Simultaneous Estimation of Stigmasterol and Lupeol in Adenanthera pavonina using HPTLC Method

Abstract

Adenanthera pavonina , a well-known medicinal plant has been used traditionally to treat diarrhea and inflammation. Its medicinal properties result from its active phyto-constituents, including non-protein amino acids, sterols, dulcitol, and fixed oil. Stigmasterol (STIGMA) and lupeol (LPL) are significant phytosterols in the plant’s bark as they possess anti-inflammatory activities that may be used in multiple therapies. To design an accurate HPTLC method for the simultaneous estimation of lupeol and stigmasterol from the bark of Adenanthera pavonina. The STIGMA and LPL were isolated using silica gel (60–120 #) column chromatography and estimated using HPTLC. The TLC aluminium plates were pre-coated with silica gel 60F254; the mobile phase comprised toluene and methanol in the ratio of 92:8v/v, and the compounds were detected using the vanillin-phosphoric acid reagent by densitometric scanning at 550 nm. The R f value for STIGMA and LPL was 0.430.02and0.550.02 with linearity in the range of 100– 500 ng/spot and 400-2000 ng/spot, respectively. The mean % recovery was in the range of 93.8–97.2 % for STIGMA and 94.9 –95.6 % for LPL with LOD and LOQ values of 11.74 and 35.58 ng/spot for STIGMA and 90.44 and 274.06 ng/spot for LPL, respectively. Our HPTLC method was accurate and precise for simultaneous estimation of STIGMA and LPL from the bark of Adenanthera pavonina, thereby making it effective in quality control investigations of the herbal formulations. Keywords: Adenantherapavonina; HPTLC; Stigmasterol; Lupeol