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Abstract

SIMULTANEOUS QUANTIFICATION OF LUPEOL, Β-SITOSTEROL AND OLEANOLIC ACID USING VALIDATED HPTLC METHOD FROM NYCTANTHES ARBOR-TRISTIS AND ITS MARKETED FORMULATION

Vikas V. Vaidya*, Pushkar M. Pradhan and Prakash Kondalkar

ABSTRACT

A simple, accurate and reproducible HPTLC densitometric method has been developed for the simultaneous quantification of Lupeol, β- sitosterol and Oleanolic acid from the methanolic extract of leaves of plant Nyctanthes arbor-tristis and its marketed formulation. The mobile phase n-Hexane: Ethylacetate: Formic acid (8: 2: 0.5 v/v/v) gave good separation and resolution for the mentioned compounds. The CAMAG HPTLC system was used for the analysis. The separated compounds were visualized at 540 nm after derivatization using 10% Methanolic sulphuric acid reagent. Rf value of Oleanolic acid, β- sitosterol and Lupeol was found to be 0.40, 0.49 and 0.574 respectively. The quantity of Lupeol was found to be 0.03124% and 0.008128%, β-sitosterol was 0.0777% and 0.003054% and Oleanolic acid was 0.1062% and 0.008369% in plant and formulation respectively. The developed method was then validated in accordance with the ICH guidelines in terms of specificity, linearity, LOD, LOQ, precision and recovery. The validated method was successfully applied for quantification of three components in a formulation containing Nyctanthes arbor-tristis extract. This method can also be used as a quality control tool for other formulations or dietary supplements containing the extract of Nyctanthes arbor-tristis.

Keywords: Nyctanthes arbor-tristis, Lupeol, ?-sitosterol, Oleanolic acid, simultaneous quantification.


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