Quantification of β-Sitosterol from Mucuna pruriens by TLC

Mucuna pruriens Linn. one of the popular and important medicinal plants of India is a constituent of more than 200 indigenous drug formulations. β-Sitosterol is one of the most prevalent phytosterols which is ubiquitous throughout the plant kingdom. A sensitive, selective and precise thin-layer chromatographic method has been developed and validated for the analysis of β-sitosterol in Mucuna pruriens roots. Separation and quantification was achieved by TLC using ternary mobile phase of toluene: chloroform; methanol (4:4:1 v/v) (R _F 0.55) on precoated silica gel 60F₂₅₄ aluminium plates and densitometric determination was carried out after derivatization with anisaldehyde-sulphuric acid reagent in reflection/absorption mode at 527 nm. The calibration curve was linear in the concentration range of 100–600 ng spot⁻¹. The method was validated for precision, repeatability and accuracy. The proposed method was found to be simple, precise, specific, sensitive and accurate for the quantification of β-sitosterol.