Determination and quantification of stigmasterol and kaempferol from in vitro and in vivo plant parts of Merremia aegyptia and Merremia dissecta

Abstract

Merremia aegyptia and Merremia dissecta are annual climbing herbs showing high larvicidal and antimicrobial activities. Stigmasterol and Kaempferol were identified and quantified from methanolic extracts of in vivo (leaves, stem, seed) and in vitro (callus) parts using HPTLC densitometric method of both the species. The separation was first performed on TLC aluminium plates precoated with silica gel 60 F . Good separation of stigmasterol was achieved in mobile phase using toluene: methanol (9:1, v/v) at 254 (254, 310nm) and (650nm) and kaempferol using Toluene-acetone-formic acid (4.5: 4.5: 1, v/v) at (254, 366 nm). The instruments CAMAG TLC Scanner LINOMAT- V were used for the present analysis. Determination and quantitation were performed by densitometric scanning at 530 nm for stigmasterol and 254 nm for Kaempferol in reflection/absorbance mode. This method gave compact spots at Rf- 0.92 corresponding to stigmasterol and Rf 0.68-0.71 corresponding to Kaempferol. Highest amount of stigmasterol was calculated from M. aegyptia seed extract (59ng/μl).,Stem extract( 9.8 ng/μl) and callus extract (9.0 ng/μl) concentration and Kaempferol concentration was found highest in M. aegyptia callus (4.9 ng/μl) and M. dissecta seeds (3.6 ng/μl). The present study to screen important bioactive constituents (stigmasterol and kaempferol) can be further utilized for analysis and routine quality control of drugs and formulations prepared from M aegyptia and M. dissecta.