High-performance thin-layer chromatography densitometric determination of beta-sitosterol in Phyllanthus species

An HPTLC densitometric method was established for the determination of beta-sitosterol in in vitro cultures of some species of the genus Phyllanthus. Two derivatization reagents commonly used in TLC for the visualization and detection of sterols, namely, anisaldehyde reagent and 5% phosphomolybdic acid, were compared with vanillin reagent. The densitometric quantification of beta-sitosterol was carried out on HPTLC Si60 F254 plates with the mobile phase chloroform-hexane-methanol (65 + 30 + 5, vlv/v) at 525 nm. The method was validated for each derivatization reagent in terms of linearity, precision, repeatability, intra- and interday precision, LOD, and LOQ. The presence of beta-sitosterol was revealed in all analyzed plant material. The concentrations of beta-sitosterol determined ranged from 0.48 to 2.75 mg/g (dry weight). In addition, traces of beta-amyrin were detected in some plant samples.