Abstract
A simple normal-phase high-performance thin-layer chromatography (HPTLC) method has been developed for the quantification of β-sitosterol in hairy root cultures of Clitoria ternatea vis-a-vis natural plant parts. Chromatographic separation was achieved on normal-phase TLC plates by using optimized mobile phase of n-hexane. acetone (8:2, v/v). Densitometric scanning was performed after postchromatographic derivatization of the plate at 414 nm. The limits of detection and quantification were found to be 40 ng and 100 ng spot−1, respectively. A linear response of calibration line was observed over the range of 100 to 500 ng spot−1 with a correlation coefficient of r2 = 0.999. Recovery values from 97.22 to 98.51% with small coefficient of variation showed excellent accuracy of the method. The method was validated according to the International Conference on Harmonization (ICH) protocol. The method enables excellent separation and accurate quantification of β-sitosterol in different plant parts of the two flor...
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