Development and Validation of HPTLC Method for Simultaneous Estimation of Berberine, Gallic Acid and Ursolic Acid in a Polyherbal Blend

Abstract

A sensitive high-performance thin-layer chromatography method was developed for simultaneous estimation of berberine, gallic acid and ursolic acid in a polyherbal blend and validated as per ICH guidelines. Polyherbal blend was prepared using widely recommended herbal plants for platelet augmentation activity viz. Carica papaya, Berberis aristata, Ocimum Sanctum, and Tinospora Cordifolia. The optimized separation was obtained with TLC aluminum plates pre-coated with silica gel G60 F254 as a stationary phase and a solvent system containing Toluene : Ethyl acetate : Methanol : Formic acid (3:3:0.2:0.1 v/v/v/v). Berberine and gallic acid were found to demonstrate linearity in the range of 1μg/band – 6μg/band and ursolic acid in the range of 20 μg/band – 100 μg/band with the regression coefficient in acceptable limits. The method was also found to be specific and precise one. The accuracy of the developed method at 80 %, 100 % and 120 % levels was found to be within limits and % RSD was found to be less than 2. LOD and LOQ of all three standards were also determined. Blend was also quantified for the amount of berberine, gallic acid and ursolic acid presence and was found to be 37.8 μg, 46.1 μg and 108 μg per mg, respectively.