SIMULTANEOUS DETERMINATION OF QUERCITIN AND AMENTOFLAVONE IN METHANOLIC LEAF EXTRACT OF SEMECARPUS ANACARDIUM (LINN.F.) BY REVERSE PHASE LIQUID CHROMATOGRAPHY Original Article

Objective: To develop a sensitive, simple, rapid and reliable RP-HPLC method that can simultaneously determine Quercitin and Amentoflavone markers in methanolic leaf extract of Semecarpus anacardium (Linn.f.). In this regard HPLC method has been developed and validated as per ICH guidelines. Methods: The reverse phase chromatographic separation was achieved using a Surveyor Agilent 1100 series HPLC system equipped with U.V.detector. The column used for method development and validation was Kromasil 100-5C18 (250x 4.6, 5 μm) as stationary phase with a mobile phase comprising of Water of pH 3(adjusted with O-phosphoric acid): Acetonitrile (55:45 v/v) at a flow rate of 1.0 ml min-1. The injection volume of the sample was 20µl with U.V. detection at 280 nm. The run time of the sample was 10.0 min & the column oven temperature was 250 Results: The proposed RP-HPLC method has been validated as per ICH guidelines for its system suitability, linearity, LOD & LOQ, precision (Intraday & Inter-day), accuracy & robustness. C. Conclusion: The developed RP-HPLC method holds potential for detection and quantification of Quercitin and Amentoflavone markers in methanolic leaf extract of Semecarpus anacardium (Linn.f.).