DEVELOPMENT OF ANALYTICAL METHOD AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF CLOBETASOL PROPIONATE AND KETOCONAZOLE IN PHARMACEUTICAL CREAM FORMULATION BY RP-HPLC METHOD

A simple, accurate rapid and precise has been developed and validated for simultaneous estimation of Clobetasol Propionate and Ketoconazole in Pharmaceutical Cream Formulation by RP-HPLC Method. The successful estimation was carried out of the drug product is developed on the ODS Qualisil Column C18 (250 mm x 4.6 mm, 5μm or equivalent) at ambient temperature using Methanol: Acetonitrile: Phosphate buffer (50:20:30 v/v/v) as mobile phase composition. The flow rate was adjusted to 1.5mL/minute and the absorption maxima were observed on UV detector at 254 nm. Retention Time for Clobetasole Propionate 11.1±0.1min and Ketoconazole 12.2±0.1min. The linearity was obtained in the concentration range of 6-14µg/mL and 120-280µg/mL for Clobetasole Propionate and Ketoconazole respectively. Mean percentage recoveries were 99.16% for Clobetasole Propionate and 99.17% for Ketoconazole. The LOD of Clobetasole Propionate and Ketoconazole were found to be 0.7μg/mL and 2.0μg/mL whereas the LOQ was 3.0μg/mL and 10.0μg/mL respectively. Percentage relative standard deviation of percent assay values for replicate sample preparation was 0.28% for Clobetasole Propionate and 0.47% for Ketoconazole. The method was robust with respect to change in flow rate, temperature and composition of mobile phase. The method was validated statistically and applied successfully for the determination of Clobetasole Propionate and Ketoconazole. Validation studies revealed that method is specific, rapid, reliable, and reproducible. The high recovery and low relative standard deviation confirm the suitability of the method for the routine determination of Clobetasole Propionate and Ketoconazole in pharmaceutical cream formulation. Keywords: Clobetasol Propionate, Ketoconazole, RP – HPLC, Method Validation