Development of validated HPTLC method for quantification of stigmasterol from leaf and stem of Bryophyllum pinnatum

Abstract This study presents the report of TLC densitometric method, which has been developed and validated for quantification of stigmasterol from petroleum ether extract of leaves and stems of Bryophyllum pinnatum . The separation was performed on TLC aluminum plates precoated with silica gel 60 F 254. Good separation was achieved in mobile phase using Chloroform:Ethanol (9.8:0.2 v/v). Determination and quantitation were performed by densitometric scanning at 490 nm in reflection/absorbance mode. This method gave compact spots at R f 0.45 corresponding to stigmasterol. The method was validated using ICH guidelines in terms of precision, repeatability and accuracy. Linearity range for stigmasterol was 50–300 ng/spot with correlation coefficient R 2  ± SD = 0.990 ± 5.05% in the concentration range 50–300 ng/spot. The LOD and LOQ were found to be 15 and 45 ng, respectively. The contents in crude extract obtained from leaf and stem were found to be 0.1545 ± 0.005% w/w and 0.1383 ± 0.002% w/w respectively. HPTLC method was found to be simple, precise, accurate and convenient method for rapid screening of active constituents present in petroleum ether extracts of plant and can be used for analysis and routine quality control of herbal material and formulations containing B. pinnatum .