RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS QUANTITATIVE ESTIMATION OF MEBEVERINE HYDROCHLORIDE AND CHLORDIAZEPOXIDE IN CAPSULES
Keywords:RP-HPLC, Mebeverine, Chlordiazepoxide, Method development, Validation
Objective: To develop an accurate, precise and linear Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method for simultaneous quantitative estimation of Mebeverine hydrochloride and Chlordiazepoxide in MEVA C capsules and validate as per ICH guidelines.
Methods: The optimized method uses a reverse phase column, Enable Make C18G (250 X 4.6 mm; 5Î¼), a mobile phase of triethylammonium phosphate buffer (pH 2.5): acetonitrile in the proportion of 60:40 v/v, flow rate of 1.0 ml/min and a detection wavelength of 240 nm using a UV detector.
Results: The developed method resulted in Mebeverine eluting at 5.8 min and Chlordiazepoxide at 3.9 min. Mebeverine exhibited linearity in the range 250-750Î¼g/ml, while Chlordiazepoxide exhibited linearity in the range 9.25-27.75Î¼g/ml. The precision is exemplified by relative standard deviations of 1.45% for Mebeverine and 1.58% for Chlordiazepoxide. Percentage Mean recoveries were found to be in the range of 98â€102, during accuracy studies. The limit of detection (LOD) for Mebeverine hydrochloride and Chlordiazepoxide were found to be 91.67Âµg/ml and 2.23Âµg/ml respectively, while limit of quantitation (LOQ) for Mebeverine hydrochloride and Chlordiazepoxide were found to be 277.81Âµg/ml and 6.77Âµg/ml respectively.
Conclusion: A simple, accurate, precise, linear and rapid RP-HPLC method was developed for simultaneous quantitative estimation of Mebeverine hydrochloride and Chlordiazepoxide in MEVA C capsules and validated as per ICH guidelines. Hence it can be used for the routine analysis of Mebeverine and Chlordiazepoxide in capusles in various pharmaceutical industries.
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