Printed on: Wed Mar 29 2023, 08:17:02 PM(EST) Status: Currently Official on 30-Mar-2023 DocId: GUID-49589C56-503E-4019-B930-9DC45304D336_5_en-US

Printed by: Maria Christina Official Date: Official as of 01-Oct-2021 Document Type: USP @2023 USPC
Do Not Distribute DOI Ref: cce10 DOI: https://doi.org/10.31003/USPNF_M58350_05_01
1

System suitability
Ofloxacin Sample: System suitability solution
Suitability requirements
Change to read: Resolution: NLT 2.0 between ofloxacin and ofloxacin
related compound A
Relative standard deviation: NMT 3.0%
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of each impurity in the portion
of Ofloxacin taken:
C18H20FN3O4 361.37
7H-Pyrido[1,2,3-de]-1,4-benzoxazine-6-carboxylic acid, 9- Result = (r U/r S) × (C S/C U) × 100
fluoro-2,3-dihydro-3-methyl-10-(4-methyl-1-piperazinyl)-7-
oxo-, (±)-; rU = peak response from the Sample solution
(±)-9-Fluoro-2,3-dihydro-3-methyl-10-(4-methyl- rS = peak response of ofloxacin from the Standard
▲
1▲ (ERR 1-Oct-2021)-piperazinyl)-7-oxo-7H-pyrido[1,2,3-de]- solution
1,4-benzoxazine-6-carboxylic acid CAS RN®: 82419-36-1; CS = concentration of USP Ofloxacin RS in the Standard
UNII: A4P49JAZ9H. solution (µg/mL)
CU = concentration of the Sample solution (µg/mL)
DEFINITION
Ofloxacin contains NLT 98.5% and NMT 101.5% of ofloxacin Acceptance criteria
(C18H20FN3O4), calculated on the dried basis. Individual impurities: NMT 0.3%. Disregard any peak less
than 0.02%.

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IDENTIFICATION
Total impurities: NMT 0.5%
• A. SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared
• LIMIT OF METHANOL AND ETHANOL
Spectroscopy: 197K
Internal standard solution: 5.6 nL/mL of n-propyl alcohol
• B. SPECTROSCOPIC IDENTIFICATION TESTS á197ñ,
in sodium hydroxide solution (1 in 100)
Ultraviolet-Visible Spectroscopy: 197U
Standard solution: 10.0 µg/mL each of methanol and
Medium: 0.1 N hydrochloric acid
Sample solution: 6.7 µg/mL in Medium
Acceptance criteria: Meets the requirements
ci dehydrated alcohol in Internal standard solution. Transfer
2 mL of this solution to a vial fitted with a septum and crimp
cap, and seal. Heat the sealed vial at 90° for 2 min, and
ASSAY shake for 6 min.
• PROCEDURE Sample solution: Transfer 40 mg of Ofloxacin to a vial fitted
ffi
Sample: 100 mg of Ofloxacin with a septum and crimp cap, add 2 mL of Internal standard
Analysis: Dissolve the Sample in 275 mL of acetic anhydride. solution, and seal the vial. Heat the sealed vial at 90° for
Titrate with 0.1 N perchloric acid VS, determining the 2 min, and shake for 6 min.
endpoint potentiometrically, using a glass–silver chloride Blank: Transfer 2 mL of Internal standard solution to a vial
electrode system (see Titrimetry á541ñ). Use the first of the fitted with a septum and crimp cap, and seal. Heat the
two inflection points. Perform a blank determination, and sealed vial at 90° for 2 min, and shake for 6 min.
O

make any necessary correction. Each mL of 0.1 N perchloric Chromatographic system

acid is equivalent to 36.138 mg of C18H20FN3O4. (See Chromatography á621ñ, System Suitability.)
Acceptance criteria: 98.5%–101.5% on the dried basis Mode: GC
Detector: Flame ionization
IMPURITIES Column: 0.53-mm × 30-m fused silica column, coated
• RESIDUE ON IGNITION á281ñ: NMT 0.1% with a 3.0-µm film of stationary phase G43, and a fused
• ARSENIC, Method II á211ñ: NMT 1 µg/g silica precolumn
• ORGANIC IMPURITIES Temperatures
Diluent: Acetonitrile and water (1:6) Injector: 170°
Buffer: 3.1 g/L of ammonium acetate and 5.4 g/L of sodium Detector: 250°
perchlorate in water. Adjust with phosphoric acid to a pH Column: See Table 1.
of 2.2.
Mobile phase: Acetonitrile and Buffer (240:1300) Table 1
System suitability solution: 0.4 µg/mL each of USP Hold Time at Fi-
Ofloxacin RS and USP Ofloxacin Related Compound A RS Initial Temperature Final nal
in Diluent Temperature Ramp Temperature Temperature
Standard solution: 0.4 µg/mL of USP Ofloxacin RS in (°) (°/min) (°) (min)
Diluent 35 — 35 3
Sample solution: 0.2 mg/mL of Ofloxacin in Diluent
Chromatographic system 35 20 90 —
(See Chromatography á621ñ, System Suitability.) 90 40 200 2
Mode: LC
Detector: UV 294 nm
Column: 4.6-mm × 15-cm; packing L1 Condition the column with the helium flowing at 200°
Column temperature: 45° for 2 h or until a stable baseline is obtained.
Flow rate: 0.5 mL/min Carrier gas: Helium
Injection volume: 10 µL Flow rate: 7 mL/min
Run time: 2.5 times the retention time of the Injection volume: 1-mL headspace. Use a heated,
ofloxacin peak gas-tight syringe to make injections of the headspace into
the chromatograph.

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Printed on: Wed Mar 29 2023, 08:17:02 PM(EST) Status: Currently Official on 30-Mar-2023 DocId: GUID-49589C56-503E-4019-B930-9DC45304D336_5_en-US
Printed by: Maria Christina Official Date: Official as of 01-Oct-2021 Document Type: USP @2023 USPC
Do Not Distribute DOI Ref: cce10 DOI: https://doi.org/10.31003/USPNF_M58350_05_01
2

System suitability CU = concentration of Ofloxacin in the Sample solution

Sample: Standard solution (µg/mL)
[NOTE—The relative retention times for methanol,
ethanol, and n-propyl alcohol are 0.5, 0.6, and 1.0, Acceptance criteria: NMT 0.005% of methanol and NMT
respectively.] 0.05% of ethanol
Suitability requirements
Resolution: NLT 2.0 between methanol and ethanol SPECIFIC TESTS
Relative standard deviation: NMT 5% • OPTICAL ROTATION, Specific Rotation á781Sñ
Analysis Sample solution: 10 mg/mL in chloroform
Samples: Standard solution, Sample solution, and Blank Acceptance criteria: +1° to −1°
Calculate the percentage of methanol and ethanol in the • LOSS ON DRYING á731ñ
portion of Ofloxacin taken: Analysis: Dry at 105° for 4 h.
Acceptance criteria: NMT 0.2%
Result = (R U/R S ) × (C S/C U) × 100 ADDITIONAL REQUIREMENTS
• PACKAGING AND STORAGE: Preserve in well-closed
RU = peak response ratio of methanol or ethanol to the containers, protected from light. Store at 25°, excursions
internal standard, corrected for the blank, from permitted between 15° and 30°.
the Sample solution • USP REFERENCE STANDARDS á11ñ
RS = peak response ratio of methanol or ethanol to the USP Ofloxacin RS
internal standard, corrected for the blank, from USP Ofloxacin Related Compound A RS
the Standard solution 9-Fluoro-3-methyl-7-oxo-10-(piperazin-1-yl)-2,3-
CS = concentration of methanol or ethanol in the dihydro-7H-pyrido[1,2,3-de]-1,4-benzoxazine-6-
Standard solution (µg/mL)

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carboxylic acid.
C17H18FN3O4 347.34

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ffi
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