Journal of chromatographic science

Simultaneous HPTLC analysis of ursolic acid, betulinic acid, stigmasterol and lupeol for the identification of four medicinal plants commonly available in the Indian market as Shankhpushpi.

PMID 25217706


In this study, we investigated a new, simple, sensitive, selective and precise high-performance thin-layer chromatography (HPTLC) fingerprint and quantitative estimation method for the analysis of ursolic acid, betulinic acid, stigmasterol and lupeol in Shankhpushpi botanicals. Linear ascending development was carried out in a twin trough glass chamber saturated with petroleum ether-ethyl acetate-toluene (7:2:1, v/v/v). The plate was dried, sprayed with anisaldehyde reagent and analyzed by CAMAG TLC scanner III at 580 nm. The system was found to give compact spots for ursolic acid (0.21), betulinic acid (0.29), stigmasterol (0.33) and lupeol (0.50). The relationship between the concentration of standard solutions and the peak response is linear within the concentration range of 100-600 ng/spot for ursolic acid, betulinic acid, stigmasterol and lupeol. The concentration of 134.2 and 146.1 mg of ursolic acid per gram of Clitorea ternatea (CT) and Canscora decussata (CD); 110.6 mg of betulinic acid per gram of EA; 92.75, 154.95, 31.947 and 39.21 mg of stigmasterol per gram of Evolvulus alsinoides (EA), Convolvulus pluricaulis (CP), CT and CD; 30.12 mg of lupeol per gram of CT were found. The proposed HPTLC method may use for routine quality testing and identification of Shankhpushpi botanicals.