Abstract

QUANTIFICATION OF LUPEOL AND STIGMASTEROL FROM METHANOLIC EXTRACT OF SEMECARPUS ANCARDIUM LINN.F BY HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY

Mohmad Vasim Sheikha,*, Navin Devadigab and Dr. Manish Hatec

ABSTRACT

Semecarpus ancardium Linn.F belongs to family Anacardeciae is well known for its use in ayurveda, siddha and unani as a potent medicinal herb since ancient times. According to literature survey HPTLC method was developed on the quantitation of Lupeol and Stigmasterol but simultaneous work was not done the Semecarpus ancardium Linn.F therefore a need was felt to develop a precise, simple and accurate method for analysis of these phytoconstituents. In the present research work, normal mode of separation was used, where stationary phase was comparatively more polar than mobile phase. A quantitative determination of Lupeol and Stigmasterol was done by HPTLC using Toluene, Methanol and Formic acid as a mobile phase in the ratio of (6.5 cm3: 3.0cm3: 0.5cm3) v/v/v. Ascending mode of chromatographic development was chosen for present work. The plates were developed in a CAMAG glass twin trough chamber (20 x 10 cm and 10 x 10 cm) lined with filter paper and was previously saturated with mobile phase vapours for 20 minutes. The development was carried up to a distance of 60 mm .They were then dried with ambient air for 10 minutes. The plate was then developed and scanned under optimised chromatographic conditions. The Lupeol and Stigmasterol showed maximum absorbance at 530 nm. Therefore, 530 nm was selected for the detection and quantification of Lupeol and Stigmasterol from the aerial powder of Semecarpus ancardium Linn.F. The plates were scanned using CAMAG TLC Scanner 3, with win CATS software, version 1.4.4, in absorbance/reflectance mode, using tungsten lamp at the wavelength of maximum absorption of Lupeol and Stigmasterol at λ = 530nm, slit size 6.0 x 0.45 mm. The Rf values of Lupeol and Stigmasterol were found to be 0.66 and 0.53 respectively. Linearity of r2 = 0.998 and 1.0, LOD 0.1 and 0.05μg/band, LOQ 0.2 and 1.0μg/band and recovery of 99.59% and 104.79% respectively were satisfactory for Lupeol and Stigmasterol. The HPTLC method was validated as per the ICH guideline. The developed simple, precise, specific, sensitive and accurate method can be used for routine quality control of raw material of Semecarpus ancardium Linn.F as well as formulation containing any of these bioactive compounds.

Keywords: HPTLC, Semecarpus ancardium Linn. F, Lupeol and Stigmasterol.