ISOLATION, IDENTIFICATION, AND SIMULTANEOUS QUANTIFICATION OF FIVE MAJOR FLAVONOIDS IN EPIMEDIUM ELATUM BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

A simple and reliable HPLC-UV-DAD method was developed for rapid resolution, identification, and quantitation of prenylated flavonoids in Epimedium elatum, a plant endemic to Kashmir Himalayas. Five major prenylated flavonoids namely epimedoside A, epimedin A, epimedin B, epimedin C, and icariin were isolated using repeated column chromatography and simultaneously quantified using HPLC in both underground and aerial parts of E. elatum. All calibration curves showed good linearity (r(2)>0.998) within test ranges and recoveries were 96.8 to 101.2%. The optimized method was successfully applied for the analysis of five major flavonoids in 18 samples of E. elatum collected from three different ecogeographical zones in three different harvesting seasons. The contents of five investigated compounds were greatly variant ranging from 8.73% to 10.96% in aerial parts and 2.52% to 5.06% in underground parts. The concentration of epimedin C was found to be the highest (6.42%) among the five quantified compounds in one of the accessions. The method established in this paper is simple and reliable and could be used for the quality control of E. elatum.