DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR THE ESTIMATION OF GLICLAZIDE

The aim of the present work was to develop and validate a method for the estimation of gliclazide by reverse phase HPLC method. A validation protocol was followed to develop an analytical method, using Grace Vydac C18 column (250 x 4.6 mm, 5m) as stationary phase and acetonitrile:ammonium acetate buffer as mobile phase. The mobile phase pH, ratio of mobile phase, ionic strength of buffer, the flow rate was optimized and the method was validated for its linearity, accuracy, precision, robustness. The retention time was within 15 min, acetonitrile:ammonium acetate buffer (50:50) with an ionic strength of 20 mM and a flow rate 1ml/ml was selected for elution. The method was linear in the range from 10–70 μg/ml with (r2) 0.9995, accuracy was 99.62±1.43 (% Mean ±SD), robustness was found to be 0.92%, repeatability and intermediate precision was found to be 0.37% and 0.58% respectively, LOD and LOQ was found to be 0.1 and 0.5 μg /ml respectively. The developed method was found to be linear, accurate, robust and precise and the values obtained were within acceptable values.