Bioanalytical Methods for Estimation of Gliclazide in Human Plasma

Abstract

The objective of this study is to estimate gliclazide in human plasma by both HPLC and HPTLC method. For HPLC method, SunQ C18 column was used as stationary phase and Methanol: 0.1% Formic acid in Water was used as mobile phase in ratio of 70:30 v/v. For HPTLC method, TLC plate precoated with silica gel 60 F254was used as stationary phase and mobile phase selected was Toluene: Methanol: Ethyl acetate in ratio of 7:2:1 v/v/v. Simple protein precipitation method was employed as sample pretreatment in both HPLC and HPTLC methods. The detection wavelength set was 228 nm. Molnupiravir and Glipizide were used as internal standard for HPLC and HPTLC method respectively. With optimised HPLC method, Gliclazide eluted at 7.5±0.2 mins and the Rf value found was 0.49±0.06 for HPTLC. Both the methods were found to be linear over the range of 2-10 µg/ml. The r 2 value found for HPLC and HPTLC method was 0.9842 and 0.9835 respectively. Both of the methods were validated for as per USFDA guidelines for bioanalytical method. The developed methods were found to be simple, accurate, precise and economical for estimation of Gliclazide in human plasma.