DEVELOPMENT AND VALIDATION OF ANALYTICAL METHOD FOR SIMULTANEOUS ESTIMATION OF ELLAGIC ACID AND BERBERINE IN POLYHERBAL FORMULATION BY RP-HPLC

Abstract

An easy, selective, rapid, specific and low cost reverse phase HPLC technique has been developed for the simultaneous estimation of ellagic acid and berberine from a polyherbal formulation. The method was carried out on a Hemochrom Intsil [C18 (250 X 4.6 mm, 5 µ)] column with the mobile phase consisting of methanol: 0.1 % orthophosphoric acid (80:20V/V) at a flow rate of 0.8 mL/min. Detection was carried out at 234 nm. The retention times of ellagic acid and berberine were 4.00 and 2.97 min, respectively. The regression analysis data indicated a good linear relationship for the calibration plots for ellagic acid and berberine in the range of 2-8 µg/mL and 20-80 µg/mL and the regression coefficient was 0.997 and 0.9966, respectively. The accuracy and reliability of the developed method were established by means of comparing numerous validation parameters like linearity, precision, accuracy, robustness, specificity, the limit of quantification and limit of detection, in keeping with the ICH guidelines. The proposed method can be used for the estimation of these markers in other polyherbal formulations.