SIMULTANEOUS QUANTIFICATION OF DIOSGENIN, OLEIC ACID, HARMINE AND KAEMPFEROL BY RP-HPLC IN THE FRUIT EXTRACT OF TRIBULUS TERRESTRIS L. AND ITS FORMULATION

A simple, accurate and reproducible RP-HPLC method has been developed for the simultaneous quantification of Diosgenin, Oleic acid, Harmine and Kaempferol from the methanolic extract of fruits of plant Tribulus terrestris L. and its marketed formulation. The Agilent 1290 Infinity II HPLC system was used for the analysis. The optimized conditions (Mobile phase A: 0.01 M Di-Potassium hydrogen phosphate buffer; pH 7.0 adjusted with orthophosphoric acid And Mobile phase B Acetonitrile: Methanol (80:20), gradient elution system on a C8 reversedphase column with a flow rate of 1 mL/min. The separated compounds were detected at 201 nm using Agilent 1290 DAD FS ( Diode Array Detector) The quantity of Harmine was found to be 0.013 % and 0.006%, Kaempferol was 0.011% and 0.005%, Oleic acid was 0.261 % and 0.115 % and Diosgenin was 0.084 % and 0.019% in plant and formulation respectively. The developed method was then validated and applied for quantification of four components in a formulation containing Tribulus terrestris L. extract. This method can also be used as a quality control tool for other formulations or dietary supplements containing the extract of Tribulus terrestris. Keywords: Tribulus terrestris L., Harmine, Kaempferol, Oleic acid, Diosgenin, Simultaneous quantification