"Development and Validation of HPTLC Method for the Determination of Clopidogrel bisulphate in Pharmaceutical Dosage Form and Simulated Biofluids”

Abstract

A simple, rapid, and sensitive high performance thin layer chromatographic method has been developed for the quantification of clopidogrel in simulated biofluids and pharmaceutical dosage form and validated according to standard guidelines. The separation of clopidogrel bisulphate was achieved using aluminium-backed layer of silica gel 60 F254. The toluene: acetonitrile: methanol: acetone (5.5:3:1:0.5 v/v/v/v) as the mobile phase used. Densitometric analysis was carried out in the absorbance mode at 230 nm. The system was found to give compact spots for clopidogrel bisulphate (Rf value of 0.81). Sample preparation of drug from simulated biofluids was carried out with 0.45 µm nylon syringe filter. The limit of detection was 7.36 ng/band and limit of quantification was 22.89 ng/band. The linear regression analysis data for the calibration plots showed good linear relationship with r 2 = 0.9996 in the concentration range 25-250 ng per spot for both standard solutions and spiked simulated biofluids. The intra-day and inter-days precision (RSDs) was less than 2 % and the accuracy was more than 99%. The method could be applied for the estimation of clopidogrel bisulphate in simulated biofluids as well as to pharmaceutical dosage forms.
Abbreviations – Clopidogrel bisulphate (CLB), Simulated biofluids (SBFs)