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S. V. Taralkar and S. Chattopadhyay
Ursolic and betulinic acids are very useful nutraceuticals available in herbs. Their quantitative estimation from the solvent extracts of the herbs is a great challenge. So far all the chromatographic methods available for identification and quantification of these acids are distinctly different, meaning thereby, each acid would require a separate method. Reverse phase (RP-HPLC) method is developed for determination of ursolic acid and betulinic acid from their methanol extract of Vitex negundo Linn leaves. Analysis was carried out using Waters’ symmetry C-18 column with acetonitrile: methanol (80:20) as isocratic elution mode with UV detection (λ=210 nm). The method is pretty linear for ursolic acid in the range of 0.01-0.1 mg/ml (R2 = 0.9961) and for betulinic acid in the range of 0.003-0.018 mg/ml (R2 = 0.999). The peaks of ursolic acid and betulinic acid were confirmed by LC-MS. The method was validated by mixing these acids standards in methanol and found that it is accurate, sensitive and has a good reproducibility.