HPTLC densitometric quantification of stigmasterol and lupeol from Ficus religiosa

This study presents the first report of TLC densitometric method, which has been developed and validated for simultaneous quantification of the two marker compounds (stigmasterol and lupeol) from methanolic extract using the solvent system of toluene:methanol (9:1, v/v). The method employed TLC aluminum plates precoated with silica gel 60 F254 as the stationary phase. Densitometric analysis of stigmasterol and lupeol was carried out in the reflectance mode at 525 nm. The system was found to give compact spots for stigmasterol and lupeol (Rf value of 0.37 and 0.60, respectively). The method was validated using ICH guidelines in terms of precision, repeatability and accuracy. Linearity range for stigmasterol and lupeol was 80–480 ng/spot and 150–900 ng/spot and the contents were found to be 0.06 ± 0.005% w/w and 0.12 ± 0.02% w/w, respectively. The limit of detection (LOD) value for stigmasterol and lupeol were found to be 20 and 50 ng, and limit of quantification (LOQ) value were 60 and 100 ng, respectively. This simple, precise and accurate method gave good resolution from other constituents present in the extract. The method has been successfully applied in the analysis and routine quality control of herbal material and formulations containing Ficus religiosa.