3432Ciprofloxacin / Official MonographsUSP 40IDENTIFICATION•USP R EFERENCE S TANDARDS 〈11〉• The retention time of the major peak of the Sample solu-USP Ciprofloxacin Ethylenediamine Analog RStion corresponds to that of the Standard solution , as ob-1-Cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-[(2-ami-tained in the Assay .noethyl)amino]-3-quinolinecarboxylic acid hydrochloride.ASSAYC 15H 16FN 3O 3·HCl 341.77•P ROCEDUREUSP Ciprofloxacin Hydrochloride RSSolution A: 0.005 M tetrabutylammonium phosphate solution. Adjust with phosphoric acid to a pH of 2.0.Mobile phase: Methanol and Solution A (1:3)Standard solution: 0.14mg/mL of USP Ciprofloxacin Hydrochloride RS in waterCiprofloxacin TabletsSystem suitability solution: 0.01mg/mL of USP Ciprofloxacin Ethylenediamine Analog RS in Standard DEFINITIONsolutionCiprofloxacin Tablets contain Ciprofloxacin Hydrochloride Sample solution: Equivalent to 0.12mg/mL of equivalent to NLT 90.0% and NMT 110.0% of the labeled ciprofloxacin from Ophthalmic Solution, in water amount of ciprofloxacin (C 17H 18FN 3O 3).Chromatographic system(See Chromatography 〈621〉, System Suitability .)IDENTIFICATIONMode: LCThe retention time of the major peak of the Sample solution Detector: UV 280 nmcorresponds to that of the Standard solution , as obtained Column: 4.6-mm × 25-cm; packing L1in the Assay .Flow rate: 1.5mL/min Injection size: 20µL ASSAYSystem suitability•P ROCEDURESamples: Standard solution and System suitability Solution A: 0.025 M phosphoric acid. Adjust with tri-solutionethylamine to a pH of 2.0 ± 0.1.[N OTE —The relative retention times for the ciprofloxacin Solution B: Acetonitrile and Solution A (13:87)ethylenediamine analog and ciprofloxacin are 0.8 and Solution C: 0.025 M phosphoric acid. Adjust with tri-1.0, respectively.]ethylamine to a pH of 3.0 ± 0.1.Suitability requirementsMobile phase: Acetonitrile and Solution C (13:87)Resolution: NLT 1.5 between the ciprofloxacin ethyl-Standard solution: 0.2mg/mL of USP Ciprofloxacin enediamine analog and ciprofloxacin, System suitabil-Hydrochloride RS in Solution Bity solutionSystem suitability solution: 0.05mg/mL of USPCapacity factor: 1.5–6 for the ciprofloxacin peak,Ciprofloxacin Ethylenediamine Analog RS in the Stan-Standard solutiondard solutionColumn efficiency: NLT 500 theoretical plates, Stan-Sample solution: Transfer 5 Tablets to a 500-mL volu-dard solutionmetric flask, add 400mL of Solution B , and sonicate for Tailing factor: 0.9–2.0, Standard solutionabout 20 min. Dilute with Solution B to volume, mix,Relative standard deviation: NMT 2%, Standard and pass through a membrane filter of 0.45-µm pore solution size. Prepare the equivalent of 0.20mg/mL of ciproflox-Analysisacin from the filtrate with Solution B .Samples: Standard solution and Sample solution Chromatographic systemCalculate the percentage of the labeled amount of (See Chromatography 〈621〉, System Suitability .)C 17H 18FN 3O 3 in the portion of Ophthalmic Solution Mode: LCtaken:Detector: UV 278 nmColumn: 4.6-mm × 25-cm; packing L1Result = (r U /r S ) × (C S /C U ) × (M r1/M r2) × 100Column temperature: 30±1°Flow rate: 1.5mL/min r U = peak response from the Sample solution Injection size: 10µL r S = peak response from the Standard solution System suitabilityC S= concentration of USP CiprofloxacinSamples: Standard solution and System suitability Hydrochloride RS in the Standard solution solution(mg/mL)[N OTE —The retention time for ciprofloxacin is 6.4–10.8C U = nominal concentration of ciprofloxacin in themin. The relative retention times for ciprofloxacin eth-Sample solution (mg/mL)ylenediamine analog and ciprofloxacin are 0.7 and M r1= molecular weight of ciprofloxacin, 331.34 1.0, respectively.]M r2= molecular weight of anhydrous ciprofloxacinSuitability requirementshydrochloride, 367.81Resolution: NLT 6 between the ciprofloxacin ethyl-Acceptance criteria: 90.0%–110.0%enediamine analog peak and the ciprofloxacin peak,System suitability requirementsSPECIFIC TESTS Column efficiency: NLT 2500 theoretical plates from •P H 〈791〉: 3.5–5.5the ciprofloxacin peak, Standard solution•S TERILITY T ESTS 〈71〉: It meets the requirements when Tailing factor: NMT 2.0 for the ciprofloxacin peak,tested as directed under Test for Sterility of the Product to Standard solutionBe Examined, Membrane Filtration .Relative standard deviation: NMT 1.5%, Standard solution ADDITIONAL REQUIREMENTSAnalysis•P ACKAGING AND S TORAGE : Preserve in tight containers,Samples: Standard solution and Sample solutionprotected from light, at room temperature.Calculate the percentage of C 17H 18FN 3O 3 in the portion of Tablets taken:Result = (r U /r S ) × (C S /C U ) × (M r1/M r2) × 100r U= peak response from the Sample solutionUSP 40Official Monographs / Ciprofloxacin3433r S= peak response from the Standard solution Sample stock solution: Nominally 0.5mg/mL in MobileC S= concentration of USP Ciprofloxacin phase prepared as follows. Transfer an equivalent toHydrochloride RS in the Standard solution250mg of ciprofloxacin from finely powdered Tablets(mg/mL), calculated on the anhydrous basis(NLT 20) to a 500-mL volumetric flask. Add 400mL ofC U= nominal concentration of ciprofloxacin in the Mobile phase, place on a rotary shaker for 15 min, andSample solution (mg/mL)sonicate for 25 min. Allow the solution to cool to room M r1= molecular weight of ciprofloxacin, 331.34temperature, and dilute with Mobile phase to volume.M r2= molecular weight of anhydrous ciprofloxacin Pass a portion of the solution through a suitable filter of hydrochloride, 367.810.45-µm pore size.Acceptance criteria: 90.0%–110.0%Sample solution: Nominally 0.05mg/mL of ciproflox-acin in water from Sample stock solution PERFORMANCE TESTS Chromatographic system•D ISSOLUTION〈711〉(See Chromatography 〈621〉, System Suitability.) Medium: 0.01 N hydrochloric acid; 900mL Mode: LCApparatus 2: 50 rpm Detector: UV 278 nmTime: 30 min Column: 4.6-mm × 25-cm; 5-µm packing L1Sample solution: Pass a portion of the solution under Column temperature: 30°test through a suitable filter. Dilute with Medium, if Flow rate: 1.5mL/minnecessary.Injection volume: 10µLStandard solution: USP Ciprofloxacin Hydrochloride RS System suitabilityin Medium Samples:System suitability solution and Standard Spectrometric conditions solution(See Ultraviolet-Visible Spectroscopy 〈857〉.)Suitability requirementsMode: UV Resolution: NLT 6 between the ciprofloxacin and Analytical wavelength: 276 nm ciprofloxacin ethylenediamine analog peaks, System Analysis suitability solutionSamples:Sample solution and Standard solution Tailing factor: NMT 4.0 for the ciprofloxacin peak, Tolerances: An amount of ciprofloxacin hydrochloride System suitability solution(C17H18FN3O3·HCl) equivalent to NLT 80% (Q) of the Relative standard deviation: NMT 2.0% for the labeled amount of ciprofloxacin (C17H18FN3O3) is ciprofloxacin peak, Standard solutiondissolved.Analysis•U NIFORMITY OF D OSAGE U NITS〈905〉: Meet the Samples:Standard solution and Sample solution requirements Calculate the percentage of the labeled amount ofciprofloxacin (C17H18FN3O3) in the portion of Tablets ADDITIONAL REQUIREMENTS taken:•P ACKAGING AND S TORAGE: Preserve in well-closedcontainers.Result = (rU/r S) × (C S/C U) × (M r1/M r2) × 100•USP R EFERENCE S TANDARDS〈11〉USP Ciprofloxacin Ethylenediamine Analog RS rU= peak response of ciprofloxacin from the 1-Cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-[(2-ami-Sample solutionnoethyl)amino]-3-quinolinecarboxylic acid rS= peak response of ciprofloxacin from the hydrochloride.Standard solutionC15H16FN3O3·HCl341.77CS= concentration of USP Ciprofloxacin USP Ciprofloxacin Hydrochloride RS Hydrochloride RS in the Standard solution(mg/mL)C U= nominal concentration of ciprofloxacin in theSample solution (mg/mL)M r1= molecular weight of ciprofloxacin, 331.34 Ciprofloxacin Extended-Release Tablets M r2= molecular weight of anhydrous ciprofloxacinhydrochloride, 367.81Acceptance criteria: 90.0%–110.0%DEFINITIONCiprofloxacin Extended-Release Tablets contain NLT 90.0%PERFORMANCE TESTSand NMT 110.0% of the labeled amount of ciprofloxacin•D ISSOLUTION〈711〉(C17H18FN3O3).Test 1Medium: pH 4.5 acetate buffer (transfer 3g of sodium IDENTIFICATIONacetate and 14mL of 2N acetic acid to a 1-L volumet-•A. The retention time of the major peak of the Sampleric flask, and dilute with water to volume); 900mL, solution corresponds to that of the Standard solution, asdeaeratedobtained in the Assay.Apparatus 2: 50 rpmASSAY Times: 30, 60, and 120 min•P ROCEDURE Standard solution: 6.5µg/mL of USP Ciprofloxacin Buffer: Dilute 2.9mL of phosphoric acid in water to Hydrochloride RS in Medium1000mL. Adjust with triethylamine to a pH of 3.0.Sample solution: Pass a portion of the solution under Mobile phase: Acetonitrile and Buffer (135:865)test through a suitable filter of 0.45-µm pore size. For System suitability solution: 0.58mg/mL of USP500-mg Tablets, transfer 2mL of the filtrate to a Ciprofloxacin Hydrochloride RS and 0.5mg/mL of USP200-mL volumetric flask, and dilute with Medium to Ciprofloxacin Ethylenediamine Analog RS in Mobile volume. For 1000-mg Tablets, transfer 1mL of the fil-phase trate to a 200-mL volumetric flask, and dilute with Me-Standard stock solution: 1.16mg/mL of USP dium to volume. Replace the aliquots withdrawn for Ciprofloxacin Hydrochloride RS in Mobile phase analysis with fresh portions of Medium.Standard solution: 0.058mg/mL of USP CiprofloxacinHydrochloride RS in Mobile phase from Standard stocksolution。