EUROPEAN PHARMACOPOEIA 11.

2 Ascorbic acid

07/2023:0253 Allow the solutions to stand for 1 h. Any opalescence in the
test solution is not more intense than that in the reference
solution.
Related substances. Liquid chromatography (2.2.29). Prepare
the solutions immediately before use.
ASCORBIC ACID Phosphate buffer solution. Dissolve 6.8 g of potassium
dihydrogen phosphate R in water for chromatography R and
dilute to about 200 mL with the same solvent. Filter through
Acidum ascorbicum a membrane filter (nominal pore size 0.45 μm) and dilute to
1000 mL with water for chromatography R.
Test solution. Dissolve 0.500 g of the substance to be examined
in the mobile phase and dilute to 10.0 mL with the mobile
phase.
Reference solution (a). Dissolve 10.0 mg of ascorbic acid
impurity C CRS in the mobile phase and dilute to 5.0 mL with
C 6H 8O 6 Mr 176.1 the mobile phase.
[50-81-7] Reference solution (b). Dissolve 5.0 mg of ascorbic acid
impurity D CRS and 5.0 mg of ascorbic acid CRS in the mobile
DEFINITION phase and dilute to 100.0 mL with the mobile phase.
(5R)-5-[(1S)-1,2-Dihydroxyethyl]-3,4-dihydroxyfuran-2(5H)- Reference solution (c). Dilute 1 mL of the test solution to
one. 200 mL with the mobile phase. Mix 1 mL of this solution and
Content : 99.0 per cent to 100.5 per cent. 1 mL of reference solution (a).
CHARACTERS Reference solution (d). Dilute 2.5 mL of reference solution (a)
to 100.0 mL with the mobile phase.
Appearance : white or almost white, crystalline powder or
colourless crystals, becoming discoloured on exposure to air Column :
and moisture. – size : l = 0.25 m, Ø = 4.6 mm ;
Solubility : freely soluble in water, sparingly soluble in ethanol – stationary phase : aminopropylsilyl silica gel for
(96 per cent). chromatography R (5 μm) ;
mp : about 190 °C, with decomposition. – temperature : 45 °C.
Mobile phase : phosphate buffer solution, acetonitrile R1
IDENTIFICATION (25:75 V/V).
First identification : B, C. Flow rate : 1.0 mL/min.
Second identification : A, C, D. Detection : spectrophotometer at 210 nm.
A. Ultraviolet and visible absorption spectrophotometry Injection : 20 μL of the test solution and reference solutions (b),
(2.2.25). (c) and (d).
Test solution. Dissolve 0.10 g in water R and dilute Run time : 2.5 times the retention time of ascorbic acid.
immediately to 100.0 mL with the same solvent. Add Identification of impurities: use the chromatogram obtained
1.0 mL of the solution to 10 mL of a 10.3 g/L solution of with reference solution (d) to identify the peak due to
hydrochloric acid R and dilute to 100.0 mL with water R. impurity C ; use the chromatogram obtained with reference
Absorption maximum : 243 nm, determined immediately solution (b) to identify the peak due to impurity D.
after dissolution. Relative retention with reference to ascorbic acid
Specific absorbance at the absorption maximum : 545 to 585. (retention time = about 9 min) : impurity D = about 0.5 ;
B. Infrared absorption spectrophotometry (2.2.24). impurity C = about 1.45.
Comparison : ascorbic acid CRS. System suitability :
C. pH (2.2.3) : 2.1 to 2.6 for solution S (see Tests). – resolution : minimum 3.0 between the peaks due to ascorbic
acid and impurity C in the chromatogram obtained with
D. To 1 mL of solution S add 0.2 mL of dilute nitric acid R reference solution (c) ;
and 0.2 mL of silver nitrate solution R2. A grey precipitate
– signal-to-noise ratio : minimum 20 for the peak due to
is formed.
impurity C in the chromatogram obtained with reference
TESTS solution (d).
Solution S. Dissolve 1.0 g in carbon dioxide-free water R and Calculation of percentage contents :
dilute to 20 mL with the same solvent. – for impurity C, use the concentration of impurity C in
reference solution (d) ;
Appearance of solution. Solution S is clear (2.2.1) and not
more intensely coloured than reference solution BY7 (2.2.2, – for impurity D, use the concentration of impurity D in
Method II). reference solution (b);
– for impurities other than C and D, use the concentration of
Specific optical rotation (2.2.7) : + 20.5 to + 21.5.
ascorbic acid in reference solution (b).
Dissolve 2.50 g in water R and dilute to 25.0 mL with the same
Limits :
solvent.
– impurities C, D : for each impurity, maximum 0.15 per cent ;
Impurity E : maximum 0.2 per cent.
– unspecified impurities : for each impurity, maximum
Test solution. Dissolve 0.25 g in 5 mL of water R. Neutralise 0.10 per cent ;
using dilute sodium hydroxide solution R, then add 1 mL of
– sum of impurities other than C and D : maximum 0.2 per
dilute acetic acid R and 0.5 mL of calcium chloride solution R.
cent ;
Reference solution. Dissolve 70 mg of oxalic acid R (dihydrate – reporting threshold : 0.05 per cent.
of impurity E) in water R and dilute to 500 mL with the same
solvent ; to 5 mL of the solution add 1 mL of dilute acetic Copper : maximum 5 ppm.
acid R and 0.5 mL of calcium chloride solution R. Atomic absorption spectrometry (2.2.23, Method I).

General Notices (1) apply to all monographs and other texts 4865
Ascorbic acid EUROPEAN PHARMACOPOEIA 11.2

Test solution. Dissolve 2.0 g in 0.1 M nitric acid and dilute to

25.0 mL with the same acid.
Reference solutions. Prepare the reference solutions (0.2 ppm,
0.4 ppm and 0.6 ppm) using copper standard solution (10 ppm A. furan-2-carbaldehyde (furfural),
Cu) R, diluting with 0.1 M nitric acid.
Source : copper hollow-cathode lamp.
Wavelength : 324.8 nm.
Atomisation device : air-acetylene flame.
Adjust the zero of the apparatus using 0.1 M nitric acid. C. L-xylo-hex-2-ulosonic acid (L-sorbosonic acid),
Iron : maximum 2 ppm.
Atomic absorption spectrometry (2.2.23).
Test solution. Dissolve 5.0 g in 0.1 M nitric acid and dilute to
25.0 mL with the same acid.
Reference solutions. Prepare the reference solutions (0.2 ppm,
0.4 ppm and 0.6 ppm) using iron standard solution (20 ppm
Fe) R, diluting with 0.1 M nitric acid. D. methyl L-xylo-hex-2-ulosonate (methyl L-sorbosonate),
Source : iron hollow-cathode lamp.
Wavelength : 248.3 nm.
E. oxalic acid,
Atomisation device : air-acetylene flame.
Adjust the zero of the apparatus using 0.1 M nitric acid.
Sulfated ash (2.4.14) : maximum 0.1 per cent, determined on
1.0 g.
ASSAY
Dissolve 0.150 g in a mixture of 10 mL of dilute sulfuric F. (5R)-5-[(1R)-1,2-dihydroxyethyl]-3,4-dihydroxyfuran-
acid R and 80 mL of carbon dioxide-free water R. Add 1 mL of 2(5H)-one,
starch solution R. Titrate with 0.05 M iodine until a persistent
violet-blue colour is obtained.
1 mL of 0.05 M iodine is equivalent to 8.81 mg of C6H8O6.
STORAGE
In a non-metallic container, protected from light.
G. (R)-[(2R)-3,4-dihydroxy-5-oxo-2,5-dihydrofuran-2-
IMPURITIES yl]hydroxyacetic acid,
Specified impurities : C, D, E.
Other detectable impurities (the following substances would,
if present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these impurities
for demonstration of compliance. See also 5.10. Control of H. methyl (R)-[(2R)-3,4-dihydroxy-5-oxo-2,5-dihydrofuran-
impurities in substances for pharmaceutical use): A, F, G, H. 2-yl]hydroxyacetate.

4866 See the information section on general monographs (cover pages)