Abstract

HPTLC FINGERPRINTING AND QUANTIFICATION OF Β SITOSTEROL IN EXTRACT AND POLYHERBAL FORMULATIONS

Prof. Mayur R. Bhurat*, Yogesh M. Bagad, Dr. Surajj Sarode and Arvind R. Umarkar

Abstract

A new, simple, sensitive, precise and robust high-performance thin layer chromatographic (HPTLC) method was developed for the estimation of β sitosterol in herbal extracts and polyherbal formulation. Analysis of β sitosterol was performed on TLC aluminium pre-coated plates with silica gel 60F-254 as stationary phase. Linear ascending development was carried out in twin trough glass chamber saturated with mobile phase consisting of Toluene: ethyl acetate: formic acid (5:2:1 v/v/v) for β sitosterol at room temperature (25±2oC). After development the plate was derivatized with 20% ethanolic H2SO4. Camag TLC scanner III was used for spectrodensitometric scanning and analysis of plate in absorbance mode at 364 nm for β sitosterol. The system was found to give compact spots for β sitosterol (Rf 0.42). The data for calibration plots showed good linear relationship with r2 = 0.9984 in the concentration range of 60–360 ng for β sitosterol. According to international conference on harmonization (ICH) guidelines the present method was validated for precision, repeatability and recovery. The limits of detection and quantification were determined. Statistical analysis of the data showed that the method is reproducible and selective for estimation of β sitosterol.

Keywords: High-performance thin layer chromatography; Method validation; ? sitosterol; Polyherbal formulations.