Simultaneous Estimation of Epicatechin, Trigonelline, Stigmasterol, and Gymnemagenin in Polyherbal Products: A Bioanalytical RP-HPLC Approach

Abstract

The present study is about the development of bioanalytical method for the simultaneous estimation of Epicatechin (EPI), Trigonelline (TRG), Stigmasterol (STIG) and Gymnemagenin (GYM) in the polyherbal formulation using HPLC.  

The polyherbal formulation were prepared by extracting Pterocarpus marsupium, Gymnema sylvestre, Trigonella foenumgrecum, Momordica charantia herbs which has wide acceptance in the treatment of diabetes and other chronic disorders. To investigate the pharmacokinetic of this developed polyherbal formulation, current work have devised a simple reverse phase high-performance liquid chromatography (RP-HPLC) method for the estimation of EPI, TRG, STIG and GYM in rat plasma. In doing this the plasma samples were spiked with the marker compounds and then extracted by solid phase extraction with Phenomenex Strata-X 33µm cartridges. Further, chromatographic separation was performed on C18 column using isocratic acetonitrile:water composition.  The developed method was validated for accuracy, precision, linearity and recovery. Linearity studies were found to be acceptable over the range of 0.1–6 lg/ml. The method was successfully applied for the analysis of rat plasma sample for the application in pharmacokinetic study, drug interaction, bioavailability and bioequivalence. The mean %  recovery from plasma  of  EPI were 92.8, 91.2 & 94.7, for TRG 91.5, 92.8 & 93.4, STIG 91.8, 92.5 & 91.8 and GYM 92.8, 93.1 & 92.5 % for LQC, MQC & HQC respectively.  The recovery of EPI, TRG, STIG and GYM were found to be within the limit of acceptance.