).
(3) Arsenic <1.11>—Take 1.0 g of Clemastine Fumarate,
Clemastine Fumarate prepare the test solution according to Method 3, and per-
クレマスチンフマル酸塩 (4) Related Substances—Dissolve 0.10 g of Clemastine
Fumarate in 5 mL of methanol, and use this solution as the
the standard solution (1). Pipet 5 mL of the standard solu-
tion (1), add methanol to make exactly 10 mL, and use this
solution as the standard solution (2). Perform the test with
C21H26ClNO.C4H4O4: 459.96 phy <2.03>. Spot 5 mL each of the sample solution and stand-
(2R)-2-{2-[(1R)-1-(4-Chlorophenyl)-1- ard solutions (1) and (2) on a plate of silica gel for thin-layer
phenylethoxy]ethyl}-1-methylpyrrolidine monofumarate chromatography. Develop the plate with a mixture of chlo-
[14976-57-9] roform, methanol and ammonia solution (28) (90:10:1) to a
distance of about 15 cm, and air-dry the plate. After
Clemastine Fumarate, when dried, contains not less spraying evenly Dragendorff 's TS on the plate, immediately
than 98.5z of clemastine fumarate (C21H26ClNO. spray evenly hydrogen peroxide TS: the spots other than the
C4H4O4). principal spot from the sample solution are not more intense
than the spot from the standard solution (1), and not more
Description Clemastine Fumarate occurs as a white, crys-
than 2 spots from the sample solution are more intense than
talline powder. It is odorless.
the spot from the standard solution (2).
It is sparingly soluble in methanol and in acetic acid (100),
slightly soluble in ethanol (95), very slightly soluble in Loss on drying <2.41> Not more than 0.5z (1 g, 1059C,
diethyl ether, and practically insoluble in water. 4 hours).
Identification (1) To 5 mg of Clemastine Fumarate add 5 Residue on ignition <2.44> Not more than 0.2z (1 g).
mL of sulfuric acid, and shake to dissolve: a yellow color de-
Assay Weigh accurately about 0.4 g of Clemastine
velops. Slowly drop this solution into 10 mL of water: the
Fumarate, previously dried, dissolved in 50 mL of acetic acid
yellow color immediately disappears.
(100), and titrate <2.50> with 0.1 mol/L perchloric acid VS
(2) To 0.01 g of Clemastine Fumarate add 1 mL of fum-
(potentiometric titration). Perform a blank determination,
ing nitric acid, and evaporate on a water bath to dryness.
Then add 2 mL of diluted hydrochloric acid (1 in 2) and
0.2 g of zinc powder, heat for 10 minutes on a water bath, Each mL of 0.1 mol/L perchloric acid VS
cool, and filter. Add 20 mL of water to the filtrate. The solu- ＝ 46.00 mg of C21H26ClNO.C4H4O4
tion responds to the Qualitative Tests <1.09> for primary aro-
matic amines.
(3) To 5 mL of a solution of Clemastine Fumarate (1 in
50,000), add 5 mL of 4-dimethylaminobenzaldehyde TS, and
warm for 10 minutes: a red-purple color develops. Clindamycin Hydrochloride
(4) Perform the test with Clemastine Fumarate as di-
クリンダマイシン塩酸塩
appears.
(5) Dissolve 0.04 g of Clemastine Fumarate and 0.01 g of
fumaric acid for thin-layer chromatography in 2 mL each of
a mixture of ethanol (95) and water (4:1) by gentle warming,
and use these solutions as the sample solution and the stand-
ard solution, respectively. Perform the test with these solu-
on a plate of silica gel with fluorescent indicator for thin- C18H33ClN2O5S.HCl: 461.44
layer chromatography. Develop the plate with a mixture of Methyl 7-chloro-6,7,8-trideoxy-6-[(2S,4R)-1-methyl-4-
isopropyl ether, formic acid and water (90:7:3) to a distance propylpyrrolidine-2-carboxamido]-1-thio-L-threo-a-D-
of about 10 cm, and air-dry the plate. Examine the plate galacto-octopyranoside monohydrochloride
under ultraviolet light (main wavelength: 254 nm): the spot [21462-39-5]
with larger R f value from the sample solution has the same
R f value as the spot from the standard solution. Clindamycin Hydrochloride is the hydrochloride of
a derivative of lincomycin.
＋16 – ＋189 (after drying,
D:
Melting point <2.60> 176 – 1809C (with decomposition). anhydrous basis. The potency of Clindamycin Hydro-
chloride is expressed as mass (potency) of clindamycin
722 Clindamycin Hydrochloride Capsules / Official Monographs JP XVII
(C18H33ClN2O5S: 424.98). Assay Weigh accurately an amount of Clindamycin Hydro-

chloride and Clindamycin Hydrochloride RS, equivalent to
Description Clindamycin Hydrochloride occurs as white to
about 20 mg (potency), dissolve each in the mobile phase to
grayish white, crystals or crystalline powder.
make exactly 20 mL, and use these solutions as the sample
It is freely soluble in water and in methanol, and slightly
solution and the standard solution, respectively. Perform the
soluble in ethanol (99.5).
Identification (1) Determine the infrared absorption spec- standard solution as directed under Liquid Chromatography
trum of Clindamycin Hydrochloride as directed in the potas- <2.01> according to the following conditions, and determine
sium chloride disk method under Infrared Spectrophotome- the peak areas, AT and AS, of clindamycin in each solution.
Amount [mg (potency)] of clindamycin (C18H33ClN2O5S)
Spectrum or the spectrum of Clindamycin Hydrochloride
＝ MS × AT/AS × 1000
the same wave numbers. MS: Amount [mg (potency)] of Clindamycin Hydrochlo-
(2) A solution of Clindamycin Hydrochloride (1 in 100) ride RS taken
responds to the Qualitative Tests <1.09> (2) for chloride.
D : ＋135 – ＋1509(0.5 g calcu- Detector: An ultraviolet absorption photometer (wave-
lated on the anhydrous basis, water, 25 mL, 100 mm). length: 210 nm).
Clindamycin Hydrochloride according to Method 4, and
259C.
Mobile phase: To 0.05 mol/L potassium dihydrogen phos-
phate TS add 8 mol/L potassium hydroxide TS to adjust the
sample solution, add the mobile phase to make exactly 100
pH to 7.5. To 550 mL of this solution add 450 mL of aceto-
nitrile for liquid chromatography.
Flow rate: Adjust so that the retention time of clindamy-
peak area of clindamycin B, having the relative retention
time of about 0.7 to clindamycin, and that of 7-epiclindamy-
cin, having the relative retention time of about 0.8 to clin-
factor of the peak of clindamycin are not less than 6000 and
damycin, obtained from the sample solution are not larger
than 2 times the peak area of clindamycin obtained from the
standard solution, the area of the peak other than clindamy-
cin and the peaks mentioned above from the sample solution
is not larger than the peak area of clindamycin from the
area of clindamycin is not more than 1.0z.
clindamycin from the sample solution is not larger than 4 Containers and storage Containers—Tight containers.
times the peak area of clindamycin from the standard solu-
tion.
Operating conditions— Clindamycin Hydrochloride
flow rate: Proceed as directed in the operating conditions in Capsules
the Assay.
クリンダマイシン塩酸塩カプセル
retention time of clindamycin, beginning after the solvent
peak. Clindamycin Hydrochloride Capsules contain not
System suitability— less than 93.0z and not more than 107.0z of the
Test for required detectability: Pipet 1 mL of the standard labeled potency of clindamycin (C18H33ClN2O5S:
solution, and add the mobile phase to make exactly 10 mL. 424.98).
Confirm that the peak area of clindamycin obtained from 20
mL of this solution is equivalent to 7 to 13z of that of clin-
sules, with Clindamycin Hydrochloride.
damycin obtained from 20 mL of the standard solution.
System performance: When the procedure is run with 20 Identification To an amount of the contents of Clindamy-
mL of the standard solution under the above operating con- cin Hydrochloride Capsules, equivalent to 10 mg (potency)
ditions, the number of theoretical plates and the symmetry of Clindamycin Hydrochloride, add 2 mL of methanol,
factor of the peak of clindamycin are not less than 6000 and shake well, centrifuge, and use the supernatant liquid as the
not more than 1.5, respectively. sample solution. Separately, dissolve 10 mg of Clindamycin
System repeatability: When the test is repeated 6 times Hydrochloride RS in 2 mL of methanol, and use this solu-
with 20 mL of the standard solution under the above operat- tion as the standard solution. Perform the test with these so-
ing conditions, the relative standard deviation of the peak lutions as directed under Thin-layer Chromatography <2.03>.
area of clindamycin is not more than 2.0z. Spot 10 mL each of the sample solution and standard solu-
tion on a plate of silica gel for thin-layer chromatography.
Develop the plate with a mixture of methanol, toluene and
ammonia solution (28) (140:60:3) to a distance of about 12
JP XVII Official Monographs / Clindamycin Hydrochloride Capsules 723
cm, and air-dry the plate. Spray evenly a mixture of 500 mL mL of the standard solution under the above operating con-
of a solution of L-tartaric acid (1 in 5) and 50 mL of bismuth ditions, the number of theoretical plates and the symmetry
subnitrate TS on the plate: the R f values of the principal factor of the peak of clindamycin are not less than 3000 and
spot with the sample solution and the spot with the standard not more than 2.0, respectively.
solution are not different each other. System repeatability: When the test is repeated 6 times
To 1 capsule of Clindamycin Hydrochloride Capsules add Assay Take out the contents of not less than 20 Clindamy-
a suitable amount of the mobile phase, shake for 30 minutes, cin Hydrochloride Capsules, weigh accurately the mass of
and add the mobile phase to make exactly V mL so that each the contents, and powder. Weigh accurately a portion of the
mL contains 0.75 mg (potency) of Clindamycin Hydrochlo- powder, equivalent to about 75 mg (potency) of Clindamycin
ride. Centrifuge this solution, and use the supernatant liquid Hydrochloride, add the mobile phase, shake for 30 minutes,
as the sample solution. Then, proceed as directed in the and add the mobile phase to make exactly 100 mL. Centri-
Assay. fuge this solution, and use the supernatant liquid as the
Amount [mg (potency)] of clindamycin (C18H33ClN2O5S)
Clindamycin Hydrochloride RS, equivalent to about 75 mg
＝ MS × AT/AS × V/100
(potency), dissolve in the mobile phase to make exactly 100
MS: Amount [mg (potency)] of Clindamycin Hydrochlo- mL, and use this solution as the standard solution. Perform
ride RS taken the test with exactly 20 mL each of the sample solution and
the peak areas, AT and AS, of clindamycin in each solution.
dissolution rate of a 75-mg capsule in 15 minutes and that of Amount [mg (potency)] of clindamycin (C18H33ClN2O5S)
a 150-mg capsule in 30 minutes are not less than 80z, ＝ M S × AT / AS
respectively.
MS: Amount [mg (potency)] of Clindamycin Hydrochlo-
Start the test with 1 capsule of Clindamycin Hydrochlo-
ride RS taken
ride Capsules, withdraw not less than 20 mL of the medium
at the specified minute after starting the test, and filter Operating conditions—
through a membrane filter with a pore size not exceeding Detector: An ultraviolet absorption photometer (wave-
0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL length: 210 nm).
of the subsequent filtrate, add water to make exactly V? so Column: A stainless steel column 4.6 mm in inside diame-
that each mL contains about 83 mg (potency) of Clindamycin ter and 15 cm in length, packed with octadecylsilanized silica
Hydrochloride, and use this solution as the sample solution. gel for liquid chromatography (5 mm in particle diameter).
Separately, weigh accurately an amount of Clindamycin Hy- Column temperature: A constant temperature of about
drochloride RS, equivalent to about 17 mg (potency), dis- 409C.
solve in water to make exactly 200 mL, and use this solution Mobile phase: To 0.05 mol/L of potassium dihydrogen
as the standard solution. Perform the test with exactly 20 mL phosphate TS add 8 mol/L potassium hydroxide TS to
each of the sample solution and standard solution as directed adjust the pH to 7.5. To 550 mL of this solution add 450 mL
under Liquid Chromatography <2.01>, and determine the of acetonitrile for liquid chromatography.
peak areas, AT and AS, of clindamycin in each solution. Flow rate: Adjust so that the retention time of clindamy-
of clindamycin (C18H33ClN2O5S)
＝ MS × AT/AS × V?/V × 1/C × 450
MS: Amount [mg (potency)] of Clindamycin Hydrochlo- ditions, the number of theoretical plates and the symmetry
ride RS taken factor of the peak of clindamycin are not less than 3000 and
C: Labeled amount [mg (potency)] of clindamycin not more than 2.0, respectively.
(C18H33ClN2O5S) in 1 tablet System repeatability: When the test is repeated 6 times
length: 210 nm).
Column: A stainless steel column 4.6 mm in inside diame- Containers and storage Containers—Tight containers.
409 C.
Mobile phase: Adjust the pH of 0.05 mol/L potassium di-
hydrogen phosphate TS to 7.5 with 8 mol/L potassium hy-
droxide TS. To 550 mL of this solution add 450 mL of aceto-
nitrile.
724 Clindamycin Phosphate / Official Monographs JP XVII
area of the peaks other than clindamycin phosphate from the
Clindamycin Phosphate sample solution is not larger than 4 times the peak area of
clindamycin phosphate from the standard solution.
クリンダマイシンリン酸エステル Operating conditions—
the Assay.
retention time of clindamycin phosphate, beginning after the
solvent peak.
C18H34ClN2O8PS: 504.96
Methyl 7-chloro-6,7,8-trideoxy-6-[(2S,4R)-1-methyl-4-
propylpyrrolidine-2-carboxamido]-1-thio-L-threo-a-D-
actly 10 mL. Confirm that the peak area of clindamycin
galacto-octopyranoside 2-dihydrogen phosphate
phosphate obtained from 20 mL of this solution is equivalent
[24729-96-2]
to 7 to 13z of that obtained from 20 mL of the standard so-
lution.
Clindamycin Phosphate is a derivative of clindamy-
cin. Water <2.48> Not more than 6.0z (0.5 g, volumetric titra-
It contains not less than 800 mg (potency) and not tion, direct titration).
Assay Weigh accurately an amount of Clindamycin Phos-
anhydrous basis. The potency of Clindamycin Phos-
phate and Clindamycin Phosphate RS, equivalent to about
phate is expressed as mass (potency) of clindamycin
20 mg (potency), add exactly 25 mL of the internal standard
(C18H33ClN2O5S: 424.98).
solution and the mobile phase to make 100 mL, and use
Description Clindamycin Phosphate occurs as a white to these solutions as the sample solution and standard solution.
pale yellowish white crystalline powder. Perform the test with 20 mL each of the sample solution and
It is freely soluble in water, sparingly soluble in methanol, standard solution as directed under Liquid Chromatography
and practically insoluble in ethanol (95). <2.01> according to the following conditions, and calculate
the ratios, QT and QS, of the peak area of clindamycin phos-
phate to that of the internal standard.
of Clindamycin Phosphate, previously dried at 1009C for 2
hours, as directed in the paste method under Infrared Spec- Amount [ mg (potency)] of clindamycin (C18H33ClN2O5S)
trophotometry <2.25>, and compare the spectrum with the ＝ MS × QT/QS × 1000
Reference Spectrum or the spectrum of Clindamycin Phos-
MS: Amount [mg (potency)] of Clindamycin Phosphate
phate RS previously dried at 1009C for 2 hours: both spectra
RS taken
numbers. Internal standard solution—A solution of methyl parahy-
＋115 – ＋1309(0.25 g calcu-
D:
pH <2.54> Dissolve 0.10 g of Clindamycin Phosphate in 10 length: 210 nm).
mL of water. The pH of the solution is between 3.5 and 4.5. Column: A stainless steel column 4 mm in inside diameter
and 25 cm in length, packed with octylsilanized silica gel for
of Clindamycin Phosphate in 10 mL of freshly boiled and
cooled water: the solution is clear and colorless.
259C.
(2) Heavy metals <1.07>—Proceed with 2.0 g of Clin-
damycin Phosphate according to Method 4, and perform the
phosphate in 775 mL of water, adjust the pH to 2.5 with
phosphoric acid, and add 225 mL of acetonitrile.
cin phosphate is about 8 minutes.
of Clindamycin Phosphate according to Method 4, and per-
(4) Related substances—Dissolve 0.1 g of Clindamycin
Phosphate in 100 mL of the mobile phase, and use this solu-
ditions, clindamycin phosphate and the internal standard are
of the peak area of clindamycin phosphate to that of the in-
peak area by the automatic integration method: the peak
ternal standard is not more than 2.5z.
area of clindamycin, having the relative retention time of
about 1.8 to clindamycin phosphate, obtained from the sam- Containers and storage Containers—Tight containers.
ple solution is not larger than 1/2 times the peak area of clin-
damycin phosphate from the standard solution, and the total
JP XVII Official Monographs / Clinofibrate 725
Clindamycin Phosphate Injection Clinofibrate

クリンダマイシンリン酸エステル注射液クリノフィブラート
Clindamycin Phosphate Injection is an aqueous in-

jection.
110.0z of the labeled potency of clindamycin phos-
phate (C18H34ClN2O8PS: 504.96).
C28H36O6: 468.58
tions, with Clindamycin Phosphate.
2,2?-(4,4?-Cyclohexylidenediphenoxy)-2,2?-
Description Clindamycin Phosphate Injection is a clear, dimethyldibutanoic acid
colorless or light yellow liquid. [30299-08-2]
Identification To a volume of Clindamycin Phosphate
Clinofibrate, when dried, contains not less than
Injection, equivalent to 0.15 g (potency) of Clindamycin
98.5z of clinofibrate (C28H36O6).
Phosphate, add 4 mL of water, 2 mL of 8 mol/L sodium hy-
droxide TS and 0.1 mL of sodium pentacyanonitrosylferrate Description Clinofibrate occurs as a white to yellowish
(III) TS, mix, heat in a water bath for 10 minutes, and add 2 white powder. It is odorless and has no taste.
mL of hydrochloric acid: a blue-green color develops. It is freely soluble in methanol, in ethanol (99.5), in ace-
tone and in diethyl ether, and practically insoluble in water.
A solution of Clinofibrate in methanol (1 in 20) shows no
optical rotation.
pH <2.54> 6.0 – 7.0 Melting point: about 1469C (with decomposition).
Bacterial endotoxins <4.01> Less than 0.1 EU/mg (po- Identification (1) Determine the absorption spectrum of a
tency). solution of Clinofibrate in ethanol (99.5) (1 in 50,000) as
Foreign insoluble matter <6.06> Perform the test according both spectra exhibit similar intensities of absorption at the
to Method 1: it meets the requirement. same wavelengths.
Clinofibrate, previously dried, as directed in the potassium
ment.
Sterility <4.06> Perform the test according to the Mem- <2.25>, and compare the spectrum with the Reference Spec-
brane filtration method: it meets the requirement. trum: both spectra exhibit similar intensities of absorption at
Assay Measure exactly a volume of Clindamycin Phos-
phate Injection, equivalent to about 0.3 g (potency) of Clin- Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
damycin Phosphate, and add the mobile phase to make ex- Clinofibrate according to Method 2, and perform the test.
actly 100 mL. Pipet 7 mL of this solution, add exactly 25 mL Prepare the control solution with 2.0 mL of Standard Lead
of the internal standard solution and the mobile phase to Solution (not more than 20 ppm).
make 100 mL, and use this solution as the sample solution. (2) Arsenic <1.11>—Prepare the test solution with 1.0 g
Separately, weigh accurately an amount of Clindamycin of Clinofibrate according to Method 3, and perform the test
Phosphate RS, equivalent to about 20 mg (potency), dissolve (not more than 2 ppm).
in exactly 25 mL of the internal standard solution, add the (3) Related substances—Dissolve 0.10 g of Clinofibrate
mobile phase to make 100 mL, and use this solution as the in 10 mL of acetone, and use this solution as the sample so-
standard solution. Then, proceed as directed in the Assay lution. Pipet 1 mL of the sample solution, and add acetone
under Clindamycin Phosphate. to make exactly 50 mL. Pipet 5 mL of this solution, add ace-
tone to make exactly 20 mL, and use this solution as the
Amount [mg (potency)] of clindamycin phosphate
(C18H34ClN2O8PS)
＝ MS × QT/QS × 100/7
MS: Amount [mg (potency)] of Clindamycin Phosphate plate of silica gel with fluorescent indicator for thin-layer
RS taken chromatography. Develop the plate with a mixture of chlo-
roform, cyclohexane and acetic acid (100) (12:5:3) to a dis-
ultraviolet light (main wavelength: 254 nm): the spots other
Containers and storage Containers—Hermetic containers. than the principal spot from the sample solution are not
um, 609C, 3 hours).
Isomer ratio To 50 mg of Clinofibrate add 0.4 mL of
726 Clobetasol Propionate / Official Monographs JP XVII
thionyl chloride, stopper tightly, heat on a water bath of It gradually turns yellow by light.
609 C for 5 minutes with occasional shaking, and evaporate Melting point: about 1969 C (with decomposition).
the excess thionyl chloride at a temperature not exceeding
Identification Determine the infrared absorption spectra of
609 C under reduced pressure. Dissolve the residue in 2 mL
Clobetasol Propionate as directed in the paste method under
of toluene previously dried with synthetic zeolite for drying,
add 2 mL of a solution of D-(＋)-a-methylbenzylamine in
trum with the Reference Spectrum or the spectrum of
toluene previously dried with synthetic zeolite for drying
Clobetasol Propionate RS: both spectra exhibit similar in-
(3 in 100), mix gently, allow to stand for 10 minutes, and
tensities of absorbance at the same wave numbers.
evaporate the toluene at a temperature not exceeding 609C
under reduced pressure. Dissolve the residue in 5 mL of chlo- Optical rotation <2.49> [a]20
roform, and use this solution as the sample solution. Per- 0.1 g, methanol, 10 mL, 100 mm).
form the test with 5 mL of the sample solution as directed
Clobetasol Propionate according to Method 2, and perform
lowing conditions. Determine each peak area, Aa, Ab and Ac,
of three peaks appear in order near the retention time of 40
minutes: a value, Ab/(Aa ＋ Ab ＋ Ac) × 100, is between 40
(2) Related substances—Dissolve 10 mg of Clobetasol
and 70.
Propionate in 100 mL of the mobile phase, and use this solu-
length: 254 nm).
Column: A stainless steel column about 4 mm in inside di-
ameter and about 30 cm in length, packed with silica gel for
peak area of these solutions by the automatic integration
209 C.
method: the area of the peak other than clobetasol
Mobile phase: A mixture of hexane and 2-propanol
propionate obtained from the sample solution is not larger
(500:3).
than 2/5 times the peak area of clobetasol propionate ob-
Flow rate: Adjust so that the retention time of the peak
tained from the standard solution. Furthermore, the total
appearing first is about 35 minutes.
area of the peaks other than clobetasol propionate from the
Selection of column: Proceed with 5 mL of the sample so-
sample solution is not larger than the peak area of clobetasol
lution under the above operating conditions. Use a column
propionate from the standard solution.
giving a complete separation of the three peaks.
Assay Weigh accurately about 0.45 g of Clinofibrate, pre- Detector, column, column temperature, mobile phase, and
viously dried, dissolve in 40 mL of ethanol (99.5), add 30 mL flow rate: Proceed as directed in the operating conditions in
of water, and titrate <2.50> with 0.1 mol/L sodium hydrox- the Assay.
ide VS (indicator: 3 drops of phenolphthalein TS). Perform Time span of measurement: About 2.5 times as long as the
a blank determination, and make any necessary correction. retention time of clobetasol propionate, beginning after the
solvent peak.
＝ 23.43 mg of C28H36O6
Containers and storage Containers—Tight containers. solution, and add the mobile phase to make exactly 50 mL.
Confirm that the peak area of clobetasol propionate ob-
tained from 10 mL of this solution is equivalent to 2.8 to
Clobetasol Propionate 5.2z of that obtained from 10 mL of the standard solution.
System performance: Dissolve 20 mg of Clobetasol
クロベタゾールプロピオン酸エステル Propionate in 20 mL of methanol. To 5 mL of this solution
add 10 mL of a solution of beclometasone dipropionate in
methanol (1 in 1000), and then add the mobile phase to make
50 mL. When the procedure is run with 10 mL of this solu-
tion under the above conditions, clobetasol propionate and
beclometasone dipropionate are eluted in this order with the
with 10 mL of the standard solution under the above condi-
C25H32ClFO5: 466.97 tions, the relative standard deviation of the peak area of
21-Chloro-9-fluoro-11b,17-dihydroxy- clobetasol propionate is not more than 2.0z.
16b-methylpregna-1,4-diene-3,20-dione 17-propanoate
[25122-46-7]
3 hours).
Clobetasol Propionate, when dried, contains not Residue on ignition <2.44> Not more than 0.1z (1 g, plati-
less than 97.0z and not more than 102.0z of num crucible).
clobetasol propionate (C25H32ClFO5).
Assay Weigh accurately about 10 mg each of Clobetasol
Description Clobetasol Propionate occurs as a white to Propionate and Clobetasol Propionate RS, both previously
pale yellowish white crystalline powder. dried, dissolve each in the mobile phase, add exactly 100 mL
It is soluble in methanol and in ethanol (99.5), and practi- of the internal standard solution, add the mobile phase to
cally insoluble in water. make 250 mL, and use these solutions as the sample solution
JP XVII Official Monographs / Clocapramine Hydrochloride Hydrate 727
and standard solution. Perform the test with 10 mL each of chloride (C28H37ClN4O.2HCl: 553.99).
Description Clocapramine Hydrochloride Hydrate occurs
as white, crystals or crystalline powder. It is odorless, and
has a bitter taste.
area of clobetasol propionate to that of the internal stand-
ard.
water and in methanol, slightly soluble in ethanol (95), in
Amount (mg) of clobetasol propionate (C25H32ClFO5) chloroform and in isopropylamine, and practically insoluble
＝ M S × QT / QS in acetic anhydride and in diethyl ether.
MS: Amount (mg) of Clobetasol Propionate RS taken
Melting point: about 2609C (with decomposition, after
Internal standard solution—A solution of beclometasone drying).
dipropionate in the mobile phase (1 in 5000).
Identification (1) To 5 mL of a solution of Clocapramine
Hydrochloride Hydrate (1 in 2500) add 1 mL of nitric acid: a
blue color develops at first, and rapidly changes to deep
length: 240 nm).
blue, and then changes to green to yellow-green.
Clocapramine Hydrochloride Hydrate in methanol (1 in
tometry <2.24>,, and compare the spectrum with the Refer-
259 C.
phate dihydrate in 900 mL of water, adjust the pH to 2.5
with phosphoric acid, and then add water to make 1000 mL.
Clocapramine Hydrochloride Hydrate as directed in the po-
To 425 mL of this solution add 475 mL of acetonitrile and
tassium bromide disk method under Infrared Spectropho-
100 mL of methanol.
Flow rate: Adjust so that the retention time of clobetasol
ence Spectrum: both spectra exhibit similar intensities of
propionate is about 10 minutes.
(4) Dissolve 0.1 g of Clocapramine Hydrochloride Hy-
drate in 10 mL of water by warming, and after cooling, add
mL of the standard solution under the above conditions,
2 mL of ammonia TS, and filter. Acidify the filtrate with
clobetasol propionate and the internal standard are eluted in
dilute nitric acid: the solution responds to the Qualitative
Tests <1.09> (2) for chloride.
less than 8.
System repeatability: When the test is repeated 6 times Purity (1) Sulfate <1.14>—Dissolve 0.5 g of Clocapra-
with 10 mL of the standard solution under the above condi- mine Hydrochloride Hydrate in 40 mL of water by warming,
tions, the relative standard deviation of the ratio of the peak after cooling, and add 1 mL of dilute hydrochloric acid and
area of clobetasol propionate to that of the internal standard water to make 50 mL. Perform the test using this solution as
is not more than 1.0z. the test solution. Prepare the control solution with 0.50 mL
of 0.005 mol/L sulfuric acid VS (not more than 0.048z).
Clocapramine Hydrochloride Hydrate according to Method
2, and perform the test. Prepare the control solution with 2.0
Clocapramine Hydrochloride (3) Related substances—Conduct this procedure without
Hydrate exposure to light, using light-resistant vessels. Dissolve 0.10
g of Clocapramine Hydrochloride Hydrate in 10 mL of a
クロカプラミン塩酸塩水和物 mixture of chloroform and isopropylamine (99:1), and use
ple solution, add a mixture of chloroform and isopropyla-
mine (99:1) to make exactly 100 mL, and use this solution as
chromatography. Develop the plate with a mixture of diethyl
ether, ethyl acetate, methanol and ammonia solution (28)
C28H37ClN4O.2HCl.H2O: 572.01 nm): the spots other than the principal spot from the sample
1?-[3-(3-Chloro-10,11-dihydro-5H-dibenz[b, f ]azepin-5- solution are not more intense than the spot from the stand-
yl)propyl]-1,4?-bipiperidine-4?-carboxamide ard solution.
dihydrochloride monohydrate
Loss on drying <2.41> 2.0 – 3.5z (0.5 g, in vacuum at a
[60789-62-0]
pressure not exceeding 0.67 kPa, phosphorus (V) oxide,
1059C, 4 hours).
Clocapramine Hydrochloride Hydrate, when dried,
contains not less than 98.0z of clocapramine hydro- Residue on ignition <2.44> Not more than 0.1z (1 g).
728 Clofedanol Hydrochloride / Official Monographs JP XVII
Assay Weigh accurately about 0.5 g of Clocapramine Hy- lowing conditions. Determine each peak area of both solu-
drochloride Hydrate, previously dried, dissolve in 70 mL of tions by the automatic integration method: the total area of
a mixture of acetic anhydride and acetic acid (100) (6:1), and the peaks other than clofedanol from the sample solution is
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- not larger than the peak area of clofedanol from the stand-
metric titration). Perform a blank determination, and make ard solution.
any necessary correction. Operating conditions—
length: 220 nm).
＝ 27.70 mg of C28H37ClN4O.2HCl
Containers and storage Containers—Tight containers. diameter and about 15 cm in length, packed with octadecyl-
Storage—Light-resistant. silanized silica gel for liquid chromatography (5 mm in parti-
cle diameter).
Clofedanol Hydrochloride 409C.
Mobile phase: Dissolve 1.34 g of potassium methanesul-
クロフェダノール塩酸塩 fonate in diluted phosphoric acid (1 in 1000) to make 1000
mL, and to 650 mL of this solution add 350 mL of metha-
nol.
Flow rate: Adjust so that the retention time of clofedanol
is about 9 minutes.
Selection of column: Dissolve 0.01 g each of Clofedanol
Hydrochloride and ethyl parahydroxybenzoate in methanol
to make 100 mL. Proceed with 3 mL of this solution under
the above operating conditions, and calculate the resolution.
C17H20ClNO.HCl: 326.26
Use a column giving elution of clofedanol and ethyl parahy-
(1RS )-1-(2-Chlorophenyl)-3-dimethylamino-1-
droxybenzoate in this order with the resolution of these
phenylpropan-1-ol monohydrochloride
[511-13-7]
that the peak height of clofedanol obtained from 3 mL of the
Clofedanol Hydrochloride, when dried, contains
standard solution composes between 20z and 50z of the
not less than 98.5z of clofedanol hydrochloride
full scale.
(C17H20ClNO.HCl).
Time span of measurement: About three times as long as
Description Clofedanol Hydrochloride occurs as white, the retention time of clofedanol, beginning after the solvent
crystals or crystalline powder. peak.
It is freely soluble in methanol, in ethanol (95) and in
acetic acid (100), sparingly soluble in water, and practically
um, silica gel, 809C, 3 hours).
insoluble in diethyl ether.
A solution of Clofedanol Hydrochloride in methanol Residue on ignition <2.44> Not more than 0.1z (1 g).
(1 in 20) does not show optical rotation.
Assay Weigh accurately about 0.5 g of Clofedanol Hydro-
Melting point: about 1909 C (after drying, with decompo-
sition).
(100), add 35 mL of acetic anhydride, and titrate <2.50> with
Identification (1) Determine the absorption spectrum of a 0.1 mol/L perchloric acid VS (potentiometric titration). Per-
solution of Clofedanol Hydrochloride in 0.01 mol/L hydro- form a blank determination, and make any necessary correc-
chloric acid TS (1 in 2500) as directed under Ultraviolet- tion.
＝ 32.63 mg of C17H20ClNO.HCl
(2) Determine the infrared absorption spectrum of Containers and storage Containers—Tight containers.
Clofedanol Hydrochloride, previously dried, as directed in
photometry <2.25>, and compare the spectrum with the Ref- Clofibrate
absorption at the same wave numbers. クロフィブラート
(3) A solution of Clofedanol Hydrochloride (1 in 100)
Clofedanol Hydrochloride according to Method 2, and per-
Standard Lead Solution (not more than 10 ppm). C12H15ClO3: 242.70
(2) Related substances—Dissolve 0.05 g of Clofedanol Ethyl 2-(4-chlorophenoxy)-2-methylpropanoate
Hydrochloride in 25 mL of methanol, and use this solution [637-07-0]
add methanol to make exactly 100 mL, and use this solution Clofibrate contains not less than 98.0z of
as the standard solution. Perform the test with exactly 3 mL clofibrate (C12H15ClO3), calculated on the unhydrous
each of the sample solution and standard solution as directed basis.
JP XVII Official Monographs / Clofibrate Capsules 729
Description Clofibrate occurs as a colorless or light yellow, standard: QT is not greater than QS.
clear, oily liquid. It has a characteristic odor and taste, Internal standard solution—A solution of 4-ethoxyphenol in
which is bitter at first, and subsequently sweet. the mobile phase (1 in 30,000).
It is miscible with methanol, with ethanol (95), with Operating conditions—
ethanol (99.5), with diethyl ether and with hexane, and prac- Detector: An ultraviolet absorption photometer (wave-
tically insoluble in water. length: 275 nm).
It is gradually decomposed by light. Column: A stainless steel column about 4 mm in inside di-
ameter and about 30 cm in length, packed with cyanopropyl-
silanized silica gel for liquid chromatography (5 to 10 mm in
solution of Clofibrate in ethanol (99.5) (1 in 10,000) as
particle diameter).
and compare the spectrum with the Reference Spectrum 1 or
259C.
the spectrum of a solution of Clofibrate RS prepared in the
Mobile phase: A mixture of hexane, 2-propanol and acetic
acid (100) (1970:30:1).
Flow rate: Adjust so that the retention time of clofibrate is
Separately, determine the absorption spectrum of a solution
about 2 minutes.
of Clofibrate in ethanol (99.5) (1 in 100,000) as directed
Selection of column: Dissolve 10.0 g of Clofibrate, 6 mg
of 4-chlorophenol and 6 mg of 4-ethoxyphenol in 1000 mL
compare the spectrum with the Reference Spectrum 2 or the
of hexane. Proceed with 20 mL of this solution under the
spectrum of a solution of Clofibrate RS prepared in the same
above operating conditions, and calculate the resolution.
Use a column giving elution of clofibrate, 4-chlorophenol
and 4-ethoxyphenol in this order, with the resolution be-
tween the peaks of clofibrate and 4-chlorophenol is not less
Clofibrate as directed in the liquid film method under In-
than 5, and with the resolution between the peaks of 4-chlo-
frared Spectrophotometry <2.25>, and compare the spectrum
rophenol and 4-ethoxyphenol is not less than 2.0.
with the Reference Spectrum or the spectrum of Clofibrate
RS: both spectra exhibit similar intensities of absorption at Water <2.48> Not more than 0.2z (5 g, volumetric titra-
the same wave numbers. tion, direct titration).
Refractive index <2.45> n 20
D : 1.500 – 1.505 Residue on ignition <2.44> Not more than 0.1z (1 g).
20: 1.137 – 1.144 Assay Weigh accurately about 0.5 g of Clofibrate, add
exactly 50 mL of 0.1 mol/L potassium hydroxide-ethanol
Purity (1) Acidity—Dissolve 2.0 g of Clofibrate in 100
VS, and heat in a water bath under a reflux condenser with a
mL of neutralized ethanol, and add 1 drop of phenolphthal-
carbon dioxide absorbing tube (soda-lime) for 2 hours with
ein TS and 0.20 mL of 0.1 mol/L sodium hydroxide VS: the
frequent shaking. Cool, and titrate <2.50> immediately the
solution is red in color.
excess potassium hydroxide with 0.1 mol/L hydrochloric
acid VS (indicator: 3 drops of phenolphthalein TS). Perform
Clofibrate according to Method 2, and perform the test. Pre-
a blank determination.
lution (not more than 10 ppm). Each mL of 0.1 mol/L potassium hydroxide-ethanol VS
(3) Arsenic <1.11>—To 5.0 g of Clofibrate add 20 mL of ＝ 24.27 mg of C12H15ClO3
nitric acid and 5 mL of sulfuric acid, and heat until white
fumes are evolved. After cooling, if necessary, add further 5
mL of nitric acid, heat until white fumes are evolved, and
repeat this procedure until the solution is colorless to light
yellow. After cooling, add 15 mL of saturated ammonium
oxalate solution, and heat again until white fumes are Clofibrate Capsules
evolved. Cool, add water to make 25 mL, use 5 mL of this
クロフィブラートカプセル
solution as the test solution, and perform the test.
Color standard: Prepare a solution according to the above
procedure without using Clofibrate as the blank. Transfer 5 Clofibrate Capsules contain not less than 93.0z and
mL of the solution to a generator bottle, add 2.0 mL of not more than 107.0z of the labeled amount of
Standard Arsenic Solution, and then proceed as directed in clofibrate (C12H15ClO3: 242.70).
the test solution (not more than 20 ppm).
(4) p-Chlorophenol—To 1.0 g of Clofibrate add exactly
sules, with Clofibrate.
1 mL of the internal standard solution, then add the mobile
phase to make 5 mL, and use this solution as the sample Identification Cut and open Clofibrate Capsules, and use
solution. Separately, dissolve 10 mg of 4-chlorophenol in a the contents as the sample. Determine the absorption
mixture of hexane and 2-propanol (9:1) to make exactly 100 spectrum of a solution of the sample in ethanol (99.5) (1 in
mL. Pipet 10 mL of this solution, and add a mixture of 10,000) as directed under Ultraviolet-visible Spectropho-
hexane and 2-propanol (9:1) to make exactly 50 mL. Pipet 6 tometry <2.24>: it exhibits a maximum between 278 nm and
mL of this solution, add exactly 4 mL of the internal stand- 282 nm, and it exhibits a maximum between 224 nm and 228
ard solution, then add the mobile phase to make 20 mL, and nm after diluting this solution 10 times with ethanol (99.5).
Purity p-Chlorophenol—Cut and open not less than 20
Clofibrate Capsules, and proceed with 1.0 g of the well-
mixed contents as directed in the Purity (4) under Clofibrate.
Internal standard solution—A solution of 4-ethoxyphenol in
QS, of the peak area of 4-chlorophenol to that of the internal
730 Clomifene Citrate / Official Monographs JP XVII
the mobile phase (1 in 30,000). than 98.0z of clomifene citrate (C26H28ClNO.
C6H8O7).
Assay Weigh accurately not less than 20 Clofibrate Cap-
sules, cut and open the capsules, rinse the inside of the cap- Description Clomifene Citrate occurs as a white to pale yel-
sules with a small amount of diethyl ether after taking out lowish white powder. It is odorless.
the contents, evaporate the diethyl ether by allowing the cap- It is freely soluble in methanol and in acetic acid (100),
sules to stand at room temperature, and weigh the capsules sparingly soluble in ethanol (95), and practically insoluble in
accurately. Weigh accurately an amount of the contents, diethyl ether.
equivalent to about 0.1 g of clofibrate (C12H15ClO3), dissolve It gradually changes in color by light.
in acetonitrile to make exactly 100 mL. Pipet 5 mL of this Melting point: about 1159 C
solution, add exactly 5 mL of the internal standard solution,
Identification (1) To 2 mL of a solution of Clomifene
Citrate in methanol (1 in 200) add 2 mL of Reinecke salt TS:
weigh accurately about 0.1 g of Clofibrate RS, proceed in
a light red precipitate is produced.
the same manner as directed for the sample solution, and use
the solution so obtained as the standard solution. Perform
Clomifene Citrate in 0.1 mol/L hydrochloric acid TS (1 in
ence Spectrum or the spectrum of a solution of Clomifene
ratios, QT and QS, of the peak area of clofibrate to that of
Citrate RS prepared in the same manner as the sample solu-
Amount (mg) of clofibrate (C12H15ClO3) the same wavelengths.
＝ M S × Q T / QS (3) A solution of Clomifene Citrate in methanol (1 in
200) responds to the Qualitative Tests <1.09> (1) and (2) for
MS: Amount (mg) of Clofibrate RS taken
citrate salt.
Internal standard solution—A solution of ibuprofen in the
Purity (1) Clarity and color of solution—A solution of
1.0 g of Clomifene Citrate in 30 mL of methanol is clear and
colorless.
(2) Heavy metals <1.07>—Proceed with 2.0 g of Clomi-
length: 275 nm).
fene Citrate according to Method 2, and perform the test.
silanized silica gel for liquid chromatography (5 to 10 mm in
particle diameter). Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-
Column temperature: A constant temperature of about um, phosphorus (V) oxide, 3 hours).
259 C.
Mobile phase: A mixture of acetonitrile and diluted phos-
phoric acid (1 in 1000) (3:2). Isomer ratio To 10 mg of Clomifene Citrate add 10 mL of
Flow rate: Adjust so that the retention time of clofibrate is water and 1 mL of sodium hydroxide TS, and shake to uni-
about 10 minutes. formly disperse. Add 10 mL of ethyl acetate, shake vigor-
Selection of column: Dissolve 0.05 g of clofibrate and ously for 5 minutes, allow to stand for 5 minutes, and use
0.3 g of ibuprofen in 50 mL of acetonitrile. Proceed with 10 the upper layer as the sample solution. Perform the test with
mL of this solution under the above operating conditions, 1 mL of the sample solution as directed under Gas Chroma-
and calculate the resolution. Use a column giving elution of tography <2.02> according to the following conditions. De-
ibuprofen and clofibrate in this order with the resolution be- termine the areas of two adjacent peaks, Aa and Ab, having
tween these peaks being not less than 6. the retention time of about 8 minutes, where Aa is the peak
area of shorter retention time and Ab is the peak area of lon-
ger retention time: Ab/(Aa ＋ Ab) is between 0.3 and 0.5.
ers.
and 15 m in length, coated the inside surface with a layer
Clomifene Citrate about 0.1 mm thick of dimethylpolysiloxane for gas chroma-
tography.
クロミフェンクエン酸塩
2309C.
about 2709C.
3009C.
Flow rate: Adjust so that the retention time of the first
C26H28ClNO.C6H8O7: 598.08
peak of clomifene citrate is about 7.5 minutes.
2-[4-(2-Chloro-1,2-diphenylvinyl)phenoxy]-N, N-
Split ratio: 1:50.
diethylethylamine monocitrate
[50-41-9]
of the sample solution under the above operating conditions,
Clomifene Citrate, when dried, contains not less
the resolution between the two adjacent peaks having the
JP XVII Official Monographs / Clomipramine Hydrochloride 731
retention time of about 8 minutes is not less than 5. minute after starting the test, and filter through a membrane
System repeatability: When the test is repeated 6 times filter with a pore size not exceeding 0.45 mm. Discard the
with 1 mL of the sample solution under the above operating first 10 mL of the filtrate, pipet V mL of the subsequent fil-
conditions, the relative standard deviation of the result of trate, add the dissolution medium to make exactly V? mL so
Ab/(Aa ＋ Ab) is not more than 1.0z. that each mL contains about 28 mg of clomifene citrate
(C26H28ClNO.C6H8O7), and use this solution as the sample
Assay Weigh accurately about 1 g of Clomifene Citrate,
solution. Separately, weigh accurately about 28 mg of
previously dried, dissolve in 50 mL of acetic acid (100), and
Clomifene Citrate RS, previously dried in vacuum using
titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2
phosphorus (V) oxide as a desiccant for 3 hours, and dissolve
drops of crystal violet TS). Perform a blank determination,
in methanol to make exactly 50 mL. Pipet 5 mL of this solu-
tion, add the dissolution medium to make exactly 100 mL,
Each mL of 0.1 mol/L perchloric acid VS and use this solution as the standard solution. Determine the
＝ 59.81 mg of C26H28ClNO.C6H8O7 absorbances, AT and AS, at 291 nm of the sample solution
and standard solution as directed under Ultraviolet-visible
Spectrophotometry <2.24>, using the dissolution medium as
the blank.
Clomifene Citrate Tablets of clomifene citrate (C26H28ClNO.C6H8O7)
＝ MS × AT/AS × V?/V × 1/C × 90
クロミフェンクエン酸塩錠
MS: Amount (mg) of Clomifene Citrate RS taken
C: Labeled amount (mg) of clomifene citrate
Clomifene Citrate Tablets contain not less than (C26H28ClNO.C6H8O7) in 1 tablet
Assay Weigh accurately, and powder not less than 20
amount of the clomifene citrate (C26H28ClNO.C6H8O7:
Clomifene Citrate Tablets. Weigh accurately a portion of
598.08).
the powder, equivalent to about 50 mg of clomifene citrate
Method of preparation Prepare as directed under Tablets, (C26H28ClNO.C6H8O7), add 50 mL of methanol, shake for
with Clomifene Citrate. 10 minutes, and add methanol to make exactly 100 mL. Cen-
trifuge a portion of this solution, pipet 4 mL of the superna-
Identification Weigh a portion of powdered Clomifene
tant liquid, add methanol to make exactly 100 mL, and use
Citrate Tablets, equivalent to 50 mg of Clomifene Citrate,
shake vigorously with 50 mL of methanol for 10 minutes,
rately about 50 mg of Clomifene Citrate RS, previously
centrifuge, and use the supernatant liquid as the sample solu-
dried in a desiccator (in vacuum, phosphorus (V) oxide) for
tion. Separately, dissolve 10 mg of Clomifene Citrate RS in
3 hours, and dissolve in methanol to make exactly 100 mL.
10 mL of methanol, and use this solution as the standard
Pipet 4 mL of this solution, and dilute with methanol to
solution. Determine the absorbances, AT and AS, of the sam-
of the sample solution and standard solution on a plate of
ple solution and the standard solution, respectively, at 295
nm as directed under Ultraviolet-visible Spectrophotometry
raphy. Develop the plate with a mixture of 2-propanol, tolu-
<2.24>.
ene and diethylamine (10:10:1) to a distance of about 10 cm,
and air-dry the plate. Examine under ultraviolet light (main Amount (mg) of clomifene citrate (C26H28ClNO.C6H8O7)
wavelength: 254 nm): the spots from the sample solution and ＝ MS × AT/AS
standard solution show the same R f value.
To 1 tablet of Clomifene Citrate Tablets add 10 mL of
water, and shake until the tablets are disintegrated. To this Clomipramine Hydrochloride
solution add 50 mL of methanol, shake for 10 minutes, and
クロミプラミン塩酸塩
add methanol to make exactly 100 mL. Centrifuge this solu-
tion, pipet 4 mL of the supernatant liquid, add methanol to
make exactly V mL so that each mL contains about 20 mg of
clomifene citrate (C26H28ClNO.C6H8O7), and use this solu-
tion as the sample solution. Proceed as directed in the Assay.
Amount (mg) of clomifene citrate (C26H28ClNO.C6H8O7)
＝ MS × AT/AS × V/100
C19H23ClN2.HCl: 351.31
Dissolution <6.10> When the test is performed at 50 revolu- 3-(3-Chloro-10,11-dihydro-5H-dibenz[b, f ]azepin-
tions per minute according to the Paddle method, using 900 5-yl)-N, N-dimethylpropylamine monohydrochloride
mL of 1st fluid for dissolution test as the dissolution me- [17321-77-6]
dium, the dissolution rate in 30 minutes of Clomifene Citrate
Tablets is not less than 80z. Clomipramine Hydrochloride, when dried, contains
Start the test with 1 tablet of Clomifene Citrate Tablets, not less than 98.5z of clomipramine hydrochloride
withdraw not less than 20 mL of the medium at the specified (C19H23ClN2.HCl).
732 Clonazepam / Official Monographs JP XVII
Description Clomipramine Hydrochloride occurs as a Loss on drying <2.41> Not more than 0.5z (1 g, 1059C,
white to pale yellow, crystalline powder. It is odorless. 3 hours).
water, in methanol and in chloroform, soluble in ethanol
(95), sparingly soluble in acetic anhydride, slightly soluble in Assay Weigh accurately about 0.5 g of Clomipramine Hy-
acetone, and practically insoluble in ethyl acetate and in drochloride, previously dried, dissolve in 50 mL of a mixture
diethyl ether. of acetic anhydride and acetic acid (100) (7:3), and titrate
Identification (1) Dissolve 3 mg of Clomipramine Hydro-
chloride in 1 mL of nitric acid: a deep blue color develops.
Clomipramine Hydrochloride in 0.1 mol/L hydrochloric Each mL of 0.1 mol/L perchloric acid VS
acid TS (3 in 100,000) as directed under Ultraviolet-visible ＝ 35.13 mg of C19H23ClN2.HCl
ers.
(3) Take 1 g of Clomipramine Hydrochloride in a sepa-
rator, dissolve in 10 mL of water, add 5 mL of sodium hy-
droxide TS, and extract with two 30-mL portions of diethyl
ether [the water layer is used for Identification (4)]. Combine Clonazepam
the diethyl ether extracts, add 20 mL of water, and shake.
クロナゼパム
Take diethyl ether layer, dry with a small portion of anhy-
drous sodium sulfate, and filter. Evaporate the combined ex-
tracts by warming on a water bath, and proceed the test with
the residue as directed under Flame Coloration Test <1.04>
(2): a green color appears.
(4) The solution neutralized by adding dilute nitric acid
to the water layer obtained in (3) responds to the Qualitative
Tests <1.09> for chloride.
C15H10ClN3O3: 315.71
pH <2.54> Dissolve 1.0 g of Clomipramine Hydrochloride
5-(2-Chlorophenyl)-7-nitro-1,3-dihydro-2H-1,4-
in 10 mL of water: the pH of this solution is between 3.5 and
benzodiazepin-2-one
5.0.
[1622-61-3]
Clonazepam, when dried, contains not less than
99.0z of clonazepam (C15H10ClN3O3).
of Clomipramine Hydrochloride in 10 mL of water: the solu-
tion is clear and colorless to pale yellow. Description Clonazepam occurs as white to light yellow,
(2) Heavy metals <1.07>—Proceed with 2.0 g of crystals or crystalline powder.
Clomipramine Hydrochloride according to Method 2, and It is sparingly soluble in acetic anhydride and in acetone,
perform the test. Prepare the control solution with 2.0 mL of slightly soluble in methanol and in ethanol (95), very slightly
Standard Lead Solution (not more than 10 ppm). soluble in diethyl ether, and practically insoluble in water.
(3) Arsenic <1.11>—Prepare the test solution with 1.0 g It is gradually colored by light.
of Clomipramine Hydrochloride according to Method 3, and Melting point: about 2409 C (with decomposition).
(4) Related substances—Dissolve 0.20 g of Clomipra-
solution of Clonazepam in methanol (1 in 100,000) as di-
mine Hydrochloride in 10 mL of methanol, and use this so-
lution as the sample solution. Separately, weigh 20 mg of
Imipramine Hydrochloride, dissolve in methanol to make
same wavelengths.
tion (1). Then pipet 1 mL of the sample solution, and add
methanol to make exactly 50 mL. Pipet 5 mL of the solu-
Clonazepam, previously dried, as directed in the potassium
tion, add methanol to make exactly 50 mL, and use this solu-
tion as the standard solution (2). Perform the test with these
solutions (1) and (2) on a plate of silica gel for thin-layer
(3) Perform the test with Clonazepam as directed under
chromatography. Develop the plate with a mixture of ethyl
acetate, acetone and ammonia solution (28) (15:5:1) to a dis-
tance of about 10 cm, and air-dry the plate. Spray evenly po- Purity (1) Chloride <1.03>—To 1.0 g of Clonazepam add
tassium dichromate-sulfuric acid TS on the plate: the spot 50 mL of water, allow to stand for 1 hour with occasional
from the sample solution, corresponding to that from the shaking, and filter. Discard the first 20 mL portion of the fil-
standard solution (1), is not more intense than the spot from trate, take the subsequent 20 mL portion of the filtrate, and
the standard solution (1). Each of the spots other than the add 6 mL of dilute nitric acid and water to make 50 mL. Use
principal spot and the spot mentioned above from the sam- this solution as the test solution, and perform the test. Pre-
ple solution is not more intense than the spot from the stand- pare the control solution as follows: to 0.25 mL of 0.01
ard solution (2). mol/L hydrochloric acid VS add 6 mL of dilute nitric acid
JP XVII Official Monographs / Clonazepam Tablets 733
(2) Heavy metals <1.07>—Proceed with 1.0 g of tion, add a mixture of methanol and water (7:3) to make ex-
Clonazepam according to Method 4, and perform the test. actly 100 mL, and use this solution as the standard solution.
Prepare the control solution with 2.0 mL of Standard Lead Perform the test with exactly 15 mL each of the sample solu-
Solution (not more than 20 ppm). tion and standard solution as directed under Liquid Chroma-
(3) Related substances—Dissolve 0.25 g of Clonazepam tography <2.01> according to the following conditions, and
in 10 mL of acetone, and use this solution as the sample so- determine the peak areas, AT and AS, of clonazepam in each
lution. Pipet 1 mL of the sample solution, add acetone to solution.
make exactly 100 mL, then pipet 1 mL of this solution, add
Amount (mg) of clonazepam (C15H10ClN3O3)
acetone to make exactly 10 mL, and use this solution as the
＝ MS × AT/AS × 3/25
directed under Thin-layer Chromatography <2.03>. Spot 10 MS: Amount (mg) of clonazepam for assay taken
length: 310 nm).
nitromethane and acetone (10:1) to a distance of about 12
cm, and air-dry the plate. Examine under ultraviolet light
(main wavelength: 254 nm): the spots other than the princi-
pal spot from the sample solution are not more intense than
the spot from the standard solution.
259C.
C, Mobile phase: A mixture of water, acetonitrile and metha-
4 hours). nol (4:3:3).
Flow rate: Adjust so that the retention time of clonazepam
is about 5 minutes.
Assay Weigh accurately about 0.5 g of Clonazepam, previ- System suitability—
ously dried, dissolve in 70 mL of acetic anhydride, and System performance: When the procedure is run with 15
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- mL of the standard solution under the above operating con-
metric titration). Perform a blank determination, and make ditions, the number of theoretical plates and the symmetry
any necessary correction. factor of the peak of clonazepam are not less than 3000 and
＝ 31.57 mg of C15H10ClN3O3
Containers and storage Containers—Well-closed containing conditions, the relative standard deviation of the peak
ers. area of clonazepam is not more than 1.0z.
Clonazepam Fine Granules

クロナゼパム細粒 Clonazepam Tablets
クロナゼパム錠
Clonazepam Fine Granules contain not less than
Clonazepam Tablets contain not less than 95.0z
amount of clonazepam (C15H10ClN3O3: 315.71).
Method of preparation Prepare as directed under Gran- clonazepam (C15H10ClN3O3: 315.71).
ules, with Clonazepam.
Identification Powder Clonazepam Fine Granules. To a with Clonazepam.
portion of the powder, equivalent to 1 mg of Clonazepam,
Identification Powder Clonazepam Tablets. To a portion
add an appropriate volume of methanol and shake for 10
of the powder, equivalent to 1 mg of Clonazepam, add an
minutes, add methanol to make 100 mL, and filter. Deter-
appropriate volume of methanol and shake for 10 minutes,
mine the absorption spectrum of the filtrate as directed
then add methanol to make 100 mL, and filter. Determine
under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-
the absorption spectrum of the filtrate as directed under
its a maximum between 307 nm and 311 nm.
Dissolution Being specified separately when the drug is maximum between 307 nm and 311 nm.
Assay Powder Clonazepam Fine Granules. Weigh accu- ing to the following method: it meets the requirement of the
rately a portion of the powder, equivalent to about 2.4 mg of Content uniformity test.
clonazepam (C15H10ClN3O3), add exactly 30 mL of a mixture To 1 tablet of Clonazepam Tablets, add V/10 mL of
of methanol and water (7:3), and shake for 15 minutes. Cen- methanol, shake for 15 minutes, add 2-propanol to make
trifuge this solution, pipet 5 mL of the supernatant liquid, exactly V mL so that each mL contains about 10 mg of
add a mixture of methanol and water (7:3) to make exactly clonazepam (C15H10ClN3O3). Filter this solution through a
20 mL, and use this solution as the sample solution. Sepa- membrane filter with a pore size not exceeding 0.45 mm. Dis-
rately, weigh accurately about 20 mg of clonazepam for card the first 10 mL of the filtrate, and use the subsequent
assay, previously dried at 1059 C for 4 hours, dissolve in filtrate as the sample solution. Separately, weigh accurately
methanol to make exactly 50 mL. Pipet 5 mL of this solu- about 20 mg of clonazepam for assay, previously dried at
734 Clonidine Hydrochloride / Official Monographs JP XVII
1059C for 4 hours, dissolve in methanol to make exactly 200 solution. Perform the test with exactly 10 mL each of the
mL. Pipet 10 mL of this solution, add 2-propanol to make sample solution and standard solution as directed under
exactly 100 mL, and use this solution as the standard solu- Liquid Chromatography <2.01> according to the following
tion. Determine the absorbances, AT and AS, at 312 nm of conditions, and determine the peak areas, AT and AS, of
the sample solution and standard solution as directed under clonazepam in each solution.
Ultraviolet-visible Spectrophotometry <2.24>, using a mix-
ture of 2-propanol and methanol (9:1) as the control.
＝ MS × AT/AS × 1/10
MS: Amount (mg) of clonazepam for assay taken
＝ MS × AT/AS × V/2000
Dissolution <6.10> When the test is performed at 50 revolu- length: 310 nm).
tions per minute according to the Paddle method, using 900 Column: A stainless steel column 4.6 mm in inside diame-
mL of water as the dissolution medium, the dissolution rate ter and 15 cm in length, packed with octadecylsilanized silica
in 30 minutes of 0.5-mg tablet and 1-mg tablet is not less gel for liquid chromatography (5 mm in particle diameter).
than 80z, and that of 2-mg tablet is not less than 75z. Column temperature: A constant temperature of about
Start the test with 1 tablet of Clonazepam Tablets, with- 259C.
draw not less than 20 mL of the medium at the specified Mobile phase: A mixture of water, acetonitrile and metha-
minute after starting the test, and filter through a membrane nol (4:3:3).
filter with a pore size not exceeding 0.45 mm. Discard the Flow rate: Adjust so that the retention time of clonazepam
first 10 mL of the filtrate, pipet V mL of the subsequent fil- is about 5 minutes.
trate, add water to make exactly V? mL so that each mL con- System suitability—
tains about 0.56 mg of clonazepam (C15H10ClN3O3), and use System performance: When the procedure is run with 10
this solution as the sample solution. Separately, weigh accu- mL of the standard solution under the above operating con-
rately about 22 mg of clonazepam for assay, previously dried ditions, the number of theoretical plates and the symmetry
at 1059 C for 4 hours, and dissolve in methanol to make ex- factor of the peak of clonazepam are not less than 3000 and
actly 100 mL. Pipet 5 mL of this solution, and add methanol not more than 1.5, respectively.
to make exactly 50 mL. Pipet 5 mL of this solution and add System repeatability: When the test is repeated 6 times
water to make exactly 200 mL, and use this solution as the with 10 mL of the standard solution under the above operat-
standard solution. Perform the test with exactly 100 mL each ing conditions, the relative standard deviation of the peak
of the sample solution and standard solution as directed area of clonazepam is not more than 1.0z.
of clonazepam in each solution.
of clonazepam (C15H10ClN3O3) Clonidine Hydrochloride
＝ MS × AT/AS × V?/V × 1/C × 9/4
クロニジン塩酸塩
C: Labeled amount (mg) of clonazepam (C15H10ClN3O3)
in 1 tablet
Assay.
C9H9Cl2N3.HCl: 266.55
2-(2,6-Dichlorophenylimino)imidazolidine
monohydrochloride
[4205-91-8]
factor of the peak of clonazepam are not less than 2000 and
Clonidine Hydrochloride, when dried, contains
not less than 99.0z of clonidine hydrochloride
(C9H9Cl2N3.HCl).
ing conditions, the relative standard deviation of the peak Description Clonidine Hydrochloride occurs as white, crys-
area of clonazepam is not more than 2.0z. tals or crystalline powder.
It is freely soluble in methanol, soluble in water and in
ethanol (95), slightly soluble in acetic acid (100), and practi-
Clonazepam Tablets, and powder. Weigh accurately a por-
cally insoluble in acetic anhydride and in diethyl ether.
tion of the powder, equivalent to about 2.5 mg of clonazep-
am (C15H10ClN3O3), add exactly 50 mL of a mixture of meth- Identification (1) To 5 mL of a solution of Clonidine Hy-
anol and water (7:3), and shake for 15 minutes. Centrifuge drochloride (1 in 1000) add 6 drops of Dragendorff's TS: an
this solution, and use the supernatant liquid as the sample orange precipitate is formed.
solution. Separately, weigh accurately about 25 mg of (2) Determine the absorption spectrum of a solution of
clonazepam for assay, previously dried at 1059C for 4 hours, Clonidine Hydrochloride in 0.01 mol/L hydrochloric acid
dissolve in methanol to make exactly 25 mL. Pipet 5 mL of TS (3 in 10,000) as directed under Ultraviolet-visible Spectro-
this solution, add a mixture of methanol and water (7:3) to photometry <2.24>, and compare the spectrum with the Ref-
make exactly 100 mL, and use this solution as the standard erence Spectrum: both spectra exhibit similar intensities of
JP XVII Official Monographs / Cloperastine Hydrochloride 735

(3) Determine the infrared absorption spectrum of Cloperastine Hydrochloride
Clonidine Hydrochloride, previously dried, as directed in the
potassium chloride disk method under Infrared Spectropho- クロペラスチン塩酸塩
(4) A solution of Clonidine Hydrochloride (1 in 50) re-
pH <2.54> Dissolve 1.0 g of Clonidine Hydrochloride in 20
C20H24ClNO.HCl: 366.32
of Clonidine Hydrochloride in 20 mL of water: the solution
1-{2-[( RS )-(4-Chlorophenyl)(phenyl)methoxy]ethyl}piperidine
monohydrochloride
(2) Heavy metals <1.07>—Proceed with 2.0 g of Cloni-
[14984-68-0]
Cloperastine Hydrochloride, when dried, contains
not less than 98.5z of cloperastine hydrochloride
(C20H24ClNO.HCl).
of Clonidine Hydrochloride according to Method 3, and per-
form the test (not more than 4 ppm). Description Cloperastine Hydrochloride occurs as white,
(4) Related substances—Dissolve 0.20 g of Clonidine crystals or crystalline powder.
Hydrochloride in 2 mL of methanol, and use this solution as It is very soluble in water, in methanol, in ethanol (95) and
the sample solution. Pipet 1 mL of the sample solution, and in acetic acid (100), and soluble in acetic anhydride.
add methanol to make exactly 100 mL. Pipet 1 mL and 2 mL A solution of Cloperastine Hydrochloride (1 in 10) shows
of this solution, to each add methanol to make exactly 20 no optical rotation.
mL, and use these solutions as the standard solution (1) and
the standard solution (2), respectively. Perform the test with
solution of Cloperastine Hydrochloride in 0.1 mol/L hydro-
chloric acid TS (1 in 2500) as directed under Ultraviolet-
phy <2.03>. Spot 2 mL each of the sample solution and stand-
ard solutions (1) and (2) on a plate of silica gel for thin-layer
with the Reference Spectrum 1: both spectra exhibit similar
chromatography. Develop the plate with a mixture of tolu-
intensities of absorption at the same wavelengths. Sepa-
ene, 1,4-dioxane, ethanol (99.5) and ammonia solution (28)
rately, determine the absorption spectrum of a solution of
(10:8:2:1) to a distance of about 12 cm, air-dry the plate,
Cloperastine Hydrochloride in 0.1 mol/L hydrochloric acid
and then dry at 1009C for 1 hour. Spray evenly sodium
TS (1 in 62,500) as directed under Ultraviolet-visible Spectro-
hypochlorite TS on the plate, air-dry the plate for 15
minutes, and then spray evenly potassium iodide starch TS
erence Spectrum 2: both spectra exhibit similar intensities of
on the plate: the spots other than the principal spot and the
spot of the starting point from the sample solution are not
more intense than the spot from the standard solution (2),
Cloperastine Hydrochloride, previously dried, as directed in
and the numbers of spots other than the principal spot and
the spot of the starting point, which are more intense than
the spot from the standard solution (1), are not more than 3.
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, absorption at the same wave numbers.
4 hours). (3) Shake 10 mL of a solution of Cloperastine Hydro-
chloride (1 in 100) with 2 mL of ammonia TS and 20 mL of
diethyl ether, separate the water layer, wash the water layer
Assay Weigh accurately about 0.4 g of Clonidine Hydro- with 20 mL of diethyl ether, and filter. Acidify the filtrate
chloride, previously dried, and dissolve in 30 mL of acetic with dilute nitric acid: the solution responds to the Qualita-
acid (100) by warming. After cooling, add 70 mL of acetic tive Tests <1.09> for chloride.
anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid
VS (potentiometric titration). Perform a blank determina-
tion, and make any necessary correction. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
Cloperastine Hydrochloride according to Method 2, and per-
＝ 26.66 mg of C9H9Cl2N3.HCl
Containers and storage Containers—Tight containers. (2) Related substances—Dissolve 40 mg of Cloperastine
736 Clopidogrel Sulfate / Official Monographs JP XVII
method: The areas of two peaks corresponding to the rela-
tive retention times about 0.8 and about 3.0 to cloperastine Clopidogrel Sulfate
obtained from the sample solution are not larger than the
peak area of cloperastine obtained from the standard solu- クロピドグレル硫酸塩
tion, respectively, and the area of the peak corresponding to
the relative retention time about 2.0 is not larger than 5/3
times the peak area of cloperastine from the standard solu-
tion, and the areas of the peaks other than cloperastine and
the peaks mentioned above from the sample solution are not
larger than 3/5 times the peak area of cloperastine from the
standard solution. The total area of these peaks is not larger
than 2 times the peak area of cloperastine from the standard C16H16ClNO2S.H2SO4: 419.90
solution. Methyl (2S)-2-(2-chlorophenyl)-2-[6,7-dihydrothieno[3,2-c]pyridin-
Operating conditions— 5(4H)-yl]acetate monosulfate
Detector: An ultraviolet absorption photometer (wave- [120202-66-6]
length: 222 nm).
Column: A stainless steel column about 5 mm in inside Clopidogrel Sulfate contains not less than 97.0z
diameter and about 15 cm in length, packed with octadecyl- and not more than 101.5z of clopidogrel sulfate
silanized silica gel for liquid chromatography (5 mm in parti- (C16H16ClNO2S.H2SO4), calculated on the anhydrous
cle diameter). basis.
Description Clopidogrel Sulfate occurs as a white to pale
259 C.
yellowish white, crystalline powder or powder.
Mobile phase: A mixture of methanol, 0.1 mol/L mono-
It is freely soluble in water and in methanol, and soluble in
basic potassium phosphate TS and perchloric acid
ethanol (99.5).
(500:250:1).
It gradually develops a brown color on exposure to light.
Flow rate: Adjust so that the retention time of cloper-
astine is about 7 minutes.
Selection of column: Dissolve 0.03 g of Cloperastine Hy-
drochloride and 0.04 g of benzophenone in 100 mL of the Identification (1) Determine the absorption spectrum of a
mobile phase. To 2.0 mL of this solution add the mobile solution of Clopidogrel Sulfate in methanol (3 in 10,000) as
phase to make 50 mL. Perform the test with 20 mL of this so- directed under Ultraviolet-visible Spectrophotometry <2.24>,
lution under the above operating conditions, and calculate and compare the spectrum with the Reference Spectrum or
the resolution. Use a column giving elution of cloperastine the spectrum of a solution of Clopidogrel Sulfate RS pre-
and benzophenone in this order with the resolution between pared in the same manner as the sample solution: both spec-
these peaks being not less than 6. tra exhibit similar intensities of absorption at the same wave-
Detection sensitivity: Adjust the detection sensitivity so lengths.
that the peak height of cloperastine obtained from 20 mL of (2) Determine the infrared absorption spectrum of
the standard solution is about 30z of the full scale. Clopidogrel Sulfate as directed in the potassium bromide
Time span of measurement: About 4 times as long as the disk method under Infrared Spectrophotometry <2.25>, and
retention time of cloperastine, beginning after the solvent compare the spectrum with the Reference Spectrum or the
peak. spectrum of Clopidogrel Sulfate RS: both spectra exhibit
If any difference appears between the spectra, dissolve
3 hours).
Clopidogrel Sulfate, or each of Clopidogrel Sulfate and
Residue on ignition <2.44> Not more than 0.1z (1 g). Clopidogrel Sulfate RS in ethanol (99.5), respectively. Then
evaporate the ethanol to dryness, and repeat the test on the
Assay Weigh accurately about 0.5 g of Cloperastine Hy-
residues dried in vacuum.
(3) Perform the test with Clopidogrel Sulfate as directed
under Flame Coloration Test <1.04> (2): a green color ap-
pears.
(4) A solution of Clopidogrel Sulfate in a mixture of
water and methanol (1:1) (1 in 100) responds to the Qualita-
Each mL of 0.1 mol/L perchloric acid VS tive Tests <1.09> (1) for sulfate.
＝ 36.63 mg of C20H24ClNO.HCl
Containers and storage Containers—Tight containers. Clopidogrel Sulfate according to Method 4, and perform the
Storage—Light-resistant. test. Prepare the control solution with 2.0 mL of Standard
(2) Related substances—Dissolve 65 mg of Clopidogrel
Sulfate in 10 mL of a mixture of acetonitrile for liquid chro-
matography and mobile phase A (3:2), and use this solution
and add a mixture of acetonitrile for liquid chromatography
and the mobile phase A (3:2) to make exactly 100 mL. Pipet
2.5 mL of this solution, add a mixture of acetonitrile for
liquid chromatography and the mobile phase A (3:2) to
JP XVII Official Monographs / Clopidogrel Sulfate 737
solution. Perform the test with exactly 10 mL each of the termine each peak area by the automatic integration method:
sample solution and standard solution as directed under the peak area of the optical isomer, having the relative reten-
Liquid Chromatography <2.01> according to the following tion time of about 0.6 to clopidogrel, obtained from the
conditions, and determine each peak area by the automatic sample solution is not larger than the peak area of
integration method: the area of the peak, having the relative clopidogrel obtained from the standard solution.
retention time of about 0.5 and about 1.1 to clopidogrel, Operating conditions—
obtained from the sample solution is not larger than 2 times Detector: An ultraviolet absorption photometer (wave-
the peak area of clopidogrel obtained from the standard length: 220 nm).
solution, the area of the peak other than clopidogrel and the Column: A stainless steel column 4.6 mm in inside diame-
peaks mentioned above from the sample solution is not ter and 25 cm in length, packed with cellulose derivative-
larger than the peak area of clopidogrel from the standard bonded silica gel for liquid chromatography (10 mm in parti-
solution, and the total area of the peaks other than cle diameter).
clopidogrel from the sample solution is not larger than 5 Column temperature: A constant temperature of about
times the peak area of clopidogrel from the standard 259C.
solution. Mobile phase: A mixture of heptane for liquid chromatog-
Operating conditions— raphy and ethanol (99.5) for liquid chromatography (17:3).
Detector, column and column temperature: Proceed as di- Flow rate: Adjust so that the retention time of clopidogrel
rected in the operating conditions in the Assay. is about 18 minutes.
Mobile phase A: Dissolve 0.87 g of sodium 1-pen- System suitability—
tanesufonate in 1000 mL of water, and adjust to pH 2.5 with System performance: When the procedure is run with 10
phosphoric acid. To 950 mL of this solution add 50 mL of mL of the standard solution under the above operating con-
methanol. ditions, the number of theoretical plates and the symmetry
Mobile phase B: A mixture of acetonitrile for liquid chro- factor of the peak of clopidogrel are not less than 3500 and
matography and methanol (19:1). not more than 2.0, respectively.
Flowing of mobile phase: Control the gradient by mixing System repeatability: When the test is repeated 6 times
the mobile phases A and B as directed in the following table. with 10 mL of the standard solution under the above operat-
Time after injection Mobile phase A Mobile phase B area of clopidogrel are not more than 2.0z.
of sample (min) (volz) (volz) Water <2.48> Not more than 0.5z (1 g, coulometric titra-
tion).
0– 3 89.5 10.5
3 – 48 89.5 → 31.5 10.5 → 68.5 Residue on ignition <2.44> Not more than 0.1z (1 g).
48 – 68 31.5 68.5
Assay Weigh accurately about 45 mg each of Clopidogrel
Sulfate and Clopidogrel Sulfate RS (separately, determine
Flow rate: 1.0 mL per minute. the water <2.48> in the same manner as Clopidogrel Sulfate),
Time span of measurement: For 68 minutes after injec- and dissolve them separately in the mobile phase to make ex-
tion, beginning after the solvent peak. actly 50 mL. Take exactly 7 mL of each solution, add
System suitability— separately the mobile phase to make exactly 50 mL, and use
Test for required detectability: To exactly 2 mL of the these solutions as the sample solution and the standard solu-
standard solution add a mixture of acetonitrile for liquid tion, respectively. Perform the test with exactly 10 mL each
chromatography and the mobile phase A (3:2) to make ex- of the sample solution and standard solution as directed
actly 20 mL. Confirm that the peak area of clopidogrel ob- under Liquid Chromatography <2.01> according to the fol-
tained with 10 mL of this solution is equivalent to 7 to 13z lowing conditions, and determine the peak areas, AT and AS,
of that obtained with 10 mL of the standard solution. of clopidogrel in each solution.
mL of the standard solution under the above operating con- Amount (mg) of clopidogrel sulfate (C16H16ClNO2S.H2SO4)
ditions, the number of theoretical plates and the symmetry ＝ M S × AT / AS
factor of the peak of clopidogrel are not less than 60,000 and MS: Amount (mg) of Clopidogrel Sulfate RS taken, calcu-
not more than 2.0, respectively. lated on the anhydrous basis
with 10 mL of the standard solution under the above operat- Operating conditions—
ing conditions, the relative standard deviation of the peak Detector: An ultraviolet absorption photometer (wave-
area of clopidogrel is not more than 2.0z. length: 220 nm).
(3) Optical isomer—Dissolve 0.10 g of Clopidogrel Sul- Column: A stainless steel column 3.9 mm in inside diame-
fate in 25 mL of ethanol (99.5) for liquid chromatography, ter and 15 cm in length, packed with octadecylsilanized silica
add heptane for liquid chromatography to make 50 mL, and gel for liquid chromatography (5 mm in particle diameter).
use this solution as the sample solution. Pipet 2.5 mL of the Column temperature: A constant temperature of about
sample solution, and add a mixture of ethanol (99.5) for liq- 309C.
uid chromatography and heptane for liquid chromatography Mobile phase: Dissolve 0.87 g of sodium 1-pen-
(1:1) to make exactly 50 mL. Pipet 5 mL of this solution, tanesufonate in 1000 mL of water, and adjust to pH 2.5 with
add a mixture of ethanol (99.5) for liquid chromatography phosphoric acid. To 950 mL of this solution add 50 mL of
and heptane for liquid chromatography (1:1) to make exactly methanol. To 600 mL of this solution, add 400 mL of a mix-
50 mL, and use this solution as the standard solution. Per- ture of acetonitrile for liquid chromatography and methanol
form the test with exactly 10 mL each of the sample solution (19:1).
and standard solution as directed under Liquid Chromatog- Flow rate: Adjust so that the retention time of clopidogrel
raphy <2.01> according to the following conditions, and de- is about 8 minutes.
738 Clopidogrel Sulfate Tablets / Official Monographs JP XVII
System suitability— Operating conditions—
System performance: When the procedure is run with 10 Detector: An ultraviolet absorption photometer (wave-
mL of the standard solution under the above operating con- length: 220 nm).
ditions, the number of theoretical plates and the symmetry Column: A stainless steel column of 4.6 mm in inside
factor of the peak of clopidogrel are not less than 4500 and diameter and 15 cm in length, packed with ovomucoid-
not more than 2.0, respectively. chemically bonded amino silica gel for liquid chromatogra-
System repeatability: When the test is repeated 6 times phy (5 mm in particle diameter).
with 10 mL of the standard solution under the above operat- Column temperature: A constant temperature of about
ing conditions, the relative standard deviation of the peak 259C.
area of clopidogrel is not more than 1.0z. Mobile phase: Dissolve 1.36 g of potassium dihydrogen
phosphate in 1000 mL of water, and to 750 mL of this solu-
tion add 250 mL of acetonitrile for liquid chromatography.
Flow rate: Adjust so that the retention time of clopidogrel
is about 6 minutes.
Clopidogrel Sulfate Tablets retention time of clopidogrel, beginning after the solvent
peak.
クロピドグレル硫酸塩錠
Clopidogrel Sulfate Tablets contain not less than standard solution add the mobile phase to make exactly 100
95.0z and not more than 105.0z of the labeled mL. Confirm that the peak area of clopidogrel obtained with
amount of clopidogrel (C16H16ClNO2S: 321.82). 10 mL of this solution is equivalent to 3.5 to 6.5z of that ob-
with Clopidogrel Sulfate.
Identification To a quantity of powdered Clopidogrel ditions, the number of theoretical plates and the symmetry
Sulfate Tablets, equivalent to 75 mg of clopidogrel factor of the peak of clopidogrel are not less than 2500 and
(C16H16ClNO2S), add 50 mL of methanol, and after treating not more than 2.0, respectively.
with ultrasonic waves with occasional shaking, add methanol System repeatability: When the test is repeated 6 times
to make 100 mL. To 10 mL of this solution add methanol to with 10 mL of the standard solution under the above operat-
make 30 mL, and filter. Determine the absorption spectrum ing conditions, the relative standard deviation of the peak
of the filtrate as directed under Ultraviolet-visible Spectro- area of clopidogrel is not more than 2.0z.
Purity Related substances—Keep the sample solution and lowing method: it meets the requirement.
the standard solution at 59C or below and use within 24 To 1 tablet of Clopidogrel Sulfate Tablets add a suitable
hours. Take a quantity of Clopidogrel Sulfate Tablets amount of the mobile phase, treat with ultrasonic waves with
equivalent to 0.15 g of clopidogrel (C16H16ClNO2S), add 120 occasional shaking until the tablet is disintegrated, and add
mL of the mobile phase, treat with ultrasonic waves with the mobile phase to make exactly 50 mL. Filter the solution
occasional shaking until the tablets are disintegrated, and through a membrane filter with a pore size not exceeding
add the mobile phase to make 200 mL. Centrifuge this solu- 0.45 mm. Discard the first 10 mL of the filtrate, pipet 2 mL
tion, to 10 mL of the supernatant liquid add the mobile of the subsequent filtrate, add exactly V/5 mL of the inter-
phase to make 30 mL, and filter through a membrane filter nal standard solution, and add the mobile phase to make
with a pore size not exceeding 0.45 mm. Discard the first 10 V mL so that each mL contains about 0.1 mg of clopidogrel
mL of the filtrate, and use the subsequent filtrate as the sam- (C16H16ClNO2S). Use this solution as the sample solution.
ple solution. Pipet 2 mL of the sample solution, add the mo- Then, proceed as directed in the Assay.
Amount (mg) of clopidogrel (C16H16ClNO2S)
＝ MS × QT/QS × V/10 × 0.766
under Liquid Chromatography <2.01> according to the fol- MS: Amount (mg) of Clopidogrel Sulfate RS taken, calcu-
lowing conditions, and determine each peak area by the au- lated on the anhydrous basis
tomatic integration method: the area of the peak, having the
Internal standard solution—A solution of isopropyl para-
relative retention times of about 0.3, about 0.5 and about 0.9
hydroxybenzoate in the mobile phase (1 in 1500).
to clopidogrel, obtained from the sample solution is not
larger than 3/10 times the peak area of clopidogrel obtained Dissolution <6.10> When the test is performed at 50 revolu-
from the standard solution. The area of the peak having the tions per minute according to the Paddle method, using 900
relative retention time of about 2.0 from the sample solution mL of water as the dissolution medium, the dissolution rate
is not larger than 1.2 times the peak area of clopidogrel from of 25-mg tablet in 30 minutes is not less than 70z, and that
the standard solution. The area of the peak other than of 75-mg tablet in 45 minutes is not less than 80z.
clopidogrel and the peaks mentioned above from the sample Start the test with 1 tablet of Clopidogrel Sulfate Tablets,
solution is not larger than 1/10 times the peak area of withdraw not less than 20 mL of the medium at the specified
clopidogrel from the standard solution. The total area of the minute after starting the test, and filter through a membrane
peaks other than clopidogrel from the sample solution is not filter with a pore size not exceeding 0.45 mm. Discard the
larger than 1.7 times the peak area of clopidogrel from the first 10 mL of the filtrate, pipet V mL of the subsequent fil-
standard solution. trate, add water to make exactly V? mL so that each mL con-
tains about 28 mg of clopidogrel (C16H16ClNO2S), and use
JP XVII Official Monographs / Clorazepate Dipotassium 739
this solution as the sample solution. Separately, weigh accu- mL of the standard solution under the above operating con-
rately about 30 mg of Clopidogrel Sulfate RS (separately de- ditions, the internal standard and clopidogrel are eluted in
termine the water <2.48> in the same manner as Clopidogrel this order with the resolution between these peaks being not
Sulfate), dissolve in 5 mL of methanol, and add water to less than 4.
make exactly 100 mL. Pipet 6 mL of this solution, add water System repeatability: When the test is repeated 6 times
to make exactly 50 mL, and use this solution as the standard with 10 mL of the standard solution under the above operat-
solution. Determine the absorbances, AT and AS, at 240 nm ing conditions, the relative standard deviation of the ratio of
of the sample solution and standard solution as directed the peak area of clopidogrel to that of the internal standard
under Ultraviolet-visible Spectrophotometry <2.24>, using is not more than 1.0z.
water as a blank.
of clopidogrel (C16H16ClNO2S)
＝ MS × AT/AS × V?/V × 1/C × 108 × 0.766 Clorazepate Dipotassium
MS: Amount (mg) of Clopidogrel Sulfate RS taken, calcu-
クロラゼプ酸二カリウム
C: Labeled amount (mg) of clopidogrel (C16H16ClNO2S) in
1 tablet
Assay To 20 tablets of Clopidogrel Sulfate Tablets add 400
mL of the mobile phase, treat with ultrasonic waves with oc-
casional shaking until the tablets are disintegrated, add the
mobile phase to make exactly 500 mL, and filter through a
C16H10ClKN2O3.KOH: 408.92
Discard the first 10 mL of the filtrate, pipet 5 mL of the
Monopotassium 7-chloro-2-oxo-5-phenyl-2,3-dihydro-
subsequent filtrate, add the mobile phase to make exactly
1H-1,4-benzodiazepine-3-carboxylate
V mL so that each mL contains about 0.5 mg of clopidogrel
mono (potassium hydroxide)
(C16H16ClNO2S). Pipet 4 mL of this solution, add exactly 4
[57109-90-7]
mL of the internal standard solution and the mobile phase to
Clorazepate Dipotassium, when dried, contains not
Separately, weigh accurately about 33 mg of Clopidogrel
less than 98.5z and not more than 101.0z of cloraze-
Sulfate RS (separately determine the water <2.48> in the same
pate dipotassium (C16H10ClKN2O3.KOH).
manner as Clopidogrel Sulfate), and dissolve in the mobile
phase to make exactly 50 mL. Pipet 4 mL of this solution, Description Clorazepate Dipotassium occurs as white to
add exactly 4 mL of the internal standard solution and the light yellow, crystals or crystalline powder.
mobile phase to make 20 mL, and use this solution as the It is freely soluble in water, and very slightly soluble in
standard solution. Perform the test with 10 mL each of the ethanol (99.5).
sample solution and standard solution as directed under Liq- It dissolves in acetic acid (100).
uid Chromatography <2.01> according to the following con- The pH of a solution obtained by dissolving 1 g of
ditions, and calculate the ratios, QT and QS, of the peak area Clorazepate Dipotassium in 100 mL of water is between 11.5
of clopidogrel to that of the internal standard. and 12.5.
It gradually turns yellow on exposure to light.
Amount (mg) of clopidogrel (C16H16ClNO2S) in 1 tablet
of Clopidogrel Sulfate Tablets Identification (1) Carefully and gradually ignite to red-
＝ MS × QT/QS × V/10 × 0.766 ness 30 mg of Clorazepate Dipotassium with 50 mg of so-
dium. After cooling, add 3 drops of ethanol (99.5) and 5 mL
MS: Amount (mg) of Clopidogrel Sulfate RS taken, calcu-
of water, mix well, and filter: the filtrate responds to the
Qualitative Tests <1.09> for chloride.
Internal standard solution—A solution of isopropyl para- (2) Determine the absorption spectrum of a solution of
hydoxybenzoate in the mobile phase (1 in 1500). Clorazepate Dipotassium (1 in 200,000) as directed under Ul-
Operating conditions— traviolet-visible Spectrophotometry <2.24>, and compare the
Detector: An ultraviolet absorption photometer (wave- spectrum with the Reference Spectrum: both spectra exhibit
length: 220 nm). similar intensities of absorption at the same wavelengths.
Column: A stainless steel column of 3.9 mm in inside di- (3) Determine the infrared absorption spectrum of
ameter and 15 cm in length, packed with octadecylsilanized Clorazepate Dipotassium as directed in the potassium bro-
silica gel for liquid chromatography (5 mm in particle diame- mide disk method under Infrared Spectrophotometry <2.25>,
ter). and compare the spectrum with the Reference Spectrum:
Column temperature: A constant temperature of about both spectra exhibit similar intensities of absorption at the
309 C. same wave numbers.
Mobile phase: Dissolve 0.87 g of sodium 1-pentanesul- (4) Clorazepate Dipotassium responds to the Qualitative
fonate in 1000 mL of water, and adjust to pH 2.5 with phos- Tests <1.09> (1) for potassium salt.
phoric acid. To 950 mL of this solution add 50 mL of metha-
Purity (1) Chloride <1.03>—Dissolve 1.0 g of Clorazepate
nol. To 600 mL of this solution add 400 mL of a mixture of
Dipotassium in 20 mL of water, add 20 mL of acetone, 6 mL
acetonitrile for liquid chromatography and methanol (19:1).
of dilute nitric acid and water to make 50 mL. Perform the
Flow rate: Adjust so that the retention time of clopidogrel
test with this solution as the test solution. Prepare the con-
is about 8 minutes.
trol solution as follows: To 0.40 mL of 0.01 mol/L hydro-
chloric acid VS add 20 mL of acetone, 6 mL of dilute nitric
740 Clorazepate Dipotassium Capsules / Official Monographs JP XVII
acid and water to make 50 mL (not more than 0.014z). Assay Weigh accurately about 0.15 g of Clorazepate
(2) Heavy metals <1.07>—Proceed with 1.0 g of Cloraze- Dipotassium, previously dried, dissolve in 100 mL of acetic
pate Dipotassium according to Method 2, and perform the acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid
test. Prepare the control solution with 2.0 mL of Standard VS until the color of solution changes from violet to blue-
Lead Solution (not more than 20 ppm). green through blue (indicator: 3 drops of crystal violet TS).
(3) Arsenic <1.11>—Prepare the test solution with 1.0 g Perform a blank determination in the same manner, and
of Clorazepate Dipotassium according to Method 3, and per- make any necessary correction.
(4) Related substances—Dissolve 15 mg of Clorazepate
＝ 13.63 mg of C16H10ClKN2O3.KOH
Dipotassium in 25 mL of a mixture of water, potassium car-
bonate solution (97 in 1000) and acetonitrile (3:1:1), and use Containers and storage Containers—Tight containers.
this solution as the sample solution. Pipet 1 mL of the sam- Storage—Light-resistant.
ple solution, add the mixture of water, potassium carbonate
solution (97 in 1000) and acetonitrile (3:1:1) to make exactly
200 mL, and use this solution as the standard solution. Pre- Clorazepate Dipotassium Capsules
pare these solutions quickly and perform the test within 3
minutes. Perform the test with exactly 5 mL each of the sam- クロラゼプ酸二カリウムカプセル
ple solution and standard solution as directed under Liquid
Clorazepate Dipotassium Capsules contain not
tegration method: the peak area of nordiazepam, having the
the labeled amount of clorazepate dipotassium
relative retention time of about 3.0 to clorazepic acid, ob-
(C16H10ClKN2O3.KOH: 408.92).
tained from the sample solution is not larger than the peak
area of clorazepic acid obtained from the standard solution, Method of preparation Prepare as directed under Cap-
the area of the peak other than clorazepic acid and nordia- sules, with Clorazepate Dipotassium.
zepam is not larger than 1/5 times the peak area of
Identification To 10 mL of the sample solution obtained in
clorazepic acid from the standard solution, and the total
the Assay add water to make 20 mL. Determine the absorp-
area of the peaks other than clorazepic acid is not larger than
2 times the peak area of clorazepic acid from the standard
solution. For the area of the peak of nordiazepam, multiply
the relative response factor, 0.64.
Operating conditions— Purity Related substances—Take out the contents of
Detector: An ultraviolet absorption photometer (wave- Clorazepate Dipotassium Capsules, and powder. To a por-
length: 232 nm). tion of the powder, equivalent to 15 mg of Clorazepate
Column: A stainless steel column 4.6 mm in inside diame- Dipotassium, add a mixture of water, potassium carbonate
ter and 15 cm in length, packed with octadecylsilanized silica solution (97 in 1000) and acetonitrile (3:1:1) to make 25 mL,
gel for liquid chromatography (5 mm in particle diameter). and shake for 10 minutes. Filter the solution through a mem-
Column temperature: A constant temperature of about brane filter with a pore size not exceeding 0.45 mm, discard
259 C. the first 5 mL of the filtrate, and use the subsequent filtrate
Mobile phase: Dissolve 13.8 g of sodium dihydrogen phos- as the sample solution. Pipet 1 mL of the sample solution,
phate dihydrate in 500 mL of water, and adjust to pH 8.0 add a mixture of water, potassium carbonate solution (97 in
with sodium hydroxide TS. To 100 mL of this solution add 1000) and acetonitrile (3:1:1) to make exactly 200 mL, and
400 mL of acetonitrile and 300 mL of water. use this solution as the standard solution. Then, proceed as
Flow rate: Adjust so that the retention time of clorazepic directed in the Purity (4) under Clorazepate Dipotassium:
acid is about 1.3 minutes. the peak area of nordiazepam, having the relative retention
Time span of measurement: About 10 times as long as the time of about 3.0 to clorazepic acid, obtained from the sam-
retention time of clorazepic acid, beginning after the solvent ple solution is not larger than 3 times the peak area of clora-
peak. zepic acid obtained from the standard solution, and the total
System suitability— area of the peaks other than clorazepic acid and nordiazep-
Test for required detectability: To exactly 5 mL of the am is not larger than the peak area of clorazepic acid from
standard solution add the mixture of water, potassium car- the standard solution. For the peak area of nordiazepam,
bonate solution (97 in 1000) and acetonitrile (3:1:1) to make multiply the relative response factor, 0.64.
exactly 25 mL. Confirm that the peak area of clorazepic acid
To 1 capsule of Clorazepate Dipotassium Capsules add 70
mL of water, shake for 15 minutes, and add water to make
exactly 100 mL. Centrifuge the solution, pipet V mL of the
tor of the peak of clorazepic acid are not less than 3000 and
supernatant liquid, add water to make exactly V? mL so that
each mL contains about 12 mg of clorazepate dipotassium
(C16H10ClKN2O3.KOH), and use this solution as the sample
of clorazepic acid is not more than 1.5z. Amount (mg) of clorazepate dipotassium
(C16H10ClKN2O3.KOH)
＝ MS × AT/AS × V?/V × 2/25
JP XVII Official Monographs / Clotiazepam 741
MS: Amount (mg) of clorazepate dipotassium for assay

taken Clotiazepam
クロチアゼパム
dissolution rate in 30 minutes of Clorazepate Dipotassium
Capsules is not less than 80z.
Start the test with 1 capsule of Clorazepate Dipotassium
Capsules, withdraw not less than 20 mL of the medium at
C16H15ClN2OS: 318.82
5-(2-Chlorophenyl)-7-ethyl-1-methyl-1,3-dihydro-2H-
each mL contains about 8.3 mg of clorazepate dipotassium
thieno[2,3-e][1,4]-diazepin-2-one
(C16H10ClKN2O3.KOH), and use this solution as the sample
[33671-46-4]
solution. Separately, weigh accurately about 21 mg of
clorazepate dipotassium for assay, previously dried in vacu-
Clotiazepam, when dried, contains not less than
um over phosphorus (V) oxide at 609 C for 5 hours, and dis-
98.5z of clotiazepam (C16H15ClN2OS).
solution, add water to make exactly 100 mL, and use this so- Description Clotiazepam occurs as white to light yellowish
lution as the standard solution. Determine the absorbances, white, crystals or crystalline powder. It is odorless, and has a
AT and AS, at 252 nm of the sample solution and standard slightly bitter taste.
solution as directed under Ultraviolet-visible Spectropho- It is very soluble in chloroform, freely soluble in metha-
tometry <2.24>. nol, in ethanol (95), in acetone, in acetic acid (100) and in
ethyl acetate, soluble in diethyl ether, and practically insolu-
ble in water.
of clorazepate dipotassium (C16H10ClKN2O3.KOH)
＝ MS × AT/AS × V?/V × 1/C × 36
MS: Amount (mg) of clorazepate dipotassium for assay
Identification (1) Dissolve 0.01 g of Clotiazepam in 3 mL
taken
of sulfuric acid: the solution shows a light yellow fluores-
C: Labeled amount (mg) of clorazepate dipotassium
cence under ultraviolet light (main wavelength: 365 nm).
(C16H10ClKN2O3.KOH) in 1 capsule
Assay Carefully take out the contents of not less than 20 Clotiazepam in 0.1 mol/L hydrochrolic acid TS (1 in
Clorazepate Dipotassium Capsules, weigh accurately the 100,000) as directed under Ultraviolet-visible Spectropho-
mass of the contents, and powder. Weigh accurately a por- tometry <2.24>, and compare the spectrum with the Refer-
tion of the powder, equivalent to about 15 mg of clorazepate ence Spectrum: both spectra exhibit similar intensities of ab-
dipotassium (C16H10ClKN2O3.KOH), add 70 mL of water, sorption at the same wavelengths.
shake for 15 minutes, and add water to make exactly 100 (3) Prepare the test solution with 0.01 g of Clotiazepam
mL. Centrifuge the solution, pipet 4 mL of the supernatant as directed under Oxygen Flask Combustion Method <1.06>,
liquid, add water to make exactly 50 mL, and use this solu- using 10 mL of diluted hydrogen peroxide (30) (1 in 5) as the
tion as the sample solution. Separately, weigh accurately absorbing liquid. Apply a small amount of water to the up-
about 15 mg of clorazepate dipotassium for assay, previ- per part of the Apparatus A, pull out C carefully, wash C, B
ously dried in vacuum over phosphorus (V) oxide at 609C and the inner side of A with 15 mL of methanol, and use the
for 5 hours, and dissolve in water to make exactly 100 mL. obtained solution as the test solution. Add 0.5 mL of dilute
Pipet 4 mL of this solution, add water to make exactly 50 nitric acid to 15 mL of the test solution: this solution re-
mL, and use this solution as the standard solution. Perform sponds to the Qualitative Tests <1.09> (2) for chloride. The
the test with the sample solution and standard solution as remaining test solution responds to the Qualitative Tests
directed under Ultraviolet-visible Spectrophotometry <2.24>, <1.09> (1) for sulfate.
and determine the absorbances, AT and AS, at 252 nm.
Amount (mg) of clorazepate dipotassium
(C16H10ClKN2O3.KOH)
of Clotiazepam in 10 mL of ethanol (95): the solution is clear
＝ M S × A T / AS
and is not more colored than the following control solution.
MS: Amount (mg) of clorazepate dipotassium for assay Control solution: To 5 mL of Matching Fluid C add 0.01
taken mol/L hydrochloric acid TS to make 10 mL.
(2) Chloride <1.03>—To 1.0 g of Clotiazepam add 50 mL
of water, shake for 30 minutes, and filter. To 30 mL of the
mL. Perform the test using this solution as the test solution.
Prepare the control solution with 0.25 mL of 0.01 mol/L hy-
drochloric acid VS (not more than 0.015z).
(3) Heavy metals <1.07>—Proceed with 2.0 g of Clotia-
zepam according to Method 4, and perform the test. Prepare
742 Clotrimazole / Official Monographs JP XVII
of Clotiazepam, according to Method 3, and perform the (3) Determine the infrared absorption spectrum of
test (not more than 2 ppm). Clotrimazole, previously dried, as directed in the potassium
(5) Related substances—Dissolve 0.25 g of Clotiazepam bromide disk method under Infrared Spectrophotometry
in 10 mL of acetone, and use this solution as the sample so- <2.25>, and compare the spectrum with the Reference Spec-
lution. Pipet 1 mL of the sample solution, add acetone to trum: both spectra exhibit similar intensities of absorption at
make exactly 20 mL, pipet 2 mL of this solution, add ace- the same wave numbers.
tone to make exactly 50 mL, and use this solution as the (4) Perform the test with Clotrimazole as directed under
standard solution. Perform the test with these solutions as Flame Coloration Test <1.04> (2): a green color appears.
Melting point <2.60> 142 – 1459
C
plate of silica gel with fluorescent indicator for thin-layer Purity (1) Clarity and color of solution—Dissolve 0.5 g
chromatography. Develop the plate with a mixture of chlo- of Clotrimazole in 10 mL of dichloromethane: the solution is
roform and acetone (5:1) to a distance of about 10 cm, and clear and colorless.
air-dry the plate. Examine under ultraviolet light (main (2) Chloride <1.03>—Dissolve 1.0 g of Clotrimazole in 40
wavelength: 254 nm): the spots other than the principal spot mL of N, N-dimethylformamide, add 6 mL of dilute nitric
from the sample solution are not more intense than the spot acid and water to make 50 mL. Perform the test using this
from the standard solution. solution as the test solution. Prepare the control solution
with 0.60 mL of 0.01 mol/L hydrochloric acid VS, 40 mL of
N, N-dimethylformamide, 6 mL of dilute nitric acid and
3 hours).
water to make 50 mL (not more than 0.021z).
Residue on ignition <2.44> not more than 0.1z (1 g). (3) Sulfate <1.14>—Dissolve 0.5 g of Clotrimazole in 10
mL of methanol, and add 1 mL of dilute hydrochloric acid
Assay Weigh accurately about 0.5 g of Clotiazepam, previ-
and water to make 50 mL. Perform the test using this solu-
<2.50> with 0.1 mol/L perchloric acid (potentiometric titra-
0.05 mL of 0.005 mol/L sulfuric acid VS, 10 mL of metha-
tion). Perform a blank determination in, and make any nec-
nol, 1 mL of dilute hydrochloric acid and water to make 50
essary correction.
mL (not more than 0.048z).
Each mL of 0.1 mol/L perchloric acid VS (4) Heavy metals <1.07>—Proceed with 2.0 g of
＝ 31.88 mg of C16H15ClN2OS Clotrimazole according to Method 2, and perform the test.
of Clotrimazole according to Method 3, and perform the test
Clotrimazole (6) Imidazole—Dissolve 0.10 g of Clotrimazole in ex-
actly 10 mL of dichloromethane, and use this solution as the
クロトリマゾール
sample solution. Separately, dissolve 25 mg of imidazole for
thin-layer chromatography in dichloromethane to make
exactly 50 mL. Pipet 5 mL of this solution, add dichloro-
methane to make exactly 50 mL, and use this solution as the
the plate with a mixture of methanol and chloroform (3:2) to
C22H17ClN2: 344.84
a distance of about 10 cm, and air-dry the plate. Spray
1-[(2-Chlorophenyl)(diphenyl)methyl]-1H-imidazole
evenly sodium hypochlorite TS on the plate, and air-dry the
[23593-75-1]
plate for 15 minutes, then spray evenly potassium iodide-
starch TS on the plate: the spot from the sample solution,
Clotrimazole, when dried, contains not less than
corresponding to that from the standard solution, is not
98.0z of clotrimazole (C22H17ClN2).
more intense than that from the standard solution.
Description Clotrimazole occurs as a white, crystalline (7) (2-Chlorophenyl)-diphenylmethanol—Dissolve 0.20 g
powder. It is odorless and tasteless. of Clotrimazole in exactly 10 mL of dichloromethane, and
It is freely soluble in dichloromethane and in acetic acid use this solution as the sample solution. Separately, dissolve
(100), soluble in N, N-dimethylformamide, in methanol and 10 mg of (2-chlorophenyl)-diphenylmethanol for thin-layer
in ethanol (95), slightly soluble in diethyl ether, and practi- chromatography in dichloromethane to make exactly 100
cally insoluble in water. mL, and use this solution as the standard solution. Perform
Identification (1) To 0.1 g of Clotrimazole add 10 mL of
5 mol/L hydrochloric acid TS, dissolve by heating, and cool.
tion and standard solution on a plate of silica gel with fluo-
To this solution add 3 drops of Reinecke salt TS: a light red
rescent indicator for thin-layer chromatography. Develop
the plate with a mixture of ethyl acetate and ammonia solu-
tion (28) (50:1) to a distance of about 10 cm, and air-dry the
Clotrimazole in methanol (1 in 5000) as directed under Ultra-
nm): the spot from the sample solution, corresponding to
that from the standard solution, is not more intense than
that from the standard solution.
JP XVII Official Monographs / Cloxacillin Sodium Hydrate 743
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, tained by dissolving 1.0 g of Cloxacillin Sodium Hydrate in
2 hours). 10 mL of water is clear, and its absorbance at 430 nm, deter-
mined as directed under Ultraviolet-visible Spectrophotome-
try <2.24>, is not more than 0.04.
Assay Weigh accurately about 0.35 g of Clotrimazole, pre- (2) Heavy metals <1.07>—Proceed with 1.0 g of Cloxacil-
viously dried, and dissolve in 80 mL of acetic acid (100), and lin Sodium Hydrate according to Method 2, and perform the
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio- test. Prepare the control solution with 2.0 mL of Standard
metric titration). Perform a blank determination, and make Lead Solution (not more than 20 ppm).
any necessary correction. (3) Arsenic <1.11>—Prepare the test solution with 1.0 g
of Cloxacillin Sodium Hydrate according to Method 5, and
＝ 34.48 mg of C22H17ClN2
(4) Related substances—Dissolve 50 mg of Cloxacillin
Containers and storage Containers—Well-closed contain- Sodium Hydrate in 50 mL of the mobile phase, and use this
ers. solution as the sample solution. Pipet 1 mL of the sample so-
Storage—Light-resistant. lution, add the mobile phase to make exactly 100 mL, and
Cloxacillin Sodium Hydrate solution as directed under Liquid Chromatography <2.01>
クロキサシリンナトリウム水和物 peak area by the automatic integration method: the area of
the peak other than cloxacillin obtained from the sample so-
lution is not larger than the peak area of cloxacillin obtained
other than chloxacillin from the sample solution is not larger
than 3 times the peak area of cloxacillin from the standard
solution.
C19H17ClN3NaO5S.H2O: 475.88
Monosodium (2S,5R,6R)-6-{[3-(2-chlorophenyl)-5-
methylisoxazole-4-carbonyl]amino}-3,3-dimethyl-7-oxo-4-
the Assay.
thia-1-azabicyclo[3.2.0]heptane-2-carboxylate
monohydrate
retention time of cloxacillin.
[7081-44-9]
Cloxacillin Sodium Hydrate contains not less than
Confirm that the peak area of cloxacillin obtained with 10
mL of this solution is equivalent to 7 to 13z of that obtained
tency of Cloxacillin Sodium Hydrate is expressed as
mass (potency) of cloxacillin (C19H18ClN3O5S: 435.88).
System performance: Dissolve about 50 mg of Cloxacillin
Description Cloxacillin Sodium Hydrate occurs as white to Sodium RS in a suitable amount of the mobile phase, add
light yellowish white, crystals or crystalline powder. 5 mL of a solution of guaifenesin in the mobile phase (1 in
It is freely soluble in water, in N, N-dimethylformamide 200), then add the mobile phase to make 50 mL, and use this
and in methanol, and sparingly soluble in ethanol (95). solution as the solution for system suitability test. When the
procedure is run with 10 mL of the solution for system suita-
bility test under the above operating conditions, guaifenesin
solution of Cloxacillin Sodium Hydrate in methanol (1 in
and cloxacillin are eluted in this order with the resolution be-
2500) as directed under Ultraviolet-visible Spectrophotome-
Spectrum or the spectrum of a solution of Cloxacillin So-
dium RS prepared in the same manner as the sample solu-
tion of the peak area of cloxacillin is not more than 1.0z.
(2) Determine the infrared absorption spectrum of Clox- Water <2.48> 3.0 – 4.5z (0.2 g, volumetric titration, direct
acillin Sodium Hydrate as directed in the potassium bromide titration).
Assay Weigh accurately an amount of Cloxacillin Sodium
Hydrate and Cloxacillin Sodium RS, equivalent to about 50
spectrum of Cloxacillin Sodium RS: both spectra exhibit
mg (potency), dissolve each in a suitable amount of the mo-
bile phase, add exactly 5 mL of the internal standard solu-
(3) Cloxacillin Sodium Hydrate responds to the Qualita-
tion, then add the mobile phase to make 50 mL, and use
these solutions as the sample solution and the standard solu-
D : ＋163 – ＋1719(1 g calculated tion, respectively. Perform the test with 10 mL each of the
on the anhydrous basis, water, 100 mL, 100 mm). sample solution and standard solution as directed under Liq-
pH <2.54> Dissolve 1.0 g of Cloxacillin Sodium Hydrate in
10 mL of water: the pH of the solution is between 5.0 and
of cloxacillin to that of the internal standard.
7.5.
744 Cloxazolam / Official Monographs JP XVII
Amount [ mg (potency)] of cloxacillin (C19H18ClN3O5S) green fluorescence under ultraviolet light (main wavelength:
＝ MS × QT/QS × 1000 365 nm). Add 1 mL of sodium hydroxide TS to this solution:
the color and fluorescence of this solution disappear immedi-
MS: Amount [mg (potency)] of Cloxacillin Sodium RS
ately.
taken
(2) Dissolve 0.01 g of Cloxazolam in 5 mL of dilute hy-
Internal standard solution—A solution of guaifenesin in the drochloric acid by heating in a water bath for 10 minutes.
mobile phase (1 in 200). After cooling, 1 mL of this solution responds to the Qualita-
Operating conditions— tive Tests <1.09> for primary aromatic amines.
Detector: An ultraviolet absorption photometer (wave- (3) Place 2 g of Cloxazolam in a 200-mL flask, add 50
length: 230 nm). mL of ethanol (95) and 25 mL of sodium hydroxide TS, and
Column: A stainless steel column 6 mm in inside diameter boil under a reflux condenser for 4 hours. After cooling,
and 15 cm in length, packed with octadecylsilanized silica gel neutralize with dilute hydrochloric acid, and extract with 30
for liquid chromatography (5 mm in particle diameter). mL of dichloromethane. Dehydrate with 3 g of anhydrous
Column temperature: A constant temperature of about sodium sulfate, filter, and evaporate the dichloromethane of
259 C. the filtrate. Dissolve the residue in 5 mL of methanol by
Mobile phase: Dissolve 4.95 g of diammonium hydrogen heating on a water bath, and cool immediately in an ice
phosphate in 700 mL of water, add 250 mL of acetonitrile, bath. Collect the crystals, and dry the crystals is vacuum at
adjust to pH 4.0 with phosphoric acid, and add water to 609C for 1 hour: it melts <2.60> between 879C and 919C.
make 1000 mL. (4) Determine the absorption spectrum of a solution of
Flow rate: Adjust so that the retention time of cloxacillin Cloxazolam in ethanol (99.5) (1 in 100,000) as directed under
is about 24 minutes. Ultraviolet-visible Spectrophotometry <2.24>, and compare
System suitability— the spectrum with the Reference Spectrum: both spectra
System performance: When the procedure is run with 10 exhibit similar intensities of absorption at the same wave-
mL of the standard solution under the above operating con- lengths.
ditions, guaifenesin and cloxacillin are eluted in this order (5) Proceed with Cloxazolam as directed under Flame
with the resolution between these peaks being not less than Coloration Test <1.04> (2), and perform the test: a green
25. color appears.
1 mg, ethanol (99.5), 100 mL).
the peak area of cloxacillin to that of the internal standard is Purity (1) Chloride <1.03>—To 1.0 g of Cloxazolam add
not more than 1.0z. 50 mL of water, allow to stand for 1 hour with occasional
shaking, and filter. To 25 mL of this filtrate add 6 mL of
dilute nitric acid and water to make 50 mL, and perform the
trol solution with 0.20 mL of 0.01 mol/L hydrochloric acid
Cloxazolam VS (not more than 0.014z).
(2) Heavy metals <1.07>—Proceed with 1.0 g of Clox-
クロキサゾラム
azolam according to Method 2, and perform the test. Pre-
(3) Arsenic <1.11>—Place 1.0 g of Cloxazolam in a Kjel-
dahl flask, add 5 mL of sulfuric acid and 5 mL of nitric acid,
and heat gently. Repeat the addition of 2 to 3 mL of nitric
acid at times, and continue heating until a colorless to light
yellow solution is obtained. After cooling, add 15 mL of
C17H14Cl2N2O2: 349.21
saturated ammonium oxalate solution, and heat the solution
(11bRS )-10-Chloro-11b-(2-chlorophenyl)-2,3,7,11b-
until dense white fumes are evolved, and evaporate to a
tetrahydro[1,3]oxazolo[3,2-d ][1,4]benzodiazepin-
volume of 2 to 3 mL. After cooling, dilute with water to 10
6(5H )-one
mL, and perform the test with this solution as the test solu-
[24166-13-0]
tion (not more than 2 ppm).
(4) Related substances—Dissolve 0.05 g of Cloxazolam
Cloxazolam, when dried, contains not less than
in 10 mL of dichloromethane, and use this solution as the
99.0z of cloxazolam (C17H14Cl2N2O2).
Description Cloxazolam occurs as white, crystals or crys- dichloromethane to make exactly 200 mL, and use this solu-
talline powder. It is odorless and tasteless. tion as the standard solution. Perform the test with these so-
It is freely soluble in acetic acid (100), sparingly soluble in lutions as directed under Thin-layer Chromatography <2.03>.
dichloromethane, slightly soluble in ethanol (99.5) and in Spot 10 mL each of the sample solution and standard solu-
diethyl ether, very slightly soluble in ethanol (95), and practi- tion on a plate of silica gel with fluorescent indicator for
cally insoluble in water. thin-layer chromatography. Immediately after air-drying,
It dissolves in dilute hydrochloric acid. develop the plate with a mixture of toluene and acetone (5:1)
It is gradually colored by light. to a distance of about 10 cm, and air-dry the plate. Examine
Melting point: about 2009C (with decomposition). under ultraviolet light (main wavelength: 254 nm): the spots
Identification (1) Dissolve 0.01 g of Cloxazolam in 10 mL
not more intense than that from the standard solution.
of ethanol (99.5) by heating, and add 1 drop of hydrochloric
acid: the solution shows a light yellow color and a yellow- Loss on drying <2.41> Not more than 0.5z (1 g, 1059C,
JP XVII Official Monographs / Cod Liver Oil 745
3 hours). (2) Cinnamyl cocaine—Dissolve 0.10 g of Cocaine Hy-

drochloride in 5 mL of water, and add 0.3 mL of diluted sul-
furic acid (1 in 20) and 0.10 mL of 0.02 mol/L potassium
Assay Weigh accurately about 0.5 g of Cloxazolam, previ- permanganate VS: the red color does not disappear within 30
ously dried, and dissolve in 50 mL of acetic acid (100). minutes.
Titrate <2.50> with 0.1 mol/L perchloric acid VS until the (3) Isoatropyl cocaine—Dissolve 0.10 g of Cocaine Hy-
color of the solution changes from purple through blue to drochloride in 30 mL of water in a beaker. Transfer 5 mL of
blue-green (indicator: 2 drops of crystal violet TS). Perform this solution to a test tube, add 1 drop of ammonia TS, and
a blank determination. mix. After the precipitate is coagulated, add 10 mL of water,
and transfer the mixture to the former beaker, to which 30
mL of water has been added previously. Wash the test tube
＝ 34.92 mg of C17H14Cl2N2O2
with 10 mL of water, combine the washings with the mixture
Containers and storage Containers—Tight containers. in the beaker, add 3 drops of ammonia TS to the combined
Storage—Light-resistant. mixture, and mix gently: a crystalline precipitate is pro-
duced. Allow to stand for 1 hour: the supernatant liquid is
clear.
Cocaine Hydrochloride Loss on drying <2.41> Not more than 1.0z (1 g, 1059C,
4 hours).
コカイン塩酸塩
Assay Weigh accurately about 0.5 g of Cocaine Hydro-
acetic anhydride and acetic acid (100) (7:3), and titrate <2.50>
with 0.1 mol/L perchloric acid VS (potentiometric titration).
correction.
C17H21NO4.HCl: 339.81
(1R,2R,3S,5S )-2-Methoxycarbonyl-8-methyl-8- Each mL of 0.1 mol/L perchloric acid VS
azabicyclo[3.2.1]oct-3-yl benzoate monohydrochloride ＝ 33.98 mg of C17H21NO4.HCl
[53-21-4]
Cocaine Hydrochloride, when dried, contains not
less than 98.0z of cocaine hydrochloride (C17H21NO4.
HCl).
Cod Liver Oil
Description Cocaine Hydrochloride occurs as colorless
crystals or a white crystalline powder. 肝油
It is very soluble in water, freely soluble in ethanol (95)
and in acetic acid (100), slightly soluble in acetic anhydride,
Cod Liver Oil is the fatty oils obtained from fresh
livers and pyloric appendages of Gadus macrocephalus
Identification (1) Determine the absorption spectrum of a Tilesius or Theragra chalcogramma Pallas (Gadidae).
solution of Cocaine Hydrochloride in 0.01 mol/L hydro- Cod Liver Oil contains not less than 2000 Vitamin A
chloric acid TS (1 in 10,000) as directed under Ultraviolet- Units and not more than 5000 Vitamin A Units per g.
Description Cod Liver Oil is a yellow to orange oily liquid.
with the Reference Spectrum 1: both spectra exhibit similar
It has a characteristic, slightly fishy odor and a mild taste.
intensities of absorption at the same wavelengths. Sepa-
It is miscible with chloroform.
rately, determine the absorption spectrum of a solution of
It is slightly soluble in ethanol (95), and practically insolu-
Cocaine Hydrochloride in 0.01 mol/L hydrochloric acid TS
ble in water.
It is decomposed by air or by light.
erence Spectrum 2: both spectra exhibit similar intensities of Identification Dissolve 0.1 g of Cod Liver Oil in 10 mL of
absorption at the same wavelengths. chloroform, and to 1 mL of this solution add 3 mL of an-
(2) Determine the infrared absorption spectrum of timony (III) chloride TS: a blue color develops immediately,
Cocaine Hydrochloride, previously dried, as directed in the but the color fades rapidly.
potassium bromide disk method under the Infrared Spectro-
20: 0.918 – 0.928
erence Spectrum: both spectra exhibit similar intensities of Acid value <1.13> Not more than 1.7.
Saponification value <1.13> 180 – 192
(3) A solution of Cocaine Hydrochloride (1 in 50) re-
sponds to the Qualitative Tests <1.09> (2) for chloride. Unsaponifiable matter <1.13> Not more than 3.0z.
－70 – －739 (after drying,
D: Iodine value <1.13> 130 – 170
0.5 g, water, 20 mL, 100 mm).
Purity Rancidity—No unpleasant odor of rancid oil is per-
Purity (1) Acidity—Dissolve 0.5 g of Cocaine Hydrochlo- ceptible on warming Cod Liver Oil.
ride in 10 mL of water, add 1 drop of methyl red TS, and
Assay Proceed with about 0.5 g of Cod Liver Oil, accu-
neutralize with 0.01 mol/L sodium hydroxide VS: the con-
rately weighed, as directed in Method 2 under the Vitamin A
sumed volume is not more than 1.0 mL.
746 Codeine Phosphate Hydrate / Official Monographs JP XVII
Determination <2.55>, and perform the test. solutions as directed under Thin-layer Chromatography
Storage—Light-resistant, and almost well-filled, or under
nitrogen atmosphere.
of ethanol (99.5), toluene, acetone and ammonia solution
(28) (14:14:7:1) to a distance of about 10 cm, and air-dry the
Codeine Phosphate Hydrate nm): the spots other than the principal spot from the sample
コデインリン酸塩水和物
ard solution.
titration).
Assay Dissolve about 0.5 g of Codeine Phosphate Hydrate,
accurately weighed, in 70 mL of acetic acid (100), and titrate
the solution changes from purple through blue to greenish
blue (indicator: 3 drops of crystal violet TS). Perform a
C18H21NO3.H3PO4. 1/2 H2O: 406.37
(5R,6S )-4,5-Epoxy-3-methoxy-17-methyl-7,8-
didehydromorphinan-6-ol monophosphate hemihydrate Each mL of 0.1 mol/L perchloric acid VS
[41444-62-6] ＝ 39.74 mg of C18H21NO3.H3PO4
Codeine Phosphate Hydrate contains not less
than 98.0z of codeine phosphate (C18H21NO3.H3PO4:
Description Codeine Phosphate Hydrate occurs as white to 1 Codeine Phosphate Powder
yellowish white, crystals or crystalline powder.
It is freely soluble in water and in acetic acid (100), slightly コデインリン酸塩散 1
soluble in methanol and in ethanol (95), and practically in-
soluble in diethyl ether.
1z Codeine Phosphate Powder contains not less
The pH of a solution of 1.0 g of Codeine Phosphate Hy-
than 0.90z and not more than 1.10z of codeine
drate in 10 mL of water is between 3.0 and 5.0.
phosphate hydrate (C18H21NO3.H3PO4. 1/2 H2O:
It is affected by light.
406.37).
solution of Codeine Phosphate Hydrate (1 in 10,000) as
directed under Ultraviolet-visible Spectrophotometry <2.24>, Codeine Phosphate Hydrate 10 g
and compare the spectrum with the Reference Spectrum: Lactose Hydrate a sufficient quantity
both spectra exhibit similar intensities of absorption at the To make 1000 g
same wavelengths.
(2) Determine the infrared absorption spectrum of Prepare as directed under Granules or Powders, with the
Codeine Phosphate Hydrate, previously dried at 1059C for above ingredients.
4 hours, as directed in the potassium bromide disk method Identification Determine the absorption spectrum of a so-
under Infrared Spectrophotometry <2.25>, and compare the lution of 1z Codeine Phosphate Powder (1 in 100) as di-
spectrum with the Reference Spectrum: both spectra exhibit rected under Ultraviolet-visible Spectrophotometry <2.24>: it
similar intensities of absorption at the same wave numbers. exhibits a maximum between 283 nm and 287 nm.
(3) A solution of Codeine Phosphate Hydrate (1 in 20)
responds to the Qualitative Tests <1.09> (1) for phosphate. Dissolution <6.10> When the test is performed at 50 revolu-
－98 – －1029 (0.4 g calcu-
D: mL of water as the dissolution medium, the dissolution rate
lated on the anhydrous basis, water, 20 mL, 100 mm). in 15 minutes of 1z Codeine Phosphate Powder is not less
Purity (1) Chloride <1.03>—Perform the test with 0.5 g of than 85z.
Codeine Phosphate Hydrate. Prepare the control solution Start the test with about 2 g of 1z Codeine Phosphate
with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more Powder, accurately weighed, withdraw not less than 20 mL
than 0.021z). of the medium at the specified minute after starting the test,
(2) Sulfate <1.14>—Perform the test with 0.20 g of and filter through a membrane filter with a pore size not
Codeine Phosphate Hydrate. Prepare the control solution exceeding 0.45 mm. Discard the first 10 mL of the filtrate,
with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than and use the subsequent filtrate as the sample solution. Sepa-
0.240z). rately, weigh accurately about 28 mg of codeine phosphate
(3) Related substances—Dissolve 0.20 g of Codeine hydrate for assay (separately determine the water <2.48> in
Phosphate Hydrate in 10 mL of a mixture of 0.01 mol/L the same manner as Codeine Phosphate Hydrate), and dis-
hydrochloric acid TS and ethanol (99.5) (4:1), and use this solve in water to make exactly 100 mL. Pipet 4 mL of this
solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 50 mL, and use this so-
solution, add a mixture of 0.01 mol/L hydrochloric acid TS lution as the standard solution. Perform the test with exactly
and ethanol (99.5) (4:1) to make exactly 100 mL, and use this 50 mL each of the sample solution and standard solution as
solution as the standard solution. Perform the test with these directed under Liquid Chromatography <2.01> according to
JP XVII Official Monographs / 10 Codeine Phosphate Powder 747
the following conditions, and determine the peak areas, AT System repeatability: When the test is repeated 5 times
and AS, of codeine in each solution. with 20 mL of the standard solution under the above operat-
the peak area of codeine to that of the internal standard is
of codeine phosphate hydrate (C18H21NO3.H3PO4. 1/2 H2O)
not more than 1.0z.
＝ MS/MT × AT/AS × 36/5 × 1.023
MS: Amount (mg) of codeine phosphate hydrate for assay
MT: Amount (g) of 1z Codeine Phosphate Powder taken
10 Codeine Phosphate Powder
Proceed as directed in the operating conditions in the コデインリン酸塩散 10
Assay.
10z Codeine Phosphate Powder contains not less
than 9.3z and not more than 10.7z of codeine phos-
phate hydrate (C18H21NO3.H3PO4. 1/2 H2O: 406.37).
factor of the peak of codeine are not less than 3000 and not Method of preparation
Codeine Phosphate Hydrate 100 g
Lactose Hydrate a sufficient quantity
ing conditions, the relative standard deviation of the peak To make 1000 g
area of codeine is not more than 2.0z. Prepare as directed under Powders, with the above ingre-
Assay Weigh accurately about 5 g of 1z Codeine Phos- dients.
phate Powder, dissolve in water to make exactly 100 mL, Identification Determine the absorption spectrum of a so-
then pipet 10 mL of this solution, add exactly 10 mL of the lution of 10z Codeine Phosphate Powder (1 in 1000) as di-
internal standard solution, and use this solution as the sam- rected under Ultraviolet-visible Spectrophotometry <2.24>: it
ple solution. Separately, weigh accurately about 50 mg of exhibits a maximum between 283 nm and 287 nm.
codeine phosphate hydrate for assay (previously determine
the water <2.48> in the same manner as Codeine Phosphate Dissolution <6.10> When the test is performed at 50 revolu-
Hydrate), dissolve in water to make exactly 100 mL, then tions per minute according to the Paddle method, using 900
pipet 10 mL of this solution, add exactly 10 mL of the inter- mL of water as the dissolution medium, the dissolution rate
nal standard solution, and use this solution as the standard in 15 minutes of 10z Codeine Phosphate Powder is not less
solution. Perform the test with 20 mL each of the sample so- than 85z.
lution and standard solution as directed under Liquid Chro- Start the test with about 0.2 g of 10z Codeine Phosphate
matography <2.01> according to the following conditions, Powder, accurately weighed, withdraw not less than 20 mL
and calculate the ratios, QT and QS, of the peak area of of the medium at the specified minute after starting the test,
codeine to that of the internal standard. and filter through a membrane filter with a pore size not
Amount (mg) of codeine phosphate hydrate and use the subsequent filtrate as the sample solution. Sepa-
(C18H21NO3.H3PO4. 1/2 H2O) rately, weigh accurately about 28 mg of codeine phosphate
＝ MS × QT/QS × 1.023 hydrate for assay (separately determine the water <2.48> in
MS: Amount (mg) of codeine phosphate hydrate for assay the same manner as Codeine Phosphate Hydrate), and dis-
taken, calculated on the anhydrous basis solve in water to make exactly 100 mL. Pipet 4 mL of this
Internal standard solution—A solution of etilefrine hydro- lution as the standard solution. Perform the test with exactly
chloride (3 in 10,000). 50 mL each of the sample solution and standard solution as
Operating conditions— directed under Liquid Chromatography <2.01> according to
Detector: An ultraviolet absorption photometer (wave- the following conditions, and determine the peak areas, AT
length: 280 nm). and AS, of codeine in each solution.
ter and 15 cm in length, packed with octadecylsilanized silica Dissolution rate (z) with respect to the labeled amount
gel for liquid chromatography (5 mm in particle diameter). of codeine phosphate hydrate (C18H21NO3.H3PO4. 1/2 H2O)
Column temperature: A constant temperature of about ＝ MS/MT × AT/AS × 18/25 × 1.023
409 C. MS: Amount (mg) of codeine phosphate hydrate for assay
Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in taken, calculated on the anhydrous basis
500 mL of diluted phosphoric acid (1 in 1000), and adjust MT: Amount (g) of 10z Codeine Phosphate Powder
the pH to 3.0 with sodium hydroxide TS. To 240 mL of this
solution add 70 mL of tetrahydrofuran, and mix. Operating conditions—
Flow rate: Adjust so that the retention time of codeine is Proceed as directed in the operating conditions in the
about 10 minutes. Assay.
System suitability— System suitability—
System performance: When the procedure is run with 20 System performance: When the procedure is run with 50
mL of the standard solution under the above operating con- mL of the standard solution under the above operating con-
ditions, codeine and the internal standard are eluted in this ditions, the number of theoretical plates and the symmetry
order with the resolution between these peaks being not less factor of the peak of codeine are not less than 3000 and not
than 4. more than 2.0, respectively.
748 Codeine Phosphate Tablets / Official Monographs JP XVII
System repeatability: When the test is repeated 6 times with Codeine Phosphate Hydrate.
Identification To a quantity of powdered Codeine Phos-
phate Tablets, equivalent to 0.1 g of Codeine Phosphate
area of codeine is not more than 2.0z.
Hydrate, add 20 mL of water, shake, and filter. To 2 mL of
Assay Weigh accurately about 2.5 g of 10z Codeine Phos- the filtrate add water to make 100 mL, and determine the
phate Powder, dissolve in water to make exactly 100 mL, absorption spectrum as directed under Ultraviolet-visible
then pipet 2 mL of this solution, add exactly 10 mL of the in- Spectrophotometry <2.24>: it exhibits a maximum between
ternal standard solution and water to make 20 mL, and use 283 nm and 287 nm.
rately about 50 mg of codeine phosphate hydrate for assay
(previously determine the water <2.48> in the same manner as
Codeine Phosphate Hydrate), dissolve in water to make ex-
To 1 tablet of Codeine Phosphate Tablets add 3V/25 mL
actly 100 mL, then pipet 10 mL of this solution, add exactly
of water to disintegrate, add 2V/25 mL of diluted dilute sul-
10 mL of the internal standard solution, and use this solu-
furic acid (1 in 20), and treat with ultrasonic waves for 10
minutes. To this solution add exactly 2V/25 mL of the inter-
nal standard solution, add water to make V mL so that each
mL contains about 0.2 mg of codeine phosphate hydrate
(C18H21NO3.H3PO4. 1/2 H2O), filter, and use the filtrate as the
the peak area of codeine to that of the internal standard:
Amount (mg) of codeine phosphate hydrate of codeine phosphate hydrate for assay (separately, deter-
(C18H21NO3.H3PO4. 1/2 H2O) mine the water <2.48> in the same manner as Codeine Phos-
＝ MS × QT/QS × 5 × 1.023 phate Hydrate), and dissolve in water to make exactly 100
mL. Pipet 10 mL of this solution, add exactly 2 mL of the
internal standard solution, add water to make 25 mL, and
use this solution as the standard solution. Proceed as di-
Internal standard solution—A solution of etilefrine hydro- rected in the Assay.
chloride (3 in 10,000).
Amount (mg) of codeine phosphate hydrate
(C18H21NO3.H3PO4. 1/2 H2O)
＝ MS × QT/QS × V/250 × 1.023
length: 280 nm).
Column: A stainless steel column 4.6 mm in inside diame- MS: Amount (mg) of codeine phosphate hydrate for assay
ter and 15 cm in length, packed with octadecylsilanized silica taken, calculated on the anhydrous basis
Internal standard solution—A solution of ethylefurin hydro-
chloride (3 in 2000).
409 C.
Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in Dissolution <6.10> When the test is performed at 50 revolu-
500 mL of diluted phosphoric acid (1 in 1000), and adjust tions per minute according to the Paddle method, using 900
the pH to 3.0 with sodium hydroxide TS. To 240 mL of this mL of water as the dissolution medium, the dissolution rate
solution add 70 mL of tetrahydrofuran, and mix. in 30 minutes of Codeine Phosphate Tablets is not less than
Flow rate: Adjust so that the retention time of codeine is 80z.
about 10 minutes. Start the test with 1 tablet of Codeine Phosphate Tablets,
System suitability— withdraw not less than 20 mL of the medium at the specified
System performance: When the procedure is run with 20 minute after starting the test, and filter through a membrane
mL of the standard solution under the above operating con- filter with a pore size not exceeding 0.45 mm. Discard the
ditions, codeine and the internal standard are eluted in this first 10 mL of the filtrate, pipet V mL of the subsequent
order with the resolution between these peaks being not less filtrate, add water to make exactly V? mL so that each mL
than 4. contains about 5.6 mg of codeine phosphate hydrate
System repeatability: When the test is repeated 5 times (C18H21NO3.H3PO4. 1/2 H2O), and use this solution as the
with 20 mL of the standard solution under the above operat- sample solution. Separately, weigh accurately about 28 mg
ing conditions, the relative standard deviation of the ratios of codeine phosphate hydrate for assay (separately, deter-
of the peak area of codeine to that of the internal standard is mine the water <2.48> in the same manner as Codeine Phos-
not more than 1.0z. phate Hydrate), dissolve in water to make exactly 100 mL.
Codeine Phosphate Tablets <2.01> according to the following conditions, and determine
the peak areas, AT and AS, of codeine in each solution.
コデインリン酸塩錠
Dissolution rate (z) with respect to the labeled amount of
codeine phosphate hydrate (C18H21NO3.H3PO4. 1/2 H2O)
Codeine Phosphate Tablets contain not less ＝ MS × AT/AS × V?/V × 1/C × 18 × 1.023
than 93.0z and not more than 107.0z of the
labeled amount of codeine phosphate hydrate MS: Amount (mg) of codeine phosphate hydrate for assay
(C18H21NO3.H3PO4. 1/2 H2O: 406.37) taken, calculated on the anhydrous basis
C: Labeled amount (mg) of codeine phosphate hydrate
(C18H21NO3.H3PO4. 1/2 H2O) in 1 tablet
JP XVII Official Monographs / Colchicine 749
Operating conditions— Containers and storage Containers—Tight containers.

Assay.
System suitability— Colchicine
mL of the standard solution under the above operating con- コルヒチン
factor of the peak of codeine are not less than 5000 and not
area of codeine is not more than 2.0z.
C22H25NO6: 399.44
Codeine Phosphate Tablets. Weigh accurately a portion of
N-[(7S )-(1,2,3,10-Tetramethoxy-9-oxo-
the powder, equivalent to about 0.1 g of codeine phosphate
5,6,7,9-tetrahydrobenzo[a]heptalen-7-yl)]acetamide
hydrate (C18H21NO3.H3PO4. 1/2 H2O), add 30 mL of water,
[64-86-8]
shake, add 20 mL of diluted dilute sulfuric acid (1 in 20),
treat the mixture with ultrasonic waves for 10 minutes, and
Colchicine contains not less than 97.0z and not
add water to make exactly 100 mL. Filter the solution, then
more than 102.0z of colchicine (C22H25NO6), calcu-
pipet 5 mL of the filtrate, add exactly 10 mL of the internal
lated on the anhydrous and residual ethyl acetate-free
standard solution and water to make 20 mL, and use this so-
basis.
about 50 mg of codeine phosphate hydrate for assay (previ- Description Colchicine occurs as a yellowish white powder.
ously determine the water <2.48> in the same manner as It is very soluble in methanol, freely soluble in N, N-
Codeine Phosphate Hydrate), dissolve in water to make ex- dimethylformamide, in ethanol (95) and in acetic anhydride,
actly 100 mL, then pipet 10 mL of this solution, add exactly and sparingly soluble in water.
10 mL of the internal standard solution, and use this solu- It is colored by light.
solution of Colchicine in ethanol (95) (1 in 100,000) as
the peak area of codeine to that of the internal standard.
Amount (mg) of codeine phosphate hydrate same wavelengths.
(C18H21NO3.H3PO4. 1/2 H2O) (2) To 1 g of potassium bromide for infrared absorption
＝ MS × QT/QS × 2 × 1.023 spectrum add 0.5 mL of a solution of Colchicine in methanol
(1 in 50), grind thoroughly, and dry in vacuum at 809C for 1
hour. Determine the infrared absorption spectrum of this
powder as directed in the potassium bromide disk method
Internal standard solution—A solution of etilefrine hydro- under Infrared Spectrophotometry <2.25>, and compare the
chloride (3 in 10,000). spectrum with the Reference Spectrum: both spectra exhibit
Operating conditions— similar intensities of absorption at the same wave numbers.
Optical rotation <2.49> [a]20 D : －235 – －2509(0.1 g calcu-
length: 280 nm).
lated on the anhydrous basis and corrected by the amount of
ethyl acetate, ethanol (95), 10 mL, 100 mm).
gel for liquid chromatography (5 mm in particle diameter). Purity (1) Colchiceine—Dissolve 0.10 g of Colchicine in
Column temperature: A constant temperature of about 10 mL of water, and to 5 mL of this solution add 2 drops of
409 C. iron (III) chloride TS: no definite green color develops.
Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in (2) Chloroform and ethyl acetate—Weigh accurately
500 mL of diluted phosphoric acid (1 in 1000), and adjust about 0.6 g of Colchicine, dissolve in exactly 2 mL of the
the pH to 3.0 with sodium hydroxide TS. To 240 mL of this internal standard solution, add N, N-dimethylformamide to
solution add 70 mL of tetrahydrofuran, and mix. make 10 mL, and use this solution as the sample solution.
Flow rate: Adjust so that the retention time of codeine is Separately, weigh 0.30 g of chloroform using a 100-mL volu-
about 10 minutes. metric flask containing about 20 mL of N, N-dimethylfor-
System suitability— mamide, and add N, N-dimethylformamide to make exactly
System performance: When the procedure is run with 20 100 mL. Pipet 2 mL of this solution, add N, N-dimethylfor-
mL of the standard solution under the above operating con- mamide to make exactly 200 mL, and use this solution as the
ditions, codeine and the internal standard are eluted in this standard solution (1). Separately, weigh accurately about
order with the resolution between these peaks being not less 1.8 g of ethyl acetate using a 100-mL volumetric flask con-
than 4. taining about 20 mL of N, N-dimethylformamide, and add
System repeatability: When the test is repeated 5 times N, N-dimethylformamide to make exactly 100 mL. Pipet 2
with 20 mL of the standard solution under the above operat- mL of this solution, add exactly 2 mL of the internal stand-
ing conditions, the relative standard deviation of the ratios ard solution and N, N-dimethylformamide to make 10 mL,
of the peak area of codeine to that of the internal standard is and use this solution as the standard solution (2). Perform
not more than 1.0z. the test with exactly 2 mL each of the sample solution and
750 Colestimide / Official Monographs JP XVII
standard solutions (1) and (2) as directed under Gas Chro- Detector: An ultraviolet absorption photometer (wave-
matography <2.02> according to the following conditions: length: 254 nm).
the peak area of chloroform from sample solution is not Column: A stainless steel column 4.6 mm in inside diame-
larger than that from the standard solution (1). Calculate the ter and 25 cm in length, packed with octylsilanized silica gel
ratios of the peak area of ethyl acetate to that of the internal for liquid chromatography (5 mm in particle diameter).
standard, QT and QS, of the sample solution and standard Column temperature: A constant temperature of about
solution (2), and calculate the amount of ethyl acetate by the 259C.
following formula: the amount of ethyl acetate is not more Mobile phase: To 450 mL of 0.05 mol/L potassium dihy-
than 6.0z. drogen phosphate TS add methanol to make 1000 mL.
Adjust the pH to 5.5 with diluted phosphoric acid (7 in 200).
Amount (z) of ethyl acetate (C4H8O2)
Flow rate: Adjust so that the retention time of colchicine
＝ M S / M T × QT / QS × 2
is about 7 minutes.
MS: Amount (g) of ethyl acetate taken Time span of measurement: About 2 times as long as the
MT: Amount (g) of Colchicine taken retention time of colchicine, beginning after the solvent
peak.
Internal standard solution—A solution of 1-propanol in
N, N-dimethylformamide (3 in 200).
Test for required detectability: Pipet 1 mL of the sample
solution, and add diluted methanol (1 in 2) to make exactly
50 mL. Confirm that the peak area of colchicine obtained
from 20 mL of this solution is equivalent to 1.4 to 2.6z of
and 30 m in length, coated inside surface with polyethylene
that obtained from 20 mL of the sample solution.
glycol 20 M for gas chromatography 1.0 mm in thickness.
Column temperature: 609 C for 7 minutes, then up to
mL of the sample solution under the above operating condi-
1009C at a rate of 409 C per minute if necessary, and hold at
1009C for 10 minutes.
tor of the peak of colchicine are not less than 6000 and not
about 1309C.
with 20 mL of the sample solution under the above operating
2009C.
of colchicine is not more than 2.0z.
Flow rate: Adjust so that the retention time of ethyl ace-
tate is about 3 minutes. Water <2.48> Not more than 2.0z (0.5 g, volumetric titra-
Split ratio: 1:20. tion, back titration).
Assay Weigh accurately about 0.4 g of Colchicine, dissolve
in 25 mL of acetic anhydride, and titrate <2.50> with 0.05
solution (2), and add N, N-dimethylformamide to make
exactly 25 mL. Pipet 1 mL of this solution, and add N, N-
dimethylformamide to make exactly 50 mL. Confirm that
the peak area of ethyl acetate obtained from 2 mL of this so-
lution is equivalent to 0.11 to 0.21z of that obtained from Each mL of 0.05 mol/L perchloric acid VS
2 mL of the standard solution (2). ＝ 19.97 mg of C22H25NO6
System performance: To 1 mL of chloroform add N, N-
dimethylformamide to make 10 mL. To 1 mL of this solu-
tion add 2 mL of ethyl acetate and N, N-dimethylformamide
to make 100 mL. To 2 mL of this solution add 2 mL of the
internal standard solution and N, N-dimethylformamide to
make 10 mL. When the procedure is run with 2 mL of this so- Colestimide
lution under the above operating conditions, ethyl acetate,
コレスチミド
chloroform and the internal standard are eluted in this order
with the resolution between the peaks of chloroform and the [95522-45-5]
internal standard being not less than 2.0.
System repeatability: When the test is repeated 3 times Colestimide is an anion exchange resin, composed
with 2 mL of the standard solution (2) under the above oper- of a copolymer of 2-methylimidazole and 1-chloro-
ating conditions, the relative standard deviation of the ratio 2,3-epoxypropane.
of the peak area of ethyl acetate to that of the internal stand- It contains not less than 18.0z and not more than
ard is not more than 3.0z. 20.0z of chlorine (Cl: 35.45), calculated on the dried
(3) Related substances—Dissolve 60 mg of Colchicine in basis.
100 mL of diluted methanol (1 in 2). To 1 mL of this solu- Each g of Colestimide, calculated on the dried basis,
tion, add diluted methanol (1 in 2) to make 100 mL, and use exchanges with not less than 2.0 g and not more than
this solution as the sample solution. Perform the test with 20 2.4 g of cholic acid (C24H39O5: 407.56).
mL of the sample solution as directed under Liquid Chroma-
Description Colestimide occurs as a white to pale yellowish
white powder.
method. Calculate the total amount of the peaks other than
It is hygroscopic.
colchicine by the area percentage method: not more than
5.0z. Identification Determine the infrared absorption spectrum
Operating conditions— of Colestimide, previously dried, as directed in the potas-
JP XVII Official Monographs / Colestimide Granules 751
sium chloride disk method under Infrared Spectrophotome- MS: Amount (mg) of sodium cholate hydrate taken, calcu-
try <2.25>, and compare the spectrum with the Reference lated on the anhydrous basis
Spectrum: both spectra exhibit similar intensities of absorp- MT: Amount (mg) of Colestimide taken, calculated on the
tion at the same wave numbers. dried basis
Purity (1) Heavy metals <1.07>—Take 2.0 g of Coles- Internal standard solution—A solution of butyl parahy-
timide in a porcelain or platinum crucible, and carbonize by droxybenzoate in acetonitrile (1 in 80,000).
weakly heating. After cooling, add 10 mL of a solution of Operating conditions—
magnesium nitrate hexahydrate in ethanol (95) (1 in 10) and Detector: An ultraviolet absorption photometer (wave-
5 mL of hydrogen peroxide (30), and ignite the ethanol. length: 220 nm).
After cooling, add 1 mL of sulfuric acid, then, proceed ac- Column: A stainless steel column 4.6 mm in inside diame-
cording to Method 4, and perform the test. Prepare the con- ter and 25 cm in length, packed with octadecylsilanized silica
trol solution as follows: To 10 mL of a solution of magne- gel for liquid chromatography (5 mm in particle diameter).
sium nitrate hexahydrate in ethanol (95) (1 in 10) add 5 mL Column temperature: A constant temperature of about
of hydrogen peroxide (30), and ignite the ethanol. After 309C.
cooling, add 1 mL of sulfuric acid, then, proceed in the same Mobile phase: A mixture of diluted phosphoric acid (1 in
manner as for the test solution, and add 2.0 mL of Standard 1000) and acetonitrile (1:1).
Lead Solution and water to make 50 mL (not more than 10 Flow rate: Adjust so that the retention time of cholic acid
ppm). is about 7 minutes.
(2) Related substances—To exactly 0.50 g of Colestimide System suitability—
add exactly 20 mL of water, shake for 1 hour, centrifuge, System performance: When the procedure is run with 10
and use the supernatant liquid as the sample solution. Deter- mL of the standard solution under the above operating con-
mine the absorbance of the sample solution at 210 nm as di- ditions, cholic acid and the internal standard are eluted in
rected under Ultraviolet-visible Spectrophotometry <2.24>: this order with the resolution between these peaks being not
the absorbance is not more than 0.50. less than 7.
um, 1059C, 4 hours).
Residue on ignition <2.44> Not more than 0.1z (1 g). the peak area of cholic acid to that of the internal standard is
not more than 1.0z.
Degree of swelling Weigh accurately about 1 g of Coles-
timide, put in a 25-mL glass stoppered measuring cylinder Containers and storage Containers—Tight containers.
(about 11 mm in inside diameter), add 23 mL of water,
shake for 2 minutes, and add water to make 25 mL. After
standing for 2 hours, measure the volume of the resin layer, Colestimide Granules
and determine the volume per g, calculated on the dried
basis: the volume is 12 – 18 mL/g. コレスチミド顆粒
Assay (1) Chlorine—Weigh accurately about 0.2 g of
Colestimide, add 50 mL of water, and shake. Add 1 mL of Colestimide Granules contain not less than 87.0z
nitric acid and 25 mg of potassium nitrate, shake, and titrate and not more than 113.0z of the labeled amount of
<2.50> with 0.1 mol/L silver nitrate VS (potentiometric titra- colestimide.
tion). Perform a blank determination in the same manner,
ules, with Colestimide.
＝ 3.545 mg of Cl
of powdered Colestimide Granules as directed in the potas-
(2) Exchange capacity—Weigh accurately about 0.45 g sium chloride disk method under Infrared Spectrophotome-
of sodium cholate hydrate (separately determine the water), try <2.25>: it exhibits absorption at the wave numbers of
dissolve in water to make exactly 100 mL, and use this solu- about 1587 cm－1, 1528 cm－1 and 1262 cm－1.
tion as the sodium cholate standard stock solution. Sepa-
Uniformity of dosage units <6.02> Colestimide Granules in
rately, weigh accurately about 30 mg of Colestimide, add ex-
single-dose packages meet the requirement of the Mass varia-
actly 30 mL of the sodium cholate standard stock solution,
tion test.
shake for 1 hour, and centrifuge or filter through a mem-
brane filter with a pore size not exceeding 0.8 mm. Pipet 5 Disintegration <6.09> Carry out the test for 10 minutes with
mL of the supernatant liquid or the filtrate, add exactly 5 0.09 – 0.11 g of Colestimide Granules in six glass tubes of the
mL of the internal standard solution, and use this solution as apparatus: it meets the requirement.
the sample solution. Separately, pipet 5 mL of the sodium
Assay Weigh accurately about 4.5 g of sodium cholate
cholate standard stock solution, add exactly 5 mL of the in-
hydrate (separately determine the water), dissolve in water to
ternal standard solution, and use this solution as the stand-
make exactly 1000 mL, and use this solution as the sodium
cholate standard stock solution. Take out the contents of not
less than 20 single-dose packages of Colestimide Granules,
weigh accurately an amount of the contents, equivalent to
tions, and calculate the ratios, QT and QS, of the peak area
about 0.2 g of colestimide, add exactly 200 mL of the
of cholic acid to that of the internal standard.
sodium cholate standard stock solution, shake for 1 hour,
Exchanged amount (g) of cholic acid per g of Colestimide, and centrifuge. Pipet 5 mL of the supernatant liquid, add
calculated on the dried basis exactly 5 mL of the internal standard solution, and use this
＝ MS/MT × (QS － QT)/QS × 3/10 × 0.947 solution as the sample solution. Then, proceed as directed in
752 Colestimide Tablets / Official Monographs JP XVII
the Assay (2) under Colestimide. Column: A stainless steel column 4.6 mm in inside diame-
Amount (mg) of colestimide
＝ MS × (QS － QT)/QS × 1/5 × 1/2.2 × 0.947
MS: Amount (mg) of sodium cholate hydrate taken, calcu- 309C.
lated on the anhydrous basis Mobile phase: A mixture of diluted phosphoric acid (1 in
2.2: Quantity (g) of the cholic acid exchange per mg of 1000) and acetonitrile (1:1).
colestimide Flow rate: Adjust so that the retention time of cholic acid
is about 7 minutes.
Containers and storage Containers—Tight containers. mL of the standard solution under the above operating con-
ditions, cholic acid and the internal standard are eluted in
Colestimide Tablets less than 7.
コレスチミド錠 with 10 mL of the standard solution under the above operat-
the peak area of cholic acid to that of the internal standard is
Colestimide Tablets contain not less than 87.0z and
not more than 1.0z.
not more than 113.0z of the labeled amount of coles-
timide. Containers and storage Containers—Tight containers.
with Colestimide.
Colistin Sodium Methanesulfonate
Identification Powder Colestimide Tablets. Determine the
infrared absorption spectrum of a portion of the powder as コリスチンメタンスルホン酸ナトリウム
directed in the potassium chloride disk method under In-
frared Spectrophotometry <2.25>: it exhibits absorption at
the wave numbers of about 1587 cm－1, 1528 cm－1, 1262
cm－1, 1102 cm－1 and 1035 cm－1.
Disintegration <6.09> When carry out the test for 10
minutes, it meets the requirement.
Assay Weigh accurately about 0.45 g of sodium cholate hy-
drate (separately determine the water), dissolve in water to
[8068-28-8, Colistin Sodium Methanesulfonate]
make exactly 100 mL, and use this solution as the sodium
cholate standard stock solution. Separately, weigh accurately
Colistin Sodium Methanesulfonate is the sodium
the mass of not less than 20 Colestimide Tablets, and pow-
salt of colistin derivatives.
der. Weigh accurately a portion of the powder, equivalent to
It is a mixture of colistin A sodium methanesul-
about 30 mg of colestimide, add exactly 30 mL of the so-
fonate and colistin B sodium methanesulfonate.
dium cholate standard stock solution, shake for 1 hour, and
It, when dried, contains not less than 11,500 Units
centrifuge. Pipet 5 mL of the supernatant liquid, add exactly
per mg. The unit of Colistin Sodium Methanesul-
5 mL of the internal standard solution, and use this solution
fonate is expressed as mass of colistin A
as the sample solution. Separately, pipet 5 mL of the sodium
(R ＝ 6-methyloctanic acid, R? ＝ H; C53H100N16O13:
cholate standard stock solution, add exactly 5 mL of the in-
1169.46).
ternal standard solution, and use this solution as the stand-
ard solution. Perform the test with 10 mL each of the sample Description Colistin Sodium Methanesulfonate occurs as a
solution and standard solution as directed under Liquid white to light yellowish white powder.
Chromatography <2.01> according to the following condi- It is freely soluble in water, and practically insoluble in
tions, and calculate the ratios, QT and QS, of the peak area ethanol (95).
of cholic acid to that of the internal standard.
Identification (1) Dissolve 20 mg of Colistin Sodium
Amount (mg) of colestimide Methanesulfonate in 2 mL of water, add 0.5 mL of sodium
＝ MS × (QS － QT)/QS × 3/10 × 1/2.2 × 0.947 hydroxide TS, and add 5 drops of copper (II) sulfate TS
while shaking: a blue-purple color develops.
MS: Amount (mg) of sodium cholate hydrate taken, calcu-
(2) Dissolve 40 mg of Colistin Sodium Methanesulfonate
in 1 mL of 1 mol/L hydrochloric acid TS, and add 0.5 mL of
2.2: Exchanged amount (g) of cholic acid per g of coles-
dilute iodine TS: the color of iodine disappears.
timide, calculated on the dried basis
Internal standard solution—A solution of butyl parahy- Colistin Sodium Methanesulfonate, previously dried, as di-
droxybenzoate in acetonitrile (1 in 80,000). rected in the potassium bromide disk method under Infrared
Operating conditions— Spectrophotometry <2.25>, and compare the spectrum with
Detector: An ultraviolet absorption photometer (wave- the Reference Spectrum or the spectrum of dried Colistin So-
length: 220 nm). dium Methanesulfonate RS: both spectra exhibit similar in-
JP XVII Official Monographs / Colistin Sulfate 753

(4) Colistin Sodium Methanesulfonate responds to the Colistin Sulfate
Qualitative Tests <1.09> (1) for sodium salt.
コリスチン硫酸塩
pH <2.54> Dissolve 0.1 g of Colistin Sodium Methanesul-
fonate in 10 mL of water, and allow to stand for 30 minutes:
of Colistin Sodium Methanesulfonate in 10 mL of water: the
solution is clear and colorless.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Colistin
Sodium Methanesulfonate according to Method 4, and per-
Colistin A Sulfate C53H100N16O13.2 1/2 H2SO4: 1414.66
Colistin B Sulfate C52H98N16O13.2 1/2 H2SO4: 1400.63
of Colistin Sodium Methanesulfonate according to Method
[1264-72-8]
4, and perform the test (not more than 2 ppm).
(4) Free colistin—Dissolve 80 mg of Colistin Sodium
Colistin Sulfate is the sulfate of a mixture of peptide
Methanesulfonate in 3 mL of water, add 0.05 mL of a solu-
substances having antibacterial activity produced by
tion of silicotungstic acid 26-water (1 in 10), and immedi-
the growth of Bacillus polymyxa var. colistinus.
ately compare the solution with the reference suspension
It, when dried, contains not less than 16,000 units
described under Test Methods for Plastic Containers <7.02>:
per mg. The potency of Colistin Sulfate is expressed
the turbidity is not greater than that of the reference suspen-
as unit calculated from the amount of colistin A
sion (not more than 0.25z).
(C53H100N16O13: 1169.46). One unit of Colistin Sulfate
Loss on drying <2.41> Not more than 3.0z (0.1 g, reduced is equivalent to 0.04 mg of colistin A (C53H100N16O13).
C, 3 hours).
pressure, 609
Description Colistin Sulfate occurs as a white to light yel-
Assay Perform the test according to the Cylinder-plate lowish white powder.
method as directed under Microbial Assay for Antibiotics It is freely soluble in water, and practically insoluble in
<4.02> according to the following conditions. ethanol (99.5).
(i) Test organism—Escherichia coli NIHJ It is hygroscopic.
(ii) Culture medium—To 10.0 g of peptone, 30.0 g of so-
Identification (1) Dissolve 20 mg of Colistin Sulfate in 2
dium chloride, 3.0 g of meat extract and 20.0 g of agar add
mL of water, add 0.5 mL of sodium hydroxide TS, then add
1000 mL of water, then add a suitable amount of sodium hy-
5 drops of copper (II) sulfate TS while shaking: a purple
droxide TS so that the pH of the medium is being 6.5 to 6.6
color develops.
after sterilization, sterile, and use this as the seeded agar me-
(2) Dissolve 50 mg of Colistin Sulfate in 10 mL of
dium and the agar medium for base layer.
diluted hydrochloric acid (1 in 2). Transfer 1 mL of this solu-
(iii) Standard solutions—Weigh accurately an amount of
tion in a tube for hydrolysis, seal, and heat at 1359C for 5
Colistin Sodium Methanesulfonate RS, previously dried, dis-
hours. After cooling, open the tube, and evaporate the con-
solve in phosphate buffer solution (pH 6.0) to make a solu-
tent to dryness until the odor of hydrochloric acid is no more
tion containing 100,000 Units per mL, and use this solution
perceptible. Dissolve the residue in 0.5 mL of water, and use
as the standard stock solution. Keep the standard stock solu-
this solution as the sample solution. Separately, dissolve 20
tion at 109C or below and use within 7 days. Take exactly a
mg each of L-leucine, L-threonine, phenylalanine and L-ser-
suitable amount of the standard stock solution before use,
ine in 10 mL of water, and use these solutions as the stand-
and add phosphate buffer solution (pH 6.0) to make solu-
ard solution (1), (2), (3) and (4). Perform the test with these
tions so that each mL contains 10,000 Units and 2500 Units,
and use these solutions as the high concentration standard
solution and the low concentration standard solution, re-
solution (1), (2), (3) and (4) on a plate of cellulose for thin-
spectively.
(iv) Sample solutions—Weigh accurately an amount of
1-butanol, acetic acid (100), water, pyridine and ethanol
Colistin Sodium Methanesulfonate, previously dried, dis-
(99.5) (60:15:10:6:5) to a distance of about 10 cm, and dry
solve in phosphate buffer solution (pH 6.0) to make a solu-
the plate at 1059 C for 10 minutes. Spray evenly ninhydrin TS
tion containing about 100,000 Units per mL, and use this so-
on the plate, and heat at 1109C for 5 minutes: three principal
lution as the sample stock solution. Take exactly a suitable
spots are obtained from the sample solution, the R f values
amount of the sample stock solution, add phosphate buffer
of two spots of them are the same with those of the cor-
solution (pH 6.0) to make solutions so that each mL contains
responding spots obtained from the standard solution (1)
10,000 Units and 2500 Units, and use these solutions as the
and the standard solution (2), and the R f value of the rest
high concentration sample solution and the low concentra-
principal spot is about 0.1. No spot is observed at the posi-
tion sample solution, respectively.
tion corresponding to the spots obtained from the standard
Containers and storage Containers—Tight containers. solution (3) and the standard solution (4).
(3) A solution of Colistin Sulfate (1 in 20) responds to
the Qualitative Tests <1.09> (1) for sulfate.
D : －63 – －739 (1.25 g, after
drying, water, 25 mL, 100 mm).
754 Cortisone Acetate / Official Monographs JP XVII
0.10 g of Colistin Sulfate in 10 mL of water is between 4.0
and 6.0. Cortisone Acetate
Purity (1) Sulfuric acid—Weigh accurately about 0.25 g
コルチゾン酢酸エステル
of previously dried Colistin Sulfate, dissolve in a suitable
amount of water, adjust the pH to 11 with ammonia solution
(28), and add water to make 100 mL. To this solution add
exactly 10 mL of 0.1 mol/L barium chloride VS and 50 mL
of ethanol (99.5), and titrate with <2.50> 0.1 mol/L disodium
dihydrogen ethylenediamine tetraacetate VS until the blue-
purple color of the solution disappears (indicator: 0.5 mg of
phthalein purple): the amount of sulfuric acid (SO4) is 16.0
to 18.0z.
C23H30O6: 402.48
Each mL of 0.1 mol/L barium chloride VS 17,21-Dihydroxypregn-4-ene-3,11,20-trione 21-acetate
＝ 9.606 mg of SO4 [50-04-4]
(2) Related substances—Dissolve 50 mg of Colistin Sul-
Cortisone Acetate, when dried, contains not less
fate in 10 mL of water, and use this solution as the sample
than 97.0z and not more than 102.0z of cortisone
solution. Pipet 1 mL of the sample solution, add water to
acetate (C23H30O6).
solution. Perform the test with these solutions as directed Description Cortisone Acetate occurs as white, crystals or
under Thin-layer Chromatography <2.03>. Spot 1 mL each of crystalline powder.
the sample solution and standard solution on a plate of silica It is sparingly soluble in methanol, slightly soluble in
gel for thin-layer chromatography. Develop the plate with a ethanol (99.5), and practically insoluble in water.
mixture of pyridine, 1-butanol, water and acetic acid (100) Melting point: about 2409 C (with decomposition).
(6:5:4:1) to a distance of about 10 cm, and dry the plate at It shows crystal polymorphism.
1009C for 30 minutes. Spray evenly ninhydrin-butanol TS on
Identification (1) To 2 mg of Cortisone Acetate add 2 mL
the plate, and heat at 1009 C for about 20 minutes: the spot
of sulfuric acid, and allow to stand for a while: a yellowish
other than the principal spot from the sample solution is not
green color is produced, and it gradually changes to yellow-
orange. Examine the solution under ultraviolet light: the so-
Loss on drying <2.41> Not more than 6.0z (1 g, in vacu- lution shows a light green fluorescence. Add carefully 10 mL
um, 609C, 3 hours). of water to this solution: the color of the solution is dis-
charged, and the solution remains clear.
Assay Perform the test according to the Cylinder-plate Cortisone Acetate in methanol (1 in 50,000) as directed un-
method as directed under Microbial Assay for Antibiotics der Ultraviolet-visible Spectrophotometry <2.24>, and com-
<4.02> according to the following conditions. pare the spectrum with the Reference Spectrum or the spec-
(i) Test organism—Escherichia coli NIHJ trum of a solution of Cortisone Acetate RS prepared in the
(ii) Culture medium—Dissolve 10.0 g of peptone, 30.0 g same manner as the sample solution: both spectra exhibit
of sodium chloride, 3.0 g of meat extract and 15.0 g of agar similar intensities of absorption at the same wavelengths.
in 1000 mL of water, adjust the pH with sodium hydroxide (3) Determine the infrared absorption spectrum of Corti-
TS so that the solution will be 6.5 to 6.6 after sterilization, sone Acetate, previously dried, as directed in the potassium
and use as the agar media for seed layer and for base layer. bromide disk method under Infrared Spectrophotometry
(iii) Standard solutions—Weigh accurately an amount of <2.25>, and compare the spectrum with the Reference Spec-
Colistin Sulfate RS, previously dried, equivalent to about trum or the spectrum of previously dried Cortisone Acetate
1,000,000 units, dissolve in phosphate buffer solution (pH RS: both spectra exhibit similar intensities of absorption at
6.0) to make exactly 10 mL, and use this solution as the the same wave numbers. If any difference appears between
standard stock solution. Keep the standard stock solution at the spectra, dissolve Cortisone Acetate and Cortisone Ace-
not exceeding 109C, and use within 7 days. Take exactly a tate RS in acetone, respectively, then evaporate the acetone
suitable amount of the standard stock solution before use, to dryness, and repeat the test on the residues.
add phosphate buffer solution (pH 6.0) to make solutions so
that each mL contains 10,000 units and 2500 units, and use
these solutions as the high concentration standard solution
and the low concentration standard solution, respectively. Purity Related substances—Dissolve 25 mg of Cortisone
(iv) Sample solutions—Weigh accurately an amount of Acetate in 10 mL of a mixture of acetonitrile, water and ace-
Colistin Sulfate, previously dried, equivalent to about tic acid (100) (70:30:1), and use this solution as the sample
1,000,000 units, and dissolve in phosphate buffer solution solution. Pipet 1 mL of the sample solution add the mixture
(pH 6.0) to make exactly 10 mL. Take exactly a suitable of acetonitrile, water and acetic acid (100) (70:30:1) to make
amount of this solution, add phosphate buffer solution (pH exactly 100 mL, and use this solution as the standard solu-
6.0) to make solutions so that each mL contains 10,000 units tion. Perform the test with exactly 15 mL each of the sample
and 2500 units, and use these solutions as the high concen- solution and standard solution as directed under Liquid
tration sample solution and the low concentration sample so- Chromatography <2.01> according to the following condi-
lution, respectively. tions, and determine each peak area by the automatic in-
tegration method: each peak area other than cortisone ace-
tate obtained with the sample solution is not larger than 1/2
times the peak area of cortisone acetate obtained with the
JP XVII Official Monographs / Absorptive Cream 755
cortisone acetate is not larger than 1.5 times the peak area of Column: A stainless steel column 4.6 mm in inside diame-
cortisone acetate with the standard solution. ter and 30 cm in length, packed with octadecylsilanized silica
Operating conditions— gel for liquid chromatography (10 mm in particle diameter).
Detector: An ultraviolet absorption photometer (wave- Column temperature: A constant temperature of about
length: 254 nm). 259C.
Column: A stainless steel column 4.6 mm in inside diame- Mobile phase: A mixture of water and acetonitrile (13:7).
ter and 15 cm in length, packed with octadecylsilanized silica Flow rate: Adjust so that the retention time of cortisone
gel for liquid chromatography (5 mm in particle diameter). acetate is about 12 minutes.
Column temperature: A constant temperature of about System suitability—
259 C. System performance: When the procedure is run with 10
Mobile phase A: A mixture of water and acetonitrile (7:3). mL of the standard solution under the above operating con-
Mobile phase B: A mixture of acetonitrile and water (7:3). ditions, cortisone acetate and the internal standard are eluted
Flowing of mobile phase: Control the gradient by mixing in this order with the resolution between these peaks being
the mobile phases A and B as directed in the following table. not less than 4.
Time after injection Mobile phase A Mobile phase B with 10 mL of the standard solution under the above operat-
of sample (min) (volz) (volz) ing conditions, the relative standard deviation of the ratios
of the peak area of cortisone acetate to that of the internal
0–5 90 10 standard is not more than 1.0z.
5 – 25 90 → 10 10 → 90
25 – 30 10 90
Flow rate: About 1 mL per minute.

Absorptive Cream
retention time of cortisone acetate, beginning after the sol- 吸水クリーム
vent peak.
System suitability— Method of preparation
standard solution add a mixture of acetonitrile, water and White Petrolatum 400 g
acetic acid (100) (70:30:1) to make exactly 10 mL. Confirm Cetanol 100 g
that the peak area of cortisone acetate obtained with 15 mL White Beeswax 50 g
of this solution is equivalent to 8 to 12z of that obtained Sorbitan Sesquioleate 50 g
with 15 mL of the standard solution. Lauromacrogol 5g
System performance: When the procedure is run with 15 Ethyl Parahydroxybenzoate or Methyl
mL of the sample solution under the above operating condi- Parahydroxybenzoate 1g
tions, the number of theoretical plates and the symmetry fac- Butyl Parahydroxybenzoate or Propyl
tor of the peak of cortisone acetate are not less than 10,000 Parahydroxybenzoate 1g
and not more than 1.3, respectively. Purified Water or Purified
System repeatability: When the test is repeated 3 times Water in Containers a sufficient quantity
with 15 mL of the standard solution under the above operat- To make 1000 g
area of cortisone acetate is not more than 5.0z. Melt White Petrolatum, Cetanol, White Beeswax, Sorbi-
tan Sesquioleate and Lauromacrogol by heating on a water
Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, bath, mix and maintain at about 759C. Add Methyl Para-
3 hours). hydroxybenzoate or Ethyl Parahydroxybenzoate and Propyl
Parahydroxybenzoate or Butyl Parahydroxybenzoate to
Purified Water or Purified Water in Containers, dissolve by
Assay Dissolve about 10 mg each of Cortisone Acetate and warming at 809C. Combine both solutions, mix to make
Cortisone Acetate RS, previously dried and accurately emulsion, cool, and stir thoroughly until it congeals.
weighed, in 50 mL of methanol, add exactly 5 mL each of
Description Absorptive Cream is white in color and is
the internal standard solution, then add methanol to make
100 mL, and use these solutions as the sample solution and lustrous. It has a slightly characteristic odor.
the standard solution. Perform the test with 10 mL each of Containers and storage Containers—Tight containers.
area of cortisone acetate to that of the internal standard.
Amount (mg) of cortisone acetate (C23H30O6)
＝ M S × QT / QS
MS: Amount (mg) of Cortisone Acetate RS taken
length: 254 nm).
756 Hydrophilic Cream / Official Monographs JP XVII
velop a yellow color, but neither a brown nor a dark tint.
Hydrophilic Cream Distilling range <2.57> 196 – 2069
C, not less than 90 volz.
親水クリーム Containers and storage Containers—Tight containers.
White Petrolatum 250 g Cresol Solution
Stearyl Alcohol 200 g
Propylene Glycol 120 g クレゾール水
Polyoxyethylene hydrogenated
castor oil 60 40 g
Cresol Solution contains not less than 1.25 volz
Glycerin Monostearate 10 g
and not more than 1.60 volz of cresol.
Methyl Parahydroxybenzoate 1g
Propyl Parahydroxybenzoate 1g Method of preparation
Purified Water or Purified
Saponated Cresol Solution 30 mL
Water, Purified Water or Purified
To make 1000 g Water in Containers a sufficient quantity
Melt White Petrolatum, Stearyl Alcohol, polyoxyethylene To make 1000 mL
hydrogenated castor oil 60 and Glycerin Monostearate by
Prepare by mixing the above ingredients.
heating on a water bath, stir, and keep temperature of the
mixture at about 759 C. To Propylene Glycol add Methyl Description Cresol Solution is a clear or slightly turbid,
Parahydroxybenzoate and Propyl Parahydroxybenzoate, yellow solution. It has the odor of cresol.
melt by warming if necessary, dissolve in Purified Water or
Identification Shake 0.5 mL of the oily layer obtained in
Purified Water in Containers, and warm to about 759C.
the Assay with 30 mL of water, filter, and perform the fol-
Add this solution to the above mixture, stir to form emul-
lowing tests using this filtrate as the sample solution:
sion, cool, and stir thoroughly until it congeals.
(1) To 5 mL of the sample solution add 1 to 2 drops of
Description Hydrophilic Cream is white in color. It has a iron (III) chloride TS: a blue-purple color develops.
slight, characteristic odor. (2) To 5 mL of the sample solution add 1 to 2 drops of
bromine TS: a light yellow, flocculent precipitate is pro-
duced.
Assay Transfer 200 mL of Cresol Solution, exactly meas-
Cresol ured, to a 500-mL distilling flask. Add 40 g of sodium chlo-
ride and 3 mL of dilute sulfuric acid, and connect the distil-
クレゾール ling apparatus with the distilling flask, and distil into a cassia
flask which contains 30 g of powdered sodium chloride and 3
mL of kerosene, exactly measured, until the distillate meas-
C7H8O: 108.14
ures 90 mL. Draw off the water from the condenser, and
continue the distillation until water vapor begins to come out
Cresol is a mixture of isomeric cresols.
of the tip of the condenser. Shake often the cassia flask in
Description Cresol is a clear, colorless or yellow to yellow- warm water to dissolve the sodium chloride, and allow to
brown liquid. It has a phenol-like odor. stand for 15 minutes. After cooling to 159C, add a saturated
It is miscible with ethanol (95) and with diethyl ether. solution of sodium chloride, and allow to stand for more
It is sparingly soluble in water. than 3 hours with occasional shaking. Allow to stand for 1
It dissolves in sodium hydroxide TS. to 2 minutes with gentle shaking to combine the separated oil
A saturated solution of Cresol is neutral to bromocresol drops with the oil layer. The difference between the number
purple TS. of mL of the oil layer measured and 3 mL represents the
It is a highly refractive liquid. amount (mL) of cresol.
It becomes dark brown by light or on aging.
Identification To 5 mL of a saturated solution of Cresol
add 1 to 2 drops of dilute iron (III) chloride TS: a blue-
purple color develops.
20: 1.032 – 1.041
Purity (1) Hydrocarbons—Dissolve 1.0 mL of Cresol in

60 mL of water: the solution shows no more turbidity than
that produced in the following control solution.
Control solution: To 54 mL of water add 6.0 mL of 0.005
mol/L sulfuric acid VS and 1.0 mL of barium chloride TS,
and after thorough shaking, allow to stand for 5 minutes.
(2) Sulfur compounds—Transfer 20 mL of Cresol in a
100-mL conical flask, place a piece of moistened lead (II)
acetate paper on the mouth of the flask, and warm for 5
minutes on a water bath: the lead (II) acetate paper may de-
JP XVII Official Monographs / Croconazole Hydrochloride 757
Allow the cassia flask to stand in warm water for 15 minutes

Saponated Cresol Solution to dissolve the sodium chloride with frequent shaking. Cool
to 159C, add a saturated solution of sodium chloride, and
クレゾール石ケン液 allow to stand for more than 3 hours with occasional shak-
ing. Allow to stand for 1 to 2 minutes with gentle shaking,
and combine the separated oil drops with the oil layer. The
Saponated Cresol Solution contains not less than 42
volume (mL) subtracted 3 (mL) from the oil layer measured
volz and not more than 52 volz of cresol.
represents the amount (mL) of cresol.
Cresol 500 mL Storage—Light-resistant.
Fixed Oil 300 mL
Potassium Hydroxide a suitable quantity
Water, Purified Water or Purified Croconazole Hydrochloride
To make 1000 mL クロコナゾール塩酸塩
Dissolve Potassium Hydroxide, in required quantity for

saponification, in a sufficient quantity of Water, Purified
Water or Purified Water in Containers, add this solution to
fixed oil, previously warmed, add a sufficient quantity of
Ethanol, if necessary, heat in a water bath by thorough
stirring, and continue the saponification. After complete
saponification, add Cresol, stir thoroughly until the mixture
becomes clear, and add sufficient Water, Purified Water or
Purified Water in Containers to make 1000 mL. A corre-
sponding amount of Sodium Hydroxide may be used in C18H15ClN2O.HCl: 347.24
place of Potassium Hydroxide. 1-{1-[2-(3-Chlorobenzyloxy)phenyl]vinyl}-1H-imidazole
monohydrochloride
Description Saponated Cresol Solution is a yellow-brown [77174-66-4]
to red-brown, viscous liquid. It has the odor of cresol.
It is miscible with water, with ethanol (95) and with glyce- Croconazole Hydrochloride, when dried, contains
rin. not less than 98.5z of croconazole hydrochloride
It is alkaline. (C18H15ClN2O.HCl).
Identification Proceed as directed in the Identification un- Description Croconazole Hydrochloride occurs as white to
der Cresol, using the distillate in the Purity (3). pale yellowish white, crystals or crystalline powder.
Purity (1) Alkalinity—Mix well 0.50 mL of Saponated It is very soluble in water, freely soluble in methanol, in
Cresol Solution with 10 mL of neutralized ethanol, add 2 to ethanol (95) and in acetic acid (100), and practically insolu-
3 drops of phenolphthalein TS and 0.10 mL of 1 mol/L hy- ble in diethyl ether.
drochloric acid VS: no red color develops. Identification (1) Determine the absorption spectrum of a
(2) Unsaponified matter—To 1.0 mL of Saponated solution of Croconazole Hydrochloride in methanol (1 in
Cresol Solution add 5 mL of water, and shake: the solution 20,000) as directed under Ultraviolet-visible Spectropho-
is clear. tometry <2.24>, and compare the spectrum with the Refer-
(3) Cresol fraction—Transfer 180 mL of Saponated ence Spectrum: both spectra exhibit similar intensities of ab-
Cresol Solution to a 2000-mL distilling flask, add 300 mL of sorption at the same wavelengths.
water and 100 mL of dilute sulfuric acid, and distil with (2) Determine the infrared absorption spectrum of
steam until the distillate becomes clear. Draw off the water Croconazole Hydrochloride, previously dried, as directed in
from the condenser, and continue the distillation until water the potassium chloride disk method under Infrared Spectro-
vapor begins to come out of the tip of the condenser. Cool photometry <2.25>, and compare the spectrum with the Ref-
the condenser again, and continue distillation for 5 minutes. erence Spectrum: both spectra exhibit similar intensities of
Dissolve 20 g of sodium chloride per 100 mL of the distillate, absorption at the same wave numbers.
allow to stand, and collect the separated clear oil layer. After (3) Dissolve 0.05 g of Croconazole Hydrochloride in 10
adding about 15 g of powdered calcium chloride for drying mL of water, add 2 mL of sodium hydroxide TS and 20 mL
in small portions with frequent shaking, allow to stand for 4 of diethyl ether, and shake. Wash the separated aqueous
hours. Filter, and distil exactly 50 mL of the filtrate: the dis- layer with two 10-mL portions of diethyl ether, and acidify
tillate is not less than 43 mL between 1969 C and 2069C. the solution with 2 mL of dilute nitric acid: the solution re-
Assay Transfer 5 mL of Saponated Cresol Solution, ex- sponds to the Qualitative Tests <1.09> for chloride.
actly measured, to a 500-mL distilling flask, holding the Melting point <2.60> 148 – 1539C
pipet vertically for 15 minutes to draw off the solution into
the flask. Add 200 mL of water, 40 g of sodium chloride and Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
3 mL of dilute sulfuric acid, connect the distilling apparatus Croconazole Hydrochloride according to Method 4, and per-
with the distilling flask, and distil into a cassia flask which form the test. Prepare the control solution with 1.0 mL of
contains 30 g of powdered sodium chloride and exactly 3 mL Standard Lead Solution (not more than 10 ppm).
of kerosene, until the distillate reaches 90 mL. Draw off the (2) Related substances—Dissolve 50 mg of Croconazole
water from the condenser, and continue the distillation until Hydrochloride in 10 mL of methanol, and use this solution
water vapor begins to come out of the tip of the condenser. as the sample solution. Pipet 1 mL of the sample solution,
add methanol to make exactly 100 mL, and use this solution
758 Crospovidone / Official Monographs JP XVII
as the standard solution. Perform the test with these solu- the sieve in a drying machine at 1059C for 5 hours without
tions as directed under Thin-layer Chromatography <2.03>. air-circulation. After cooling down in a desiccator for 30
Spot 10 mL each of the sample solution and standard solu- minutes, weigh the mass of the residue with sieve, and calcu-
tion on a plate of silica gel with fluorescent indicator for late the amount of the residue on the sieve by the following
thin-layer chromatography. Develop the plate with a mixture equation: Type A is more than 15z, and type B is not more
of ethyl acetate, hexane, methanol and ammonia solution than 15z.
(28) (30:15:5:1) to a distance of about 10 cm, and air-dry the
Amount (z) of the residue of Crospovidone on
No. 235 (63 mm) sieve
nm): the spots other than the principal spot and the spot of
＝ (M1 － M3)/M2 × 100
the starting point from the sample solution are not more in-
tense than the spot from the standard solution. M1: The mass (g) of the residue with sieve after 5 hours
drying
M2: Amount (g) of Crospovidone taken, calculated on the
4 hours).
dried basis
Residue on ignition <2.44> Not more than 0.1z (1 g). M3: Mass (g) of the sieve
Assay Weigh accurately about 0.6 g of Croconazole Hy- Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
drochloride, previously dried, dissolve in 10 mL of acetic Crospovidone according to Method 2, and perform the test.
acid (100), add 40 mL of acetic anhydride, and titrate <2.50> Prepare the control solution with 2.0 mL of Standard Lead
with 0.1 mol/L perchloric acid VS [indicator: 1 to 2 drops of Solution (not more than 10 ppm).
a solution of malachite green oxalate in acetic acid (100) (1 in (2) Water-soluble substances—Place 25.0 g of Crospovi-
100)] until the color of the solution changes from blue-green done in a 400-mL beaker, add 200 mL of water, and stir for
through green to yellow-green. Perform a blank determina- 1 hour. Transfer the suspension to a 250-mL volumetric
tion, and make any necessary correction. flask, rinsing with water, and dilute to volume with water.
Allow the bulk of the solids to settle. Filter about 100 mL of
the almost clear supernatant liquid through a 0.45 mm mem-
＝ 34.72 mg of C18H15ClN2O.HCl
brane filter, protected by superimposing a 3 mm membrane
Containers and storage Containers—Tight containers. filter. Transfer exactly 50 mL of the clear filtrate to a tared
Storage—Light-resistant. 100-mL beaker, evaporate to dryness and dry at 105 – 1109 C
for 3 hours: the mass of the residue is not more than 75 mg.
(3) 1-Vinyl-2-pyrrolidone—To 1.250 g of Crospovidone
Crospovidone add exactly 50 mL of methanol, and shake for 60 minutes.
Leave bulk to settle, filter through a 0.2 mm membrane filter,
クロスポビドン and use the filtrate as the sample solution. Separately, dis-
solve 50 mg of 1-vinyl-2-pyrrolidone in methanol to make
exactly 100 mL. Pipet 1 mL of this solution, and add metha-
nol to make exactly 100 mL. To exactly 5 mL of this solution
Crospovidone is a cross-linked polymer of 1-vinyl-2-
pyrrolidone.
according to the following conditions: the peak area of 1-
vinyl-2-pyrrolidone obtained from the sample solution is not
12.8z of nitrogen (N: 14.01), calculated on the dried
larger than that obtained from the standard solution (not
basis.
more than 10 ppm).
Two types of Crospovidone are available, depend-
ing on the particle size: type A and type B.
The label states the type. Detector: An ultraviolet absorption photometer (wave-

length: 235 nm).

Description Crospovidone occurs as a white to pale yel- Column: Two stainless steel columns, one is 4 mm in in-
lowish powder. side diameter and 25 mm in length and the other is 4 mm in
It is practically insoluble in water, in methanol and in inside diameter and 250 mm in length, they are packed with
ethanol (99.5). octadecylsilanized silica gel for liquid chromatography (5 mm
It is hygroscopic. in particle diameter), and used them as the pre-column and
the separation column, respectively.
Identification (1) Suspend 1 g of Crospovidone in 10 mL
of water, add 0.1 mL of iodine TS, shake for 30 seconds,
409C.
then add 1 mL of starch TS, and shake: a blue color is not
Mobile phase: A mixture of water and acetonitrile (9:1).
produced within 30 seconds.
(2) When add 0.1 g of Crospovidone to 10 mL of water,
Washing of pre-column: After each injection of the sam-
shake to suspend, and allow the suspension to stand, a clear
ple solution, wash the pre-column by passing the mobile
liquid is not produced within 15 minutes.
phase backwards, at the same flow rate as applied in the test,
Particle size Weigh accurately about 20 g of Crospovidone, for 30 minutes.
place in a 1000-mL conical flask, add 500 mL of water, System suitability—
shake for 30 minutes, and pour onto an accurately tared No. System performance: Dissolve 10 mg of 1-vinyl-2-pyrroli-
235 (63 mm) sieve, previously washed with hot water and done and 0.50 g of vinyl acetate in methanol to make 100
dried at 1059C for a night, and wash the residue with water mL. To 1 mL of this solution add the mobile phase to make
until the passing water is clear. Dry the residue together with 100 mL. When the procedure is run with 50 mL of this solu-
JP XVII Official Monographs / Cyanamide 759
tion under the above operating conditions, 1-vinyl-2-pyrroli-

done and vinyl acetate are eluted in this order with the reso- Cyanamide
lution between these peaks being not less than 2.0.
System repeatability: When the test is repeated 6 times シアナミド
area of 1-vinyl-2-pyrrolidone is not more than 2.0z.
CH2N2: 42.04
(4) Peroxides—
Aminonitrile
Method 1: Apply to the sample labeled as type A. Suspend
[420-04-2]
4.0 g of Crospovidone in 100 mL of water, and use as the
sample suspension. To 25 mL of the sample suspension add
Cyanamide contains not less than 97.0z and not
2 mL of titanium (III) chloride-sulfuric acid TS, allow to
more than 101.0z of cyanamide (CH2N2), calculated
stand for 30 minutes, and filter. Determine the absorbance
on the anhydrous basis.
of the filtrate at 405 nm as directed under Ultraviolet-visible
Spectrophotometry <2.24>, using the control, prepared by Description Cyanamide occurs as white, crystals or crystal-
filtrating the sample suspension and adding 2 mL of diluted line powder.
sulfuric acid (13 in 100) to 25 mL of this filtrate: not more It is very soluble in water, in methanol, in ethanol (99.5)
than 0.35 (not more than 400 ppm expressed as hydrogen and in acetone.
peroxide). The pH of a solution of 1.0 g of Cyanamide in 100 mL of
Method 2: Apply to the sample labeled as type B. Suspend water is between 5.0 and 6.5.
2.0 g of Crospovidone in 50 mL of water, and use as the It is hygroscopic.
sample suspension. To 10 mL of the sample suspension add Melting point: about 469C
water to make 25 mL, add 2 mL of titanium (III) chloride-
Identification (1) To 1 mL of a solution of Cyanamide
sulfuric acid TS, allow to stand for 30 minutes, and filter.
(1 in 100) add 1 mL of potassium 1,2-naphthoquinone-4-
Determine the absorbance of the filtrate at 405 nm as di-
sulfonate TS and 0.2 mL of sodium hydroxide TS: a deep
red color develops.
using the control, prepared by filtrating the sample suspen-
(2) Drop one or two drops of a solution of Cyanamide in
sion, adding water to 10 mL of this filtrate to make 25 mL
acetone (1 in 100) onto a potassium bromide disk prepared
and 2 mL of diluted sulfuric acid (13 in 100): not more than
as directed in the potassium bromide disk method under In-
0.35 (not more than 1000 ppm expressed as hydrogen
frared Spectrophotometry <2.25>, and air-dry the disk. De-
peroxide).
termine the infrared absorption spectrum of the disk as di-
Loss on drying <2.41> Not more than 5.0z (0.5 g, 1059C, rected in the film method under Infrared Spectrophotometry
constant mass). <2.25>, and compare the spectrum with the Reference Spec-
Assay Weigh accurately about 0.1 g of Crospovidone,
place in a Kjeldahl flask, add 5 g of a powdered mixture of
of Cyanamide in 10 mL of water: the solution is clear and
33 g of potassium sulfate, 1 g of copper (II) sulfate pentahy-
colorless.
drate and 1 g of titanium (IV) oxide, and 3 glass beads.
Wash any adhering particles from the neck into the flask
Cyanamide according to Method 1, and perform the test.
with a small quantity of water. Add 7 mL of sulfuric acid,
allowing it to run down the inside wall of the flask. Gradual-
ly heat the flask until the solution has a clear, yellowish-
green color, and the inside wall of the flask is free from Water <2.48> Not more than 1.0z (1 g, volumetric titra-
carbonized material, and then heat for a further 45 minutes. tion, direct titration).
After cooling, cautiously add 20 mL of water, and connect
the flask to the distillation apparatus previously washed by
passing steam through it. To the absorption flask add 30 mL Assay Weigh accurately about 1 g of Cyanamide, and dis-
of a solution of boric acid (1 in 25), 3 drops of bromocresol solve in water to make exactly 250 mL. Pipet 15 mL of this
green-methyl red TS and sufficient water to immerse the solution, add 2 to 3 drops of dilute nitric acid, 10 mL of am-
lower end of the condenser tube. Add 30 mL of a solution of monia TS and exactly 50 mL of 0.1 mol/L silver nitrate VS,
sodium hydroxide (21 in 50) through a funnel, cautiously and allow to stand for 15 minutes with occasional shaking.
rinse the funnel with 10 mL of water, immediately close the Add water to make exactly 100 mL, filter, discard the first
clamp attached to the rubber tube, then start the distillation 20 mL of the filtrate, and pipet 50 mL of the subsequent
with steam to obtain 80 – 100 mL of distillate. Remove the filtrate. After neutralizing this solution with dilute nitric
absorption flask from the lower end of the condenser tube, acid, add 3 mL of dilute nitric acid, and titrate <2.50> the
rinsing the end part with a small quantity of water, and excess silver nitrate with 0.1 mol/L ammonium thiocyanate
titrate <2.50> the distillate with 0.025 mol/L sulfuric acid VS VS (indicator: 2 mL of ammonium iron (III) sulfate TS).
until the color of the solution changes from green through Perform a blank determination.
pale grayish-blue to pale grayish red-purple. Carry out a
blank determination and make any necessary correction.
＝ 2.102 mg of CH2N2
＝ 0.7003 mg of N
760 Cyanocobalamin / Official Monographs JP XVII
add dropwise diluted sulfuric acid (1 in 7) until a clear solu-
Cyanocobalamin tion results, then add 3 to 5 drops more of diluted sulfuric
acid (1 in 7): a blue to blue-green color develops.
Vitamin B12 pH <2.54> Dissolve 0.10 g of Cyanocobalamin in 20 mL of
シアノコバラミン
Purity (1) Clarity and color of solution—Dissolve 20 mg
of Cyanocobalamin in 10 mL of water: the solution is clear
and red in color.
light-resistant vessels. Dissolve 10 mg of Cyanocobalamin in
10 mL of the mobile phase, and use this solution as the sam-
ple solution. Pipet 3 mL of the sample solution, add the mo-
tomatic integration method: the total area of the peak other
than cyanocobalamin obtained from the sample solution is
not larger than the peak area of cyanocobalamin obtained
length: 361 nm).
C63H88CoN14O14P: 1355.37
Coa-[a-(5,6-Dimethyl-1H-benzimidazol-1-yl)]-Cob-
cyanocobamide
[68-19-9]
309C.
Mobile phase: Dissolve 10 g of anhydrous disodium
Cyanocobalamin contains not less than 96.0z
hydrogen phosphate in 1000 mL of water, and adjust to pH
and not more than 102.0z of cyanocobalamin
3.5 with phosphoric acid. To 147 mL of this solution add 53
(C63H88CoN14O14P), calculated on the dried basis.
mL of methanol.
Description Cyanocobalamin occurs as dark red, crystals Flow rate: Adjust so that the retention time of
or powder. cyanocobalamin is about 7 minutes.
It is sparingly soluble in water, and slightly soluble in Time span of measurement: About 4 times as long as the
ethanol (99.5). retention time of cyanocobalamin, beginning after the sol-
It is hygroscopic. vent peak.
Identification (1) Determine the absorption spectrum of
lution, add the mobile phase to make 100 mL, and use this
of a solution of Cyanocobalamin RS prepared in the same
area of cyanocobalamin obtained with 20 mL of this solution
is equivalent to 7 to 13z of that obtained with 20 mL of the
(2) Mix 1 mg of Cyanocobalamin with 50 mg of potas-
sium hydrogen sulfate, and fuse by igniting. Cool, break up
System performance: Perform this procedure quickly after
the mass with a glass rod, add 3 mL of water, and dissolve
the solution is prepared. To 25 mg of cyanocobalamin add
by boiling. Add 1 drop of phenolphthalein TS, then add
10 mL of water, and warm, if necessary, to dissolve. After
dropwise sodium hydroxide TS until a light red color just
cooling, add 0.5 mL of sodium toluenesulfonchloramide TS,
develops. Add 0.5 g of sodium acetate trihydrate, 0.5 mL of
0.5 mL of 0.05 mol/L hydrochloric acid TS and water to
dilute acetic acid and 0.5 mL of a solution of disodium 1-
make 25 mL, mix, and allow the solution to stand for 5
nitroso-2-naphthol-3,6-disulfonate (1 in 500): a red to
minutes. To 1 mL of the solution add the mobile phase to
orange-red color is immediately produced. Then add 0.5 mL
make 10 mL. When the procedure is run with 20 mL of the
of hydrochloric acid, and boil for 1 minute: the red color
solution under the above operating conditions, two principal
does not disappear.
peaks appear with the resolution between these peaks being
(3) Transfer 5 mg of Cyanocobalamin to a 50-mL distil-
not less than 2.5.
ling flask, dissolve in 5 mL of water, and add 2.5 mL of
hypophosphorous acid. Connect the flask with a short con-
denser, and dips its tip into a test tube containing 1 mL of a
solution of sodium hydroxide (1 in 50). Heat gently for 10
tion of the peak area of cyanocobalamin is not more than
minutes, then distil 1 mL into a test tube. To the test tube
3.0z.
add 4 drops of a saturated solution of ammonium iron (II)
sulfate hexahydrate, shake gently, then add about 30 mg of Loss on drying <2.41> Not more than 12z (50 mg, in vacu-
sodium fluoride, and heat the contents to boil. Immediately um at a pressure not exceeding 0.67 kPa, phosphorus (V)
JP XVII Official Monographs / Cyclopentolate Hydrochloride 761
oxide, 1009C, 4 hours). MS: Amount (mg) of Cyanocobalamin RS taken, calcu-

lated on the dried basis
Assay Weigh accurately about 20 mg each of Cyanocobala-
min and Cyanocobalamin RS (previously determine the loss Containers and storage Containers—Hermetic containers,
on drying <2.41> under the same conditions as Cyanocobala- and colored containers may be used.
min), dissolve in water to make exactly 1000 mL, respec- Storage—Light-resistant.
tively, and use these solutions as the sample solution and the
of the sample solution and standard solution, at 361 nm as Cyclopentolate Hydrochloride
directed under Ultraviolet-visible Spectrophotometry <2.24>.
シクロペントラート塩酸塩
Amount (mg) of cyanocobalamin (C63H88CoN14O14P)
＝ M S × AT / AS
MS: Amount (mg) of Cyanocobalamin RS taken, calcu-
lated on the dried basis
C17H25NO3.HCl: 327.85
2-(Dimethylamino)ethyl (2RS )-2-(1-
Cyanocobalamin Injection hydroxycyclopentyl)phenylacetate monohydrochloride
[5870-29-1]
Vitamin B12 Injection
Cyclopentolate Hydrochloride, when dried, con-
シアノコバラミン注射液 tains not less than 98.5z of cyclopentolate hydrochlo-
ride (C17H25NO3.HCl).
Cyanocobalamin Injection is an aqueous injection. Description Cyclopentolate Hydrochloride occurs as a
It contains not less than 95.0z and not more than white crystalline powder. It is odorless, or has a characteris-
115.0z of the labeled amount of cyanocobalamin tic odor.
(C63H88CoN14O14P: 1355.37). It is very soluble in water, freely soluble in ethanol (95), in
acetic acid (100) and in chloroform, sparingly soluble in ace-
tic anhydride, and practically insoluble in diethyl ether.
tions, with Cyanocobalamin.
Identification (1) To 1 mL of a solution of Cyclopento-
Description Cyanocobalamin Injection is a clear, light red
late Hydrochloride (1 in 100) add 1 mL of Reinecke salt TS:
to red liquid.
a light red precipitate is formed.
Identification Determine the absorption spectrum of the (2) Dissolve 0.2 g of Cyclopentolate Hydrochloride in 2
sample solution obtained in the Assay as directed under Ul- mL of water, add 2 mL of sodium hydroxide TS, and boil
traviolet-visible Spectrophotometry <2.24>: it exhibits maxi- for 1 minute. After cooling, add 2 drops of nitric acid: a
ma between 277 nm and 279 nm, between 360 nm, and 362 phenylacetic acid-like odor is perceptible.
nm and between 548 nm and 552 nm. Determine the absor- (3) Determine the infrared absorption spectrum of Cy-
bances, A1 and A2, of this solution at the wavelengths of clopentolate Hydrochloride, previously dried, as directed in
maximum absorption between 360 nm and 362 nm, and be- the potassium chloride disk method under Infrared Spectro-
tween 548 nm and 552 nm, respectively: the ratio A2/A1 is photometry <2.25>, and compare the spectrum with the Ref-
not less than 0.29 and not more than 0.32. erence Spectrum: both spectra exhibit similar intensities of
(4) A solution of Cyclopentolate Hydrochloride (1 in 50)
Extractable volume <6.05> It meets the requirement. responds to the Qualitative Tests <1.09> for chloride.
Foreign insoluble matter <6.06> Perform the test according pH <2.54> Dissolve 0.20 g of Cyclopentolate Hydrochlo-
to Method 1: it meets the requirement. ride in 20 mL of water: the pH of this solution is between 4.5
and 5.5.
ment. Melting point <2.60> 135 – 1389C
Sterility <4.06> Perform the test according to the Mem- Purity (1) Clarity and color of solution—Dissolve 1.0 g
brane filtration method: it meets the requirement. of Cyclopentolate Hydrochloride in 10 mL of water: the so-
lution is clear and colorless.
Assay Measure exactly a volume of Cyanocobalamin
(2) Heavy metals <1.07>—Proceed with 1.0 g of Cy-
Injection, equivalent to about 2 mg of cyanocobalamin
clopentolate Hydrochloride according to Method 1, and per-
(C63H88CoN14O14P), add water to make exactly 100 mL, and
accurately about 20 mg of Cyanocobalamin RS (previously
(3) Related substances—Dissolve 0.20 g of Cyclopento-
determine the loss on drying <2.41> under the same condi-
late Hydrochloride in 10 mL of chloroform, and use this so-
tions as Cyanocobalamin), add water to make exactly 1000
mL, and use this solution as the standard solution. Then,
tion, and add chloroform to make exactly 20 mL. Pipet 1
proceed as directed in the Assay under Cyanocobalamin.
mL of this solution, add chloroform to make exactly 10 mL,
Amount (mg) of cyanocobalamin (C63H88CoN14O14P) and use this solution as the standard solution. Perform the
＝ MS × AT/AS × 1/10 test with these solutions as directed under Thin-layer Chro-
762 Cyclophosphamide Hydrate / Official Monographs JP XVII
matography <2.03>. Spot 10 mL each of the sample solution of Cyclophosphamide Hydrate in 10 mL of water: the solu-
and standard solution on a plate of silica gel for thin-layer tion is clear and colorless.
chromatography. Develop the plate with a mixture of 2- (2) Chloride <1.03>—Perform the test with 0.40 g of Cy-
propanol, n-butyl acetate, water and ammonia solution (28) clophosphamide Hydrate at a temperature not exceeding
(100:60:23:17) to a distance of about 10 cm, and air-dry the 209C. Prepare the control solution with 0.40 mL of 0.01
plate. Spray evenly a solution of sulfuric acid in ethanol mol/L hydrochloric acid VS (not more than 0.036z).
(99.5) (1 in 10) on the plate, and heat at 1209C for 30 (3) Heavy metals <1.07>—Proceed with 1.0 g of Cy-
minutes. Examine under ultraviolet light (main wavelength: clophosphamide Hydrate according to Method 1, and per-
254 nm): the spots other than the principal spot from the form the test. Prepare the control solution with 2.0 mL of
sample solution are not more intense than the spot from the Standard Lead Solution (not more than 20 ppm).
standard solution.
Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, titration).
4 hours).
Assay Weigh accurately about 0.3 g of Cyclophosphamide
Residue on ignition <2.44> Not more than 0.05z (1 g). Hydrate, add 15 mL of hydrogen chloride-ethanol TS, and
heat in a water bath under a reflux condenser for 3.5 hours
Assay Weigh accurately about 0.5 g of Cyclopentolate Hy-
while protecting from moisture. Distil the ethanol under
reduced pressure. Dissolve the residue in 40 mL of a mixture
<2.50> with 0.1 mol/L perchloric acid-dioxane VS (indicator:
the solution changes from purple through blue-green to yel-
2 drops of crystal violet TS) until the color of the solution
low-green (indicator: 2 drops of crystal violet TS). Perform a
changes from blue through green to yellow. Perform a blank
＝ 13.96 mg of C7H15Cl2N2O2P.H2O
Storage—Not exceeding 309C.
Cyclophosphamide Hydrate
シクロホスファミド水和物 Cyclophosphamide Tablets
シクロホスファミド錠
Cyclophosphamide Tablets contain not less than

93.0z and not more than 107.0z of the labeled amount
of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O:
C7H15Cl2N2O2P.H2O: 279.10
279.10).
N, N-Bis(2-chloroethyl)-3,4,5,6-tetrahydro-2H-1,3,2-
oxazaphosphorin-2-amine 2-oxide monohydrate Method of preparation Prepare as directed under Tablets,
[6055-19-2] with Cyclophosphamide Hydrate.
Identification To Cyclophosphamide Tablets add 1 mL of
Cyclophosphamide Hydrate contains not less than
water for every 53 mg of Cyclophosphamide Hydrate, shake
97.0z of cyclophosphamide hydrate (C7H15Cl2N2O2P.
vigorously for 5 minutes, add 6 mL of methanol for every 53
H2O).
mg of Cyclophosphamide Hydrate, and shake vigorously for
Description Cyclophosphamide Hydrate occurs as white, 10 minutes. To this solution add methanol so that each mL
crystals or crystalline powder. It is odorless. contains about 5.3 mg of Cyclophosphamide Hydrate, and
It is very soluble in acetic acid (100), freely soluble in centrifuge. Filter the supernatant liquid through a membrane
ethanol (95), in acetic anhydride and in chloroform, and filter with a pore size not exceeding 0.45 mm. Discard not less
soluble in water and in diethyl ether. than 3 mL of the first filtrate, and use the subsequent filtrate
Melting point: 45 – 539C as the sample solution. Separately, dissolve 53 mg of cy-
clophosphamide hydrate for assay in 10 mL of a mixture of
Identification (1) Dissolve 0.1 g of Cyclophosphamide
methanol and water (9:1), and use this solution as the stand-
Hydrate in 10 mL of water, and add 5 mL of silver nitrate
ard solution. Perform the test with these solutions as di-
TS: no precipitate is produced. Then boil this solution: a
rected under Thin-layer Chromatography <2.03>. Spot 2 mL
white precipitate is produced. Collect the precipitate, and
each of the sample solution and standard solution on a plate
add dilute nitric acid to a portion of this precipitate: it does
of silica gel for thin-layer chromatography. Develop the
not dissolve. Add excess ammonia TS to another portion of
plate with a mixture of 1-propanol and water (8:1) to a dis-
the precipitate: it dissolves.
tance of about 10 cm, and air-dry the plate. Heat the plate at
(2) Add 1 mL of diluted sulfuric acid (1 in 25) to 0.02 g
1309C for 15 minutes. After cooling, spray evenly ninhydrin-
of Cyclophosphamide Hydrate, and heat until white fumes
butanol TS on the plate, and after air-drying heat at 1309C
are evolved. After cooling, add 5 mL of water, and shake.
for 10 minutes: the principal spot obtained from the sample
Neutralize with ammonia TS, then acidify with dilute nitric
solution and the spot obtained from the standard solution
acid: this solution responds to the Qualitative Tests <1.09>
show a red-purple color and the same Rf value.
(2) for phosphate.
JP XVII Official Monographs / Cycloserine 763
Content uniformity test. of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O), and

To 1 tablet of Cyclophosphamide Tablets add 3V/5 mL of centrifuge. Filter the supernatant liquid through a membrane
a mixture of water and methanol (3:2), and shake vigorously filter with a pore size not exceeding 0.45 mm. Discard the
to homogenously disperse the tablet. To this solution add a first 3 mL of the filtrate, pipet 4 mL of the subsequent fil-
mixture of water and methanol (3:2) to make exactly V mL trate, add a mixture of water and methanol (3:2) to make ex-
so that each mL contains about 1.1 mg of cyclophosphamide actly 10 mL, and use this solution as the sample solution.
hydrate (C7H15Cl2N2O2P.H2O), and centrifuge. Filter the Separately, weigh accurately about 53 mg of cyclophos-
supernatant liquid through a membrane filter with a pore phamide hydrate for assay, dissolve in a mixture of water
size not exceeding 0.45 mm. Discard the first 3 mL of the fil- and methanol (3:2) to make exactly 50 mL, and use this solu-
trate, and use the subsequent filtrate as the sample solution. tion as the standard solution. Perform the test with exactly
Then, proceed as directed in the Assay. 20 mL each of the sample solution and standard solution as
Amount (mg) of cyclophosphamide hydrate
the following conditions, and determine the peak area, AT
(C7H15Cl2N2O2P.H2O)
and AS, of cyclophosphamide in each solution.
＝ MS × AT/AS × V/50
Amount (mg) of cyclophosphamide hydrate
MS: Amount (mg) of cyclophosphamide hydrate for assay
(C7H15Cl2N2O2P.H2O)
taken
＝ MS × AT/AS × V/200
lutions per minute according to the Basket method, using
taken
900 mL of water as the dissolution medium, the dissolution
rate in 45 minutes of Cyclophosphamide Tablets is not less Operating conditions—
than 80z. Detector: An ultraviolet absorption photometer (wave-
Start the test with 1 tablet of Cyclophosphamide Tablets, length: 205 nm).
withdraw not less than 20 mL of the medium at the specified Column: A stainless steel column 4.6 mm in inside diame-
minute after starting the test, and filter through a membrane ter and 15 cm in length, packed with octadecylsilanized silica
filter with a pore size not exceeding 0.45 mm. Discard the gel for liquid chromatography (5 mm in particle diameter).
first 10 mL of the filtrate, pipet V mL of the subsequent Column temperature: A constant temperature of about
filtrate, add water to make exactly V? mL so that each mL 259C.
contains about 59 mg of cyclophosphamide hydrate Mobile phase: A mixture of water and methanol (3:2).
(C7H15Cl2N2O2P.H2O) and use this solution as the sample so- Flow rate: Adjust so that the retention time of cy-
lution. Separately, weigh accurately about 30 mg of cyclophosphamide is about 10 minutes.
clophosphamide hydrate for assay, and dissolve in water to System suitability—
make exactly 50 mL. Pipet 2 mL of this solution, add water System performance: When the procedure is run with 20
to make exactly 20 mL, and use this solution as the standard mL of the standard solution under the above operating con-
solution. Perform the test with exactly 50 mL each of the ditions, the number of theoretical plates and the symmetry
sample solution and standard solution as directed under factor of the peak of cyclophosphamide are not less than
Liquid Chromatography <2.01> according to the following 4000 and not more than 1.5, respectively.
conditions and determine the peak areas, AT and AS, of System repeatability: When the test is repeated 6 times
cyclophosphamide in each solution. with 20 mL of the standard solution under the above operat-
area of cyclophosphamide is not more than 1.0z.
of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O)
＝ MS × AT/AS × V?/V × 1/C × 180 Containers and storage Containers—Tight containers.
taken
C: Labeled amount (mg) of cyclophosphamide hydrate Cycloserine
(C7H15Cl2N2O2P.H2O) in 1 tablet
サイクロセリン
Assay.
mL of the standard solution under the above operating con- C3H6N2O2: 102.09
ditions, the number of theoretical plates and the symmetry (4R)-4-Aminoisoxazolidin-3-one
factor of the peak of clophosphamide are not less than 5000 [68-41-7]
System repeatability: When the test is repeated 6 times Cycloserine contains not less than 950 mg (potency)
with 50 mL of the standard solution under the above operat- and not more than 1020 mg (potency) per mg, calcu-
ing conditions, the relative standard deviation of the peak lated on the dried basis. The potency of Cycloserine is
area of clophosphamide is not more than 2.0z. expressed as mass (potency) of cycloserine (C3H6N2O2).
Assay To 10 tablets of Cyclophosphamide Tablets add Description Cycloserine occurs as white to light yellowish
13V/20 mL of a mixture of water and methanol (3:2), and white, crystals or crystalline powder.
shake vigorously to homogenously disperse the tablets. To It is soluble in water, and sparingly soluble in ethanol (95).
this solution add a mixture of water and methanol (3:2) to
of Cycloserine, previously dried, as directed in the potassium
764 Cyproheptadine Hydrochloride Hydrate / Official Monographs JP XVII
<2.25>, and compare the spectrum with the Reference Spec- Cyproheptadine Hydrochloride
trum or the spectrum of previously dried Cycloserine RS:
both spectra exhibit similar intensities of absorption at the Hydrate
same wave numbers.
シプロヘプタジン塩酸塩水和物
Optical rotation <2.49> [a]20D : ＋108 – ＋1149(2.5 g calcu-
lated on the dried basis, 2 mol/L sodium hydroxide TS, 50
mL, 100 mm).
pH <2.54> Dissolve 1.0 g of Cycloserine in 20 mL of water:
Cycloserine according to Method 4, and perform the test.
C21H21N.HCl.1 1/2 H2O: 350.88
4-(5H-Dibenzo[a,d ]cyclohepten-5-ylidene)-1-
(2) Condensation products—Dissolve 20 mg of Cycloser-
methylpiperidine monohydrochloride sesquihydrate
ine in sodium hydroxide TS to make exactly 50 mL, and de-
[41354-29-4]
termine the absorbance of this solution at 285 nm as directed
under Ultraviolet-visible Spectrophotometry <2.24>: not
Cyproheptadine Hydrochloride Hydrate, when
more than 0.8.
dried, contains not less than 98.5z of cyproheptadine
Loss on drying <2.41> Not more than 1.5z (0.5 g, reduced hydrochloride (C21H21N.HCl: 323.86).
C, 3 hours).
pressure, 609
Description Cyproheptadine Hydrochloride Hydrate oc-
Residue on ignition <2.44> Not more than 0.5z (1 g). curs as a white to pale yellow crystalline powder. It is odor-
less, and has a slightly bitter taste.
It is freely soluble in methanol and in acetic acid (100),
soluble in chloroform, sparingly soluble in ethanol (95),
slightly soluble in water, and practically insoluble in diethyl
(i) Test organism—Bacillus subtilis ATCC 6633
ether.
(ii) Culture medium—Use the medium i in 1) Medium
for test organism [5] under (1) Agar media for seed and base Identification (1) Dissolve 0.1 g of Cyproheptadine Hy-
layer. Adjust the pH of the medium so that it will be 6.0 to drochloride Hydrate in 10 mL of methanol, apply 1 drop of
6.1 after sterilization. this solution on filter paper, air-dry, and examine under ul-
(iii) Standard solutions—Weigh accurately an amount of traviolet light (main wavelength: 254 nm): the solution shows
Cycloserine RS, previously dried at 609 C for 3 hours under a pale blue fluorescence.
reduced pressure of not exceeding 0.67 kPa, equivalent to (2) Weigh 0.1 g of Cyproheptadine Hydrochloride Hy-
about 40 mg (potency), dissolve in water to make exactly 100 drate, transfer to a separator, dissolve in 5 mL of chlo-
mL, and use this solution as the standard stock solution. roform, add 4 mL of water and 1 mL of sodium carbonate
Keep the standard stock solution at 59 C or below and use TS, and shake. Transfer the chloroform layer to another
within 24 hours. Take exactly a suitable amount of the separator, and wash with 4 mL of water by shaking well.
standard stock solution before use, and add phosphate Filter the chloroform layer through absorbent cotton
buffer solution (pH 6.0) to make solutions so that each mL moistened previously with chloroform, and evaporate the
contains 100 mg (potency) and 50 mg (potency), and use these filtrate to dryness. Dissolve the residue in 8 mL of dilute
solutions as the high concentration standard solution and the ethanol by warming at 659C. Rub the inner wall of the con-
low concentration standard solution, respectively. tainer with a glass rod while cooling until crystallization be-
(iv) Sample solutions—Weigh accurately an amount of gins, and allow to stand for 30 minutes. Collect the crystals,
Cycloserine equivalent to about 40 mg (potency), dissolve in and dry at 809C for 2 hours: the crystals melt <2.60> between
water to make exactly 100 mL. Take exactly a suitable 1119C and 1159C.
amount of this solution, add phosphate buffer solution (pH (3) Determine the absorption spectrum of a solution of
6.0) to make solutions so that each mL contains 100 mg (po- Cyproheptadine Hydrochloride Hydrate in ethanol (95) (1 in
tency) and 50 mg (potency), and use these solutions as the 100,000) as directed under Ultraviolet-visible Spectropho-
high concentration sample solution and the low concentra- tometry <2.24>, and compare the spectrum with the Refer-
tion sample solution, respectively. ence Spectrum: both spectra exhibit similar intensities of ab-
(4) A saturated solution of Cyproheptadine Hydrochlo-
ers.
ride Hydrate responds to the Qualitative Tests <1.09> (2) for
chloride.
Purity (1) Acidity—Dissolve 2.0 g of Cyproheptadine
Hydrochloride Hydrate in 25 mL of methanol, and add 1
drop of methyl red TS and 0.30 mL of 0.1 mol/L sodium hy-
droxide VS: a yellow color develops.
(2) Heavy metals <1.07>—Proceed with 1.0 g of Cypro-
heptadine Hydrochloride Hydrate according to Method 2,
and perform the test. Prepare the control solution with 2.0
Loss on drying <2.41> 7.0 – 9.0z (1 g, in vacuum at a pres-
JP XVII Official Monographs / L-Cysteine 765
sure not exceeding 0.67 kPa, 1009C, 5 hours). solution and the control solution with 4 mL of barium chlo-
ride TS, respectively (not more than 0.028z).
Assay Weigh accurately about 0.5 g of Cyproheptadine L-Cysteine, using the distillation under reduced pressure.
Hydrochloride Hydrate, previously dried, and dissolve in 20 Prepare the control solution with 5.0 mL of Standard Am-
mL of acetic acid (100) by warming at 509C. After cooling, monium Solution (not more than 0.02z).
add 40 mL of acetic anhydride, and titrate <2.50> with 0.1 (5) Heavy metals <1.07>—Proceed with 1.0 g of
mol/L perchloric acid VS (potentiometric titration). Per- L-Cysteine according to Method 4, and perform the test.
form a blank determination, and make any necessary correc- Prepare the control solution with 1.0 mL of Standard Lead
tion. Solution (not more than 10 ppm).
Cysteine according to Method 1, and perform the test ac-
＝ 32.39 mg of C21H21N.HCl
cording to Method A. Prepare the control solution with 1.0
Containers and storage Containers—Well-closed contain- mL of Standard Iron Solution (not more than 10 ppm).
ers. (7) Related substances—Dissolve 0.10 g of L-Cysteine in
N-ethylmaleimide solution (1 in 50) to make exactly 10 mL,
leave for 30 minutes, and use this solution as the sample
L-Cysteine solution. Pipet 1 mL of the sample solution, add water to
make exactly 10 mL, pipet 1 mL of this solution, add water
L-システイン to make exactly 50 mL, and use this solution as the standard
solution (1). Separately, dissolve 0.10 g of L-cystine in 0.5
mol/L hydrochloric acid TS to make exactly 20 mL. Pipet 1
mL of this solution, add water to make 100 mL, and use this
solution as the standard solution (2). Perform the test with
C3H7NO2S: 121.16 these solutions as directed under Thin-layer Chromatogra-
(2R)-2-Amino-3-sulfanylpropanoic acid phy <2.03>. Spot 10 mL each of the sample solution and
[52-90-4] standard solutions (1) and (2) on a plate of silica gel for thin-
L-Cysteine contains not less than 98.5z and not 1-butanol, water, and acetic acid (100) (3:1:1) to a distance
more than 101.0z of L-cysteine (C3H7NO2S), calcu- of about 10 cm, and dry the plate for 30 minutes at 809C.
lated on the dried basis. Spray the plate evenly with a solution of ninhydrin in a mix-
ture of methanol and acetic acid (100) (97:3) (1 in 100), and
Description L-Cysteine occurs as white crystals or a white
then heat at 809C for 10 minutes: the spot obtained from the
crystalline powder. It has a characteristic odor and a pun-
sample solution corresponding to the spot obtained from the
gent taste.
standard solution (2) is not more intense than the spot from
the standard solution (2). Also, the spots other than the prin-
ethanol (99.5).
cipal spot and the spots mentioned above from the sample
Identification Determine the infrared absorption spectrum ard solution (1).
of L-Cysteine as directed in the potassium bromide disk
exhibit similar intensities of absorption at the same wave Residue on ignition <2.44> Not more than 0.1z (1 g).
numbers.
Assay Weigh accurately about 0.2 g of L-Cysteine, place it
Optical rotation <2.49> [a]20D : ＋8.0 – ＋10.09 (2 g calcu- in a stoppered flask, and dissolve in 20 mL of water. Dis-
lated on the dried basis, 1 mol/L hydrochloric acid TS, 25 solve 4 g of potassium iodide in this solution, immediately
mL, 100 mm). place in ice cold water, add 5 mL of dilute hydrochloric acid
and exactly 25 mL of 0.05 mol/L iodine VS, leave in a dark
place for 20 minutes, and then titrate <2.50> an excess
1.25 g of L-Cysteine in 50 mL of water is 4.7 to 5.7.
amount of iodine with 0.1 mol/L sodium thiosulfate VS (in-
Purity (1) Clarity and color of solution—Dissolve 1.0 g dicator: starch TS). Perform a blank determination using the
of L-Cysteine in 20 mL of water: the solution is clear and same method.
colorless.
Each mL of 0.05 mol/L iodine VS ＝ 12.12 mg of C3H7NO2S
(2) Chloride <1.03>—Dissolve 0.30 g of L-Cysteine in 10
mL of diluted nitric acid (1 in 4), add 10 mL of hydrogen Containers and storage Containers—Tight containers.
peroxide (30), heat for 20 minutes in a boiling water bath,
cool, and then add water to make 50 mL. Perform the test
using this solution as the test solution. Prepare the control
solution with 0.35 mL of 0.01 mol/L hydrochloric acid VS
(3) Sulfate <1.14>—Dissolve 0.6 g of L-Cysteine in 30 mL
of water and 3 mL of dilute hydrochloric acid, and add
the test solution. Prepare the control solution as follows: To
0.35 mL of 0.005 mol/L sulfuric acid VS add 3 mL of dilute
hydrochloric acid and water to make 50 mL. Prepare the test
766 L-Cysteine Hydrochloride Hydrate / Official Monographs JP XVII
(6) Related substances—Dissolve 0.10 g of L-Cysteine
L-Cysteine Hydrochloride Hydrate Hydrochloride Hydrate in N-ethylmaleimide solution (1 in
50) to make 10 mL, allow to stand for 30 minutes, and use
L-システイン塩酸塩水和物 this solution as the sample solution. Pipet 1 mL of the sam-
ple solution, add water to make exactly 10 mL. Pipet 1 mL
of this solution, add water to make exactly 50 mL, and use
these solutions as directed under Thin-Layer Chromatogra-
C3H7NO2S.HCl.H2O: 175.63 phy <2.03>. Spot 5 mL each of the sample solution and stand-
(2R)-2-Amino-3-sulfanylpropanoic acid monohydrochloride ard solution on a plate of silica gel for thin-layer chromatog-
monohydrate raphy. Then develop with a mixture of 1-butanol, water and
[7048-04-6] acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry
the plate at 809C for 30 minutes. Spray evenly a solution of
L-Cysteine Hydrochloride Hydrate contains not less ninhydrin in a mixture of methanol and acetic acid (100)
than 98.5z and not more than 101.0z of L-cysteine (97:3) (1 in 100) on the plate, and then heat at 809C for 10
hydrochloride (C3H7NO2S.HCl: 157.62), calculated on minutes: the spot other than the principal spot obtained with
the dried basis. the sample solution is not more intense than the spot ob-
tained with the standard solution.
Description L-Cysteine Hydrochloride Hydrate occurs as
white, crystals or crystalline powder. It has a characteristic Loss on drying <2.41> 8.5 – 12.0z (1 g, in vacuum, phos-
odor and a strong acid taste. phorus (V) oxide, 20 hours).
It is very soluble in water, and soluble in ethanol (99.5).
Assay Weigh accurately about 0.25 g of L-Cysteine Hydro-
chloride Hydrate, place in a glass-stoppered flask, and dis-
trum of L-Cysteine Hydrochloride Hydrate as directed in the
solve in 20 mL of water. Dissolve 4 g of potassium iodide in
potassium chloride disk method under Infrared Spectropho-
this solution, soak immediately in ice cold water, add 5 mL
of dilute hydrochloric acid and exactly 25 mL of 0.05 mol/L
iodine VS, allow to stand for 20 minutes in a dark place,
titrate <2.50> the excess of iodine with 0.1 mol/L sodium
(2) To 10 mL of a solution of L-Cysteine Hydrochloride
thiosulfate VS (indicator: starch TS). Perform a blank deter-
Hydrate (1 in 50) add 1 mL of hydrogen peroxide (30), heat
mination in the same manner, and make any necessary cor-
on a water bath for 20 minutes, and cool: the solution re-
rection.
sponds to the Qualitative Tests <1.09> (2) for chloride.
Each mL of 0.05 mol/L iodine VS
D : ＋6.0 – ＋7.59(2 g, calculated
＝ 15.76 mg of C3H7NO2S.HCl
on the dried basis, 6 mol/L hydrochloric acid TS, 25 mL,
100 mm). Containers and storage Containers—Tight containers.
1.0 g of L-Cysteine Hydrochloride Hydrate in 100 mL of
water is between 1.3 and 2.3. L-Cystine
Purity (1) Clarity and color of solution—A solution ob- L-シスチン
tained by dissolving 1.0 g of L-Cysteine Hydrochloride Hy-
drate in 10 mL of water is clear and colorless.
(2) Sulfate <1.14>—Dissolve 0.8 g of L-Cysteine Hydro-
chloride Hydrate in 30 mL of water and 3 mL of dilute hy-
drochloric acid, and add water to make 50 mL. Perform the
test using this solution as the test solution. Prepare the con- C6H12N2O4S2: 240.30
trol solution as follows: To 0.35 mL of 0.005 mol/L sulfuric 3,3?-Disulfanediylbis[(2R)-2-aminopropanoic acid]
acid VS add 3 mL of dilute hydrochloric acid and water to [56-89-3]
make 50 mL. To both of the test solution and the control
solution add 4 mL of barium chloride TS (not more than L-Cystine, when dried, contains not less than
0.021z). 99.0z and not more than 101.0z of L-cystine
(3) Ammonium <1.02>—Perform the test with 0.25 g of (C6H12N2O4S2).
L-Cysteine Hydrochloride Hydrate using the distillation
Description L-Cystine occurs as white, crystals or crystal-
under reduced pressure. Prepare the control solution with
line powder.
5.0 mL of Standard Ammonium Solution (not more than
0.02z).
L-Cysteine Hydrochloride Hydrate according to Method 4, Identification Determine the infrared absorption spectrum
and perform the test. Prepare the control solution with 1.0 of L-Cystine as directed in the potassium bromide disk
mL of Standard Lead Solution (not more than 10 ppm). method under Infrared Spectrophotometry <2.25>, and com-
(5) Iron <1.10>—Prepare the test solution with 1.0 g of pare the spectrum with the Reference Spectrum: both spectra
L-Cysteine Hydrochloride Hydrate according to Method 1, exhibit similar intensities of absorption at the same wave
and perform the test according to Method A. Prepare the numbers.
control solution with 1.0 mL of Standard Iron Solution (not
D : －215 – －2259(after drying,
more than 10 ppm).
1 g, 1 mol/L hydrochloric acid TS, 50 mL, 100 mm).
JP XVII Official Monographs / Cytarabine 767

tained by dissolving 1.0 g of L-Cystine in 10 mL of 2 mol/L Cytarabine
hydrochloric acid TS is clear and colorless.
(2) Chloride <1.03>—Dissolve 0.5 g of L-Cystine in 10 シタラビン
mL of dilute nitric acid, add 10 mL of hydrogen peroxide
(30), and heat in a water bath for 10 minutes. After cooling,
add water to make 50 mL, and perform the test using this so-
lution as the test solution. Prepare the control solution with
0.021z).
(3) Sulfate <1.14>—Dissolve 0.6 g of L-Cystine in 5 mL
of dilute hydrochloric acid, add water to make 45 mL, and
pare the control solution as follows: To 0.35 mL of 0.005 C9H13N3O5: 243.22
mol/L sulfuric acid VS add 5 mL of dilute hydrochloric acid 1-b-D-Arabinofuranosylcytosine
and water to make 45 mL. To both the test and control solu- [147-94-4]
tions add 5 mL of barium chloride TS (not more than
0.028z). Cytarabine, when dried, contains not less than
(4) Ammonium <1.02>—Perform the test with 0.25 g of 98.5z and not more than 101.0z of cytarabine
L-Cystine, using the distillation under reduced pressure. Pre- (C9H13N3O5).
pare the control solution with 5.0 mL of Standard Ammoni-
Description Cytarabine occurs as white, crystals or crystal-
um Solution (not more than 0.02z).
line powder.
It is freely soluble in water, soluble in acetic acid (100),
L-Cystine according to Method 4, and perform the test.
and very slightly soluble in ethanol (99.5).
Cystine according to Method 3, and perform the test accord- Identification (1) Determine the absorption spectrum of a
ing to Method A. Prepare the control solution with 1.0 mL solution of Cytarabine in 0.1 mol/L hydrochloric acid TS
of Standard Iron Solution (not more than 10 ppm). (1 in 100,000) as directed under Ultraviolet-visible Spectro-
(7) Related substances—Dissolve 0.20 g of L-Cystine in photometry <2.24>, and compare the spectrum with the Ref-
20 mL of 1 mol/L hydrochloric acid TS, and use this solu- erence Spectrum: both spectra exhibit similar intensities of
tion as the sample solution. Pipet 1 mL of the sample solu- absorption at the same wavelengths.
tion, and add water to make exactly 10 mL. Pipet 1 mL of (2) Determine the infrared absorption spectrum of
this solution, add water to make exactly 50 mL, and use this Cytarabine as directed in the potassium bromide disk
solution as the standard solution. Perform the test with these method under Infrared Spectrophotometry <2.25>, and com-
solutions as directed under Thin-layer Chromatography pare the spectrum with the Reference Spectrum: both spectra
<2.03>. Spot 5 mL each of the sample solution and standard exhibit similar intensities of absorption at the same wave
solution on a plate of silica gel for thin-layer chromatogra- numbers.
phy. Develop the plate with a mixture of 1-propanol and am-
monia solution (28) (67:33) to a distance of about 10 cm, and
0.1 g, water, 10 mL, 100 mm).
dry the plate at 809 C for 30 minutes. Spray evenly a solution
of ninhydrin in a mixture of methanol and acetic acid (100) pH <2.54> Dissolve 0.20 g of Cytarabine in 20 mL of water:
(97:3) (1 in 100) on the plate, and heat at 809C for 10 the pH of this solution is between 6.5 and 8.0.
minutes: the spot other than the principal spot obtained
from the sample solution is not more intense than the spot
of Cytarabine in 10 mL of water: the solution is clear and
obtained from the standard solution.
colorless.
Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, (2) Chloride <1.03>—Perform the test with 1.0 g of
3 hours). Cytarabine. Prepare the control solution with 0.25 mL of
(3) Heavy metals <1.07>—Proceed with 1.0 g of Cytara-
Assay Weigh accurately about 30 mg of L-Cystine, previ- bine according to Method 1, and perform the test. Prepare
ously dried, and perform the test as directed under Nitrogen the control solution with 2.0 mL of Standard Lead Solution
Determination <1.08>. (not more than 20 ppm).
(4) Related substances—Dissolve 0.10 g of Cytarabine in
10 mL of water, and use this solution as the sample solution.
＝ 1.202 mg of C6H12N2O4S2
Pipet 1 mL of the sample solution, add water to make ex-
Containers and storage Containers—Tight containers. actly 200 mL, and use this solution as the standard solution
Storage—Light-resistant. (1). Pipet 10 mL of the standard solution (1), add water to
make exactly 25 mL and use this solution as the standard so-
lution (2). Perform the test with these solutions as directed
of the sample solution and standard solutions (1) and (2) on
a plate of silica gel with fluorescent indicator for thin-layer
chromatography. Develop the plate with 1-butanol saturated
with water to a distance of about 12 cm, and air-dry the
768 Danazol / Official Monographs JP XVII
plate. Examine under ultraviolet light (main wavelength: 254 tion as the test solution. Prepare the control solution with
nm): the spots other than the principal spot from the sample 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than
solution are not more intense than the spot from the stand- 0.011z).
ard solution (1), and the number of them which are more in- (2) Heavy metals <1.07>—Proceed with 2.0 g of Danazol
tense than the spot from the standard solution (2) is not according to Method 2, and perform the test. Prepare the
more than two. Spray evenly acidic potassium permanganate control solution with 2.0 mL of Standard Lead Solution (not
TS on the plate: any spot other than the principal spot does more than 10 ppm).
not appear. (3) Related substances—Dissolve 0.20 g of Danazol in 4
mL of acetone, and use this solution as the sample solution.
Pipet 2 mL of the sample solution, add acetone to make ex-
actly 200 mL. Pipet 4 mL of this solution, add acetone to
Residue on ignition <2.44> Not more than 0.5z (1 g). make exactly 20 mL, and use this solution as the standard
Assay Weigh accurately about 0.2 g of Cytarabine, previ-
under Thin-layer Chromatography <2.03>. Spot 5 mL each of
gel with fluorescent indicator for thin-layer chromatogra-
phy. Develop the plate with a mixture of cyclohexane and
ethyl acetate (3:2) to a distance of about 15 cm, and air-dry
Each mL of 0.05 mol/L perchloric acid VS the plate. Examine under ultraviolet light (main wavelength:
＝ 12.16 mg of C9H13N3O5 254 nm): the spots other than the principal spot and the spot
of the starting point obtained from the sample solution are
not more intense than the spot obtained from the standard
solution.
Danazol Loss on drying <2.41> Not more than 0.2z (1 g, in vacu-

um, phosphorous (V) oxide, 609
C, 4 hours).
ダナゾール
Assay Weigh accurately about 25 mg each of Danazol and
Danazol RS, previously dried, dissolve separately in ethanol
(95) to make exactly 50 mL. Pipet 2 mL each of these solu-
tions, add ethanol (95) to make exactly 50 mL, and use these
solutions as the sample solution and the standard solution,
respectively. Perform the test with the sample solution and
C22H27NO2: 337.46
17a-Pregna-2,4-dien-20-yno[2,3-d ]isoxazol-17-ol
trophotometry <2.24>, and determine the absorbances, AT
[17230-88-5]
and AS, at 285 nm.
Danazol, when dried, contains not less than 98.5z Amount (mg) of danazol (C22H27NO2)
and not more than 101.0z of danazol (C22H27NO2). ＝ MS × AT/AS
Description Danazol occurs as a white to pale yellow crys- MS: Amount (mg) of Danazol RS taken
talline powder.
It is soluble in acetone, sparingly soluble in ethanol (99.5),
ers.
and practically insoluble in water.
solution of Danazol in ethanol (95) (1 in 50,000) as directed Dantrolene Sodium Hydrate
compare the spectrum with the Reference Spectrum or the ダントロレンナトリウム水和物
spectrum of a solution of Danazol RS prepared in the same
Danazol as directed in the potassium bromide disk method
spectrum with the Reference Spectrum or the spectrum of C14H9N4NaO5.3 1/2 H2O: 399.29
Danazol RS: both spectra exhibit similar intensities of ab- Monosodium 3-[5-(4-nitrophenyl)furan-
sorption at the same wave numbers. 2-ylmethylene]amino-2,5-dioxo-1,3-imidazolidinate
hemiheptahydrate
Optical rotation <2.49> [a]20D : ＋8 – ＋119 (after drying,
[14663-23-1, anhydride]
0.25 g, ethanol (99.5), 50 mL, 100 mm).
Purity (1) Chloride <1.03>—To 2.0 g of Danazol add 80 Dantrolene Sodium Hydrate contains not less than
mL of water, shake well, and boil for 5 minutes. After cool- 98.0z of dantrolene sodium (C14H9N4NaO5: 336.23),
ing, add water to make 100 mL, and filter through a glass calculated on the anhydrous basis.
filter (G4). Discard the first 30 mL of the filtrate, take 40 mL
Description Dantrolene Sodium Hydrate occurs as a yel-
of the subsequent filtrate, and add 6 mL of dilute nitric acid
lowish orange to deep orange, crystalline powder.
and water to make 50 mL. Perform the test using this solu-

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What are the identification tests for Acebutolol Hydrochloride?

What are the identification tests for Acebutolol Hydrochloride?

What are the purity tests for Acebutolol Hydrochloride?

What are the purity tests for Acebutolol Hydrochloride?

Official Monographs

water, 1-butanol and acetic acid (100) (5:4:1) to a distance of

Residue on ignition <2.44> Not more than 0.1z (1 g).

Tablets is not less than 80z. mg of indometacin in 50 mL of methanol. To 4 mL of this

Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C,

Column: A stainless steel column 4.6 mm in inside diame-

2.0z. more than 10 ppm).

Perform the test with exactly 20 mL each of the sample solu-

L-Alanine. Prepare the control solution with 5.0 mL of

tions per minute according to the Paddle method, using 900

silanized silica gel for liquid chromatography (5 mm in parti-

Dried Aluminum Potassium Sulfate

Natural Aluminum Silicate

Description Natural Aluminum Silicate occurs as a white

System repeatability: When the test is repeated 6 times

Foreign insoluble matter <6.06> Perform the test according

Amitriptyline Hydrochloride Tablets contain not

50 mL. Pipet 25 mL of this solution, add exactly 10 mL of

hydrous basis. peak.

Amoxapine, when dried, contains not less than

suitability test under the above operating conditions, N, N-

Amyl Nitrite Dental Antiformin

Antipyrine Aprindine Hydrochloride

to Method 1: it meets the requirement. Optical rotation <2.49> [a]20

Argatroban Hydrate contains not less than 0–5 100 0

allow to stand in a dark place at room temperature for 45

these solutions as directed under Thin-layer Chromatogra-

pare the control solution with 2.0 mL of Standard Lead So-

area of azelastine is not more than 2.0z.

Freeze-dried BCG Vaccine (for Percutaneous Use) is

drops of alizarin red S TS. 354.01), calculated on the anhydrous basis.

Benzalkonium Chloride Assay Weigh accurately about 0.15 g of Benzalkonium

standard solution. Perform the test with these solutions as

Description Benzylpenicillin Benzathine Hydrate occurs as 0 – 10 75 25

sodium (C24H29NaO5). peak of beraprost is about 23 minutes.

with 20 mL of the standard solution under the above operat-

these solutions as directed under Thin-layer Chromatogra- suitability test.

Assay Weigh accurately about 0.4 g of Bethanechol Chlo-

(5) Dissolve 0.02 g of Biperiden Hydrochloride in 10 mL tically insoluble in water.

of nitric acid by warming. Pour the solution into 100 mL of Storage—Light-resistant.

System repeatability: When the test is repeated 6 times

the dried basis. The potency of Bleomycin Hydrochlo-

Description Bleomycin Hydrochloride occurs as a white to 0 – 60 100 → 0 0 → 100

Bleomycin Sulfate is the sulfate of a mixture of sub-

is expressed as mass (potency) of bleomycin A2

Boric Acid, when dried, contains not less than

condenser and the mouth of the flask with 30 mL of water,

Bromovalerylurea, when dried, contains not less

gel for liquid chromatography (5 mm in particle diameter).

Busulfan Butenafine Hydrochloride

taken as directed under Ultraviolet-visible Spectrophotometry

with 10 mL of the standard solution under the above operat-

length: 272 nm). Melting point: about 1659C (with decomposition).

System suitability— (2) Sulfate <1.14>—To 40 mL of the sample solution ob-

Time after Mobile Mobile Mobile Mobile Mobile Operating conditions—