A HPTLC DENSITOMETRIC METHOD FOR THE DETERMINATION OF β -SITOSTEROL IN PERGULERIA DAEMIA LEAF AND STEM EXTRACT

HPTLC is a simple, rapid, precise and accurate method with densitometric detection for the determination of beta-sitosterol in a Pergularia daemia Forsk. Petroleum ether and methanol extract of the leaf and stem were powder and used for the experimental work. The separation was performed on silica gel 60 F254 HPTLC plates with Toluene: Ethyl acetate (80:20 v/v), as the mobile phase. The determination was carried out using the densitometric absorbance mode at 690 nm. β-sitosterol response was linear over the range of 100-1000µg/ml, with a correlation coefficient is 0.999. Intraday and interday precision studies showed the relative SD was 0.027. The accuracy of the method was determined, and the average recovery was 99.92%.The limit of quantitation and limit of detection were 8 and 4 mg/ml, respectively. The percentage of beta-sitosterol was found to be 22.16% w/w in unsaponifiable matter of leaf, 33.05% unsaponifiable matter of stem w/w 2.49% w/w petroleum ether extract of leaf 3.28% w/w petroleum ether extract of stem. The method was evaluated for sensitivity, accuracy, precision, and reproducibility. Each analysis by HPTLC is less expensive than current methods. This method is suitable for routine quality control of raw material of the leaves and stem of Pergularia daemia extract and its products.