Analytical Stability Indicative Method Development and Validation by High Pressure Liquid Chromatography for Assay in Ciprofloxacin hydrochloride Drug Substances

Abstract

Ciprofloxacin is an antibiotic generic drug that can treat a number of bacterial infections. It is a second generic fluoroquinolone. This work presents a simple, sensitive and stability indicative method for assay determination of drug substances ciprofloxacin hydrochloride by high pressure liquid chromatography (HPLC). Separation of impurities from the main drug substances and accurate assay quantification with a simple method is difficult by Spectrophotometry. The separation of co-eluents and impurities from the ciprofloxacin drug substances was archived by HPLC with simple mobile phase 0.15% orthophosphoric acid adjusted pH to 3.0 with triethylamine and acetonitrile using a gradient program, column inertsil C18, 250 mm × 4.0 mm, 5 μm. Flow rate was 0.7 mL/min, the column temperature and injection volume were 35°C and 10.0 μL. Chromato-graphic analysis was carried out at wavelength 278 nm. The developed method was validated according to the international conference on harmonization (ICH) guidelines regarding: Precision, specificity by degradation, linearity, accuracy, range and robustness. The proposed method showed good linearity (correlation coefficient and regression coefficient were not less than 0.999 and 0.998) in the range of 50% to 150% of working concentration. The recovery at 50%, 100% and 150% of working concentration level was within 98.0% to 102.0%. The range of the method is concluded that developed method is from 50% to 150% of target concentration for ciprofloxacin hydrochloride. The applicability of the proposed method was verified through the analysis of the samples and percentage recoveries in the range of 98% to 102% were obtained without any interference detected at the main peak in the chromatogram. The validated method is highly selective, simple, accurate, cost effective, and it is applicable for stability studies and routine quality-control analysis in the pharmaceutical industries.