Thin-layer chromatographic separation and a validated high-performance thin-layer chromatography method for the quantification of stigmasterol in 48 different extracts of Lasiurus scindicus Henrard (Poaceae)

Abstract

The present study is the first attempt on the phytochemical analysis of Lasiurus scindicus Henrard, a desert-specific plant, through high-performance thin-layer chromatography (HPTLC). This technique has been developed and validated for the quantification of stigmasterol from 48 extracts, i.e., four different plant parts with six different solvents (butanol, chloroform, ethanol, hexane, methanol, and petroleum ether) in both hot and cold macerations. This method gave the best resolution bands with toluene‒methanol‒formic acid (9:1:0.3, v/v) for 25 min as the mobile phase at retention factor (RF) of 0.27 for stigmasterol. The method was validated using the International Conference on Harmonization (ICH) guidelines in terms of precision, repeatability, and accuracy. The linearity range for stigmasterol was found to be 2–12 μg spot−1. Both hot and cold extractions with six different solvents were compared. The highest concentration of stigmasterol was found in the leaves of L. scindicus in cold hexane extrac...