USP 36 Official Monographs / Ciprofloxacin 2995

Result = (rU/rS) × (CS/CU) × 100

Ciprofloxacin Hydrochloride
.

rU = peak area response from the Sample solution

rS = peak area response from the Standard solution
CS = concentration of USP Ciprofloxacin
Hydrochloride RS in the Standard solution
(mg/mL)
CU = concentration of Ciprofloxacin Hydrochloride
in the Sample solution (mg/mL)
Acceptance criteria: 98.0%–102.0% on the anhydrous
C17H18FN3O3 · HCl · H2O 385.82 basis
3-Quinolinecarboxylic acid, 1-cyclopropyl-6-fluoro-1,4- IMPURITIES
dihydro-4-oxo-7-(1-piperazinyl)-, monohydrochloride, Inorganic Impurities
monohydrate; • RESIDUE ON IGNITION 〈281〉: NMT 0.1%
1-Cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl)- • CHLORIDE AND SULFATE, Sulfate 〈221〉: A 375-mg portion
3-quinolinecarboxylic acid, monohydrochloride, monohy- shows no more sulfate than corresponds to 0.15 mL of
drate [86393-32-0]. 0.020 N sulfuric acid (0.04%).
DEFINITION • HEAVY METALS, Method II 〈231〉: NMT 20 ppm
Ciprofloxacin Hydrochloride contains NLT 98.0% and NMT Organic Impurities
102.0% of C17H18FN3O3 · HCl, calculated on the anhy- • PROCEDURE 1: LIMIT OF FLUOROQUINOLONIC ACID
drous basis. Standard solution: Transfer 5.0 mg of USP Fluoroqui-
nolonic Acid RS to a 50-mL volumetric flask containing
IDENTIFICATION 0.05 mL of 6 N ammonium hydroxide, add water to
• A. INFRARED ABSORPTION 〈197K〉 volume, and mix. Transfer 2.0 mL of this solution to a
• B. The retention time of the major peak of the Sample 10.0-mL volumetric flask, and dilute with water to
solution corresponds to that of the Standard solution, as volume.
obtained in the Assay. Sample solution: 10 mg/mL of Ciprofloxacin Hydro-
• C. IDENTIFICATION TESTS—GENERAL, Chloride 〈191〉 chloride in water
Chromatographic system
ASSAY (See Chromatography 〈621〉, Thin-Layer Chromatogra-
• PROCEDURE phy.)
Solution A: 0.025 M phosphoric acid. Adjust with trieth- Mode: TLC
ylamine to a pH of 3.0 ± 0.1. Adsorbent: 0.25-mm layer of silica gel mixture
Mobile phase: Acetonitrile and Solution A (13:87) Application volume: 5 µL
Standard solution: 0.5 mg/mL of USP Ciprofloxacin Hy- Developing solvent system: Methylene chloride,
drochloride RS in Mobile phase methanol, acetonitrile, and ammonium hydroxide
System suitability solution: 0.025 mg/mL of USP (4:4:1:2)
Ciprofloxacin Ethylenediamine Analog RS in Mobile Analysis
phase. Transfer 1.0 mL of this solution to a 10-mL volu- Samples: Standard solution and Sample solution
metric flask, and dilute with Standard solution to Proceed as directed in the chapter. Place a beaker con-
volume. taining 50 mL of ammonium hydroxide in a suitable
Sample solution: 0.5 mg/mL of Ciprofloxacin Hydro- chamber, and then place the plate in the chamber.
chloride in Mobile phase After 15 min, transfer the plate to a suitable chromat-
Chromatographic system ographic chamber, and develop the chromatogram in
(See Chromatography 〈621〉, System Suitability.) the Developing solvent system until the solvent front
Mode: LC has moved about three-fourths of the length of the
Detector: UV 278 nm plate. Remove the plate from the chamber, mark the
Column: 4.6-mm × 25-cm; packing L1 solvent front, and allow the plate to air-dry for about
Column temperature: 30 ± 1° 15 min. Examine the plate under short-wavelength
Flow rate: 1.5 mL/min UV light.
Injection size: 10 µL Acceptance criteria: Any spot from the Sample solution,
System suitability at an RF value corresponding to the principal spot from
Samples: Standard solution and System suitability the Standard solution, is not greater in size or intensity
solution than the principal spot from the Standard solution
[NOTE—The relative retention times for ciprofloxacin (0.2%).
ethylenediamine analog and ciprofloxacin are 0.7 and • PROCEDURE 2
1.0, respectively.] Mobile phase, Standard solution, System suitability
Suitability requirements solution, Sample solution, Chromatographic system,
Resolution: NLT 6 between the ciprofloxacin ethyl- and System suitability: Proceed as directed in the
enediamine analog peak and the ciprofloxacin peak, Assay.
System suitability solution Analysis
Column efficiency: NLT 2500 theoretical plates from Sample: Sample solution
the ciprofloxacin peak, Standard solution Calculate the percentage of each impurity in the por-
Tailing factor: NMT 2.5 for the ciprofloxacin peak, tion of Ciprofloxacin Hydrochloride taken:
Standard solution
Relative standard deviation: NMT 1.5%, Standard Result = (rU/rT) × 100
solution
Analysis rU = peak response of each impurity
Samples: Standard solution and Sample solution rT = sum of the responses of all the peaks
Calculate the percentage of C17H18FN3O3 · HCl in the Acceptance criteria
portion of Ciprofloxacin Hydrochloride taken: Individual impurities: NMT 0.2% for the ciprofloxacin
ethylenediamine analog or any other individual impu-
rity peak
2996 Ciprofloxacin / Official Monographs USP 36

Total impurities: NMT 0.5% Test preparation—Dilute a volume of about 10 µL of Otic

Suspension with Carrier fluid to 25 mL.
SPECIFIC TESTS Procedure—(see Light Obscuration Particle Count Test
• PH 〈791〉: 3.0–4.5, in a 25 mg/mL solution under Particulate Matter in Injections 〈788〉). Analyze the
• WATER DETERMINATION, Method I 〈921〉: 4.7%–6.7% Test preparation using an electronic, liquid-borne particle
ADDITIONAL REQUIREMENTS counting system that employs a light obscuration sensor
• PACKAGING AND STORAGE: Preserve in tight, light-resistant with a suitable sample feeding device. Not less than 99.5%
containers. Store at 25°, excursions permitted between of the particles are ≤25 µm, not less than 99.95% are ≤50
15° and 30°. µm, and not less than 99.995% are ≤100 µm.
• USP REFERENCE STANDARDS 〈11〉 Osmolality 〈785〉: between 270 and 330 mOsmol per kg.
USP Ciprofloxacin Ethylenediamine Analog RS Limit of ciprofloxacin formamide—
1-Cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-[(2-ami- Buffer—Add 6.0 mL of phosphoric acid to 2.0 L of water.
noethyl)amino]-3-quinolinecarboxylic acid Adjust with 50% sodium hydroxide to a pH of 3.0.
hydrochloride.
C15H16FN3O3 · HCl 341.77 Mobile phase—Prepare a filtered and degassed mixture of
USP Ciprofloxacin Hydrochloride RS Buffer and acetonitrile (73:27). Make adjustments if neces-
USP Fluoroquinolonic Acid RS sary (see System Suitability under Chromatography 〈621〉).
Standard solution—Transfer about 25 mg of USP
Ciprofloxacin Formamide RS, accurately weighed, to a
100-mL volumetric flask, and dissolve in and dilute with
methanol to volume. Transfer 3.0 mL of this solution to a
50-mL volumetric flask, and dilute with Mobile phase to vol-
Ciprofloxacin and Dexamethasone Otic
.

ume to obtain a solution having a known concentration of

Suspension about 0.015 mg per mL.
System suitability solution—Transfer about 2.5 mg of USP
» Ciprofloxacin and Dexamethasone Otic Suspen- Dexamethasone RS and about 2.5 mg of USP Ciprofloxacin
sion is a sterile aqueous suspension containing Formamide RS to a 100-mL volumetric flask. Dissolve in
15 mL of methanol, then dilute with Mobile phase to vol-
ciprofloxacin hydrochloride and dexamethasone. ume.
It contains not less than 90.0 percent and not Test solution—Transfer an accurately measured volume of
more than 110.0 percent of the labeled amount freshly mixed Otic Suspension, equivalent to about 6 mg of
of ciprofloxacin (C17H18FN3O3), and not less than ciprofloxacin, to a 10-mL volumetric flask, dilute with Mobile
90.0 percent and not more than 110.0 percent of phase to volume, and mix.
the labeled amount of dexamethasone Chromatographic system (see Chromatography 〈621〉)—The
(C22H29FO5). liquid chromatograph is equipped with a 280-nm detector
and a 3.9-mm × 15-cm column that contains packing L1.
Packaging and storage—Preserve in tight containers, The flow rate is about 1.5 mL per minute. Chromatograph
protected from light. Avoid freezing. the System suitability solution, and record the peak responses
as directed for Procedure: the column efficiency for ciproflox-
USP Reference standards 〈11〉— acin formamide is not less than 2000 theoretical plates; the
USP Ciprofloxacin Ethylenediamine Analog RS resolution, R, between ciprofloxacin formamide and dex-
1-Cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-[(2-ami- amethasone is not less than 8; and the tailing factor for
noethyl)amino]-3-quinolinecarboxylic acid hydrochlo- ciprofloxacin formamide is not more than 2.0. The relative
ride. standard deviation for replicate injections of the Standard
C15H16FN3O3 · HCl 341.77 solution is not more than 2.0%.
USP Ciprofloxacin Formamide RS
USP Ciprofloxacin Hydrochloride RS Procedure—Separately inject equal volumes (about 50 µL)
USP Dexamethasone RS of the Standard solution and the Test solution into the chro-
USP Dexamethasone Acetate RS matograph, record the chromatograms, and measure the re-
sponses for the peaks at the retention time of ciprofloxacin
Identification— formamide. Calculate the percentage of ciprofloxacin forma-
A: The chromatogram of the Assay preparation, obtained mide in the portion of the Otic Suspension taken by the
as directed in the Assay for ciprofloxacin, exhibits a major formula:
peak for ciprofloxacin, the retention time of which corre-
sponds to that obtained in the chromatogram of the Stan- 10(C/VL)(rU / rS)100
dard preparation, obtained as directed in the Assay for
ciprofloxacin. in which C is the concentration, in mg per mL, of USP
B: The chromatogram of the Assay preparation, obtained Ciprofloxacin Formamide RS in the Standard solution; V is
as directed in the Assay for dexamethasone, exhibits a major the volume, in mL, of Otic Suspension taken; L is the la-
peak for dexamethasone, the retention time of which corre- beled amount, in mg per mL, of ciprofloxacin; and rU and rS
sponds to that obtained in the chromatogram of the Stan- are the ciprofloxacin formamide peak responses obtained
dard preparation, obtained as directed in the Assay for dex- from the Test solution and the Standard solution, respectively.
amethasone. Ciprofloxacin formamide is not more than 0.5% of the la-
Sterility 〈71〉—It meets the requirements when tested as beled amount of ciprofloxacin.
directed for Membrane Filtration under Test for Sterility of the Ciprofloxacin related compounds—
Product to be Examined. Procedure—From the chromatogram of the Assay prepara-
pH 〈791〉: between 3.8 and 4.8. tion, obtained as directed in the Assay for ciprofloxacin,
Particle size— measure the responses for the ciprofloxacin ethylenediamine
analog and the other minor peaks. Calculate the percentage
Carrier fluid—Heat Purified Water to a temperature of 40°
to 50°, add 100 mg of dexamethasone per L while stirring,
cool to room temperature while stirring, pass through a 0.2-
µm filter, and store in a clean, covered container.