HPTLC METHOD DEVELOPMENT AND VALIDATION FOR IDENTIFICATION AND QUANTIFICATION OF LUPEOL IN THE LEAVES OF ALSTONIA SCHOLARIS

In ancient literature plant Alstonia scholaris (Apocynaceae) has been considered medicinally important in the treatment of various ailments. Extracts of this plant have shown wide spectra of pharmacological activities like anti-cancer, hepatoprotective, anti-inflammatory, anti-diabetic, etc. These activities can be attributed to the depositories of various phytoconstituents like alkaloids, flavonoids, phenols, triterpenoids, saponins, etc. present in it. Thus, a simple and precise HPTLC method has been developed for identification and quantification of a pentacyclic triterpenoid- lupeol in the leaves of Alstonia scholaris. The developed method was validated following ICH guidelines concerning linearity, range, specificity, precision, accuracy, LOD, and LOQ. After derivatization, the optimized mobile phase Toluene: Ethyl acetate: Formic acid (7:2:1 v/v/v) exhibited good resolution of bands at 580 nm. The R_f value of lupeol was found to be 0.63 and amount quantified in ethanol and chloroform extracts of leaves was 0.31 and 0.35% w/w respectively. The calibration curve of lupeol was linear between 200-1000 ng/band with correlation coefficient 0.9990. LOD and LOQ were 55 and 166.69 ng/band respectively. Recovery was 91.88%. Thus the developed method was found to be specific, precise, and accurate for identification and quantification of lupeol in the leaves of Alstonia scholaris and can be used for routine analysis of the plant.