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Department of Pharmaceutical Quality Assurance, Aditya Bangalore Institute of Pharmacy Education & Research.
The present study aimed to develop and validate a simple, precise, accurate, robust, and economical reverse-phase high-performance liquid chromatographic (RP-HPLC) method for the quantitative estimation of Ciprofloxacin in tablet dosage forms. Chromatographic separation was achieved using a C18 column (250 mm × 4.6 mm, 5 µm). The mobile phase consisted of acetonitrile and phosphate buffer (pH 3.5) in the ratio of 60:40 (v/v), delivered at a flow rate of 1.0 mL/min. Detection was performed at 278 nm with an injection volume of 20 µL. The method exhibited excellent linearity over the concentration range of 10–100 µg/mL with a correlation coefficient (R²) of 0.999. The mean recovery was found to be 99.1%, indicating good accuracy. Intra-day and inter-day precision studies showed %RSD values of 0.82% and 0.95%, respectively. The LOD and LOQ values were 0.25 µg/mL and 0.80 µg/mL, respectively. System suitability parameters showed a retention time of approximately 4.2 min and a tailing factor of 1.2.
Pharmaceutical analysis plays an essential role in ensuring the quality, safety, and efficacy of pharmaceutical products. Among various analytical techniques, High-Performance Liquid Chromatography (HPLC) is one of the most reliable and widely accepted methods for the
qualitative and quantitative analysis of pharmaceutical compounds.
Ciprofloxacin is a second-generation fluoroquinolone antibiotic extensively used for the treatment of bacterial infections caused by Gram-positive and Gram-negative microorganisms. It acts by inhibiting bacterial DNA gyrase and topoisomerase IV, thereby preventing DNA replication and resulting in bacterial cell death.
Due to the widespread clinical use of Ciprofloxacin, reliable analytical methods are required for quality control and regulatory compliance. Therefore, the present investigation focused on the development and validation of an RP-HPLC method for the estimation of Ciprofloxacin in tablet dosage forms according to ICH guidelines.
MATERIALS AND METHODS
Chemicals and Reagents
Instrumentation
Chromatographic Conditions
|
Parameter |
Condition |
|
Column |
C18 (250 mm × 4.6 mm, 5 µm) |
|
Mobile Phase |
Acetonitrile : Phosphate Buffer (60:40 v/v) |
|
pH |
3.5 |
|
Flow Rate |
1.0 mL/min |
|
Detection Wavelength |
278 nm |
|
Injection Volume |
20 µL |
|
Run Time |
10 min |
|
Column Temperature |
Ambient |
Preparation of Standard Solution
Accurately weighed 100 mg of Ciprofloxacin reference standard was dissolved in mobile phase and diluted to 100 mL to obtain a stock solution of 1000 µg/mL.
Preparation of Sample Solution
Ten tablets were weighed and powdered. A quantity equivalent to 100 mg of Ciprofloxacin was dissolved in mobile phase, sonicated for 15 minutes, filtered through a 0.45 µm membrane filter, and diluted appropriately.
Method Validation
Linearity
Calibration standards ranging from 10–100 µg/mL were prepared and injected into the HPLC system.
|
Concentration (µg/mL) |
Peak Area |
|
10 |
12545 |
|
20 |
25110 |
|
40 |
50230 |
|
60 |
75340 |
|
80 |
100520 |
|
100 |
125600 |
Correlation coefficient (R²): 0.999
Accuracy
Recovery studies were performed at three levels.
|
Level |
% Recovery |
|
80% |
98.6 |
|
100% |
99.2 |
|
120% |
100.3 |
Mean recovery: 99.1%
Precision
|
Parameter |
%RSD |
|
Intra-day Precision |
0.82 |
|
Inter-day Precision |
0.95 |
Sensitivity
|
Parameter |
Value (µg/mL) |
|
LOD |
0.25 |
|
LOQ |
0.80 |
Robustness
Small deliberate changes in flow rate, mobile phase composition, and wavelength showed no significant effect on chromatographic performance.
System Suitability
|
Parameter |
Result |
|
Retention Time |
4.2 min |
|
Theoretical Plates |
>3000 |
|
Tailing Factor |
1.2 |
|
Resolution |
>2 |
RESULTS AND DISCUSSION
The developed RP-HPLC method successfully produced a sharp and symmetrical peak of Ciprofloxacin with acceptable retention time and excellent peak resolution. The method demonstrated excellent linearity (R² = 0.999), indicating a direct proportional relationship between concentration and detector response.
Recovery studies confirmed the accuracy of the method, with percentage recoveries within acceptable limits. Precision studies exhibited %RSD values below 2%, demonstrating good reproducibility. The low LOD and LOQ values indicated adequate sensitivity of the method.
Robustness studies showed that minor changes in chromatographic conditions did not significantly affect analytical performance. Assay of marketed tablet formulations indicated drug content within pharmacopeial limits, confirming the suitability of the method for routine quality control applications.
CONCLUSION
A simple, rapid, precise, accurate, and robust RP-HPLC method was successfully developed and validated for the estimation of Ciprofloxacin in tablet dosage forms. The method complied with ICH validation requirements and demonstrated excellent analytical performance. The validated method can be effectively employed for routine quality control analysis, stability studies, and pharmaceutical research involving Ciprofloxacin formulations.
REFERENCES
Manmohan Singh, Vipin Kumar Singhal, Narendra Sharma, Vishal Garg, Development and Validation of a Reverse Phase High-Performance Liquid Chromatographic Method for Estimation of Ciprofloxacin in Tablet Dosage Form, Int. J. of Pharm. Sci., 2026, Vol 4, Issue 7, 2403-2406, https://doi.org/10.5281/zenodo.21323476
10.5281/zenodo.21323476