Identification and Qualitative Analysis of β-Sitosterol and Some Phytoestrogens in In Vivo and In-Vitro Samples of Lepidium sativum: A Semi-Arid Bone Healing Plant

Abstract

Dry powder of different plant parts of Lepidium sativum (Garden Cress) are known to be used for treatment of fracture (bone healing) from ancient times. Seeds as in vivo plant part and callus as in vitro plant parts were chromatographically tested to identify and estimate β-sitosterol and various phytoestrogens. Plant-derived sterols and estrogens in tissues and oilseeds of Lepidium sativum were isolated by solvent extraction method. β-sitosterol, diadzein and formononetin were estimated qualitatively and quantitatively through TLC and HPTLC techniques. In vitro grown callus was compared biochemically with in vivo plant parts. CAMAG HPTLC was used and the developed plates were photo-documented using UV and white light. The amount of β-sitosterol was estimated to be about 0.20% w/w for seed powder and 0.024% w/w for callus powder. Daidzein and formononetin isolated from the samples of Lepidium sativum showed Rf values 0.73 and 0.67 and peak areas were 546.8 and 1033.5, respectively. Chloroform: methanol (8:2) gave excellent results in the present investigation for all samples. Results indicated that applied assay is accurate and reliable for the determination of phytoestrogens in the plant.