High-Performance Thin-Layer Chromatography Method Development for Estimation of Triterpenoids in Calotropis Gigantea Roots

Abstract

Development of analytical method on high performance thin layer chromatography for estimation of content of marker compound, preferably bioactive constituent, in traditional herb has become a rational approach for its standardization. The literature does not reveal any report on phytochemical investigation to standardize Calotropis gigantea roots on the basis of marker compounds. Non-availability of such study on Calotropis gigantea prompted us to develop high performance thin layer chromatography method for the estimation of triterpenoids (taraxerol and lupeol), which are bioactive compounds of the plant. Numerous solvent systems were tried to resolve these compounds in methanol extract of Calotropis gigantea roots but best resolution of taraxerol and lupeol was obtained using toluene:ethyl acetate:methanol (8:1:1) and toluene:chloroform (3:7), respectively, as solvent systems. The developed thin layer chromatography plates were derivatized by treating with anisaldehyde-sulphuric acid reagent followed by heating. The visible coloured bands of triterpenoids were scanned at 550 nm. The percentage content of taraxerol and lupeol was 0.0086 % and 0.0093 % w/w, respectively, in Calotropis gigantea roots. The consistent and reproducible therapeutic effects of herbal preparations containing Calotropis gigantea roots can be achieved if marker based standardized raw material of the plant is used.