Separation and purification of epimedin A, B, C, and icariin from the medicinal herb Epimedium brevicornum maxim by dual-mode HSCCC.

Epimedium brevicornum Maxim is a famous medicinal herb which has been widely used for the treatment of impotence, infertility, osteoporosis, cardiovascular diseases, amnesia, and senile functional diseases. A dual-mode high-speed counter-current chromatographic method is developed for the separation and purification of four bioactive flavonoids from the medicinal herb Epimedium brevicornum Maxim. The crude flavonoids are obtained by extraction with ethyl acetate and ethanol from the dried aerial parts of Epimedium brevicornum Maxim under sonication. High-speed counter-current chromatography with a two-phase solvent system composed of n-butanol-ethyl acetate-water (3:7:10, v/v) is performed using a dual-mode method, which yields 3.2 mg of epimedin A at a purity of 98.2%, 5.5 mg of epimedin B at a purity of 92.6%, 12.7 mg of epimedin C at a purity of 90.4%, and 42.9 mg of icariin at a purity of 96.8% based on the HPLC analysis. The recoveries are 95.2%, 89.4%, 91.1%, and 94.8%, respectively. The isolation of these flavonoids in adequate amounts makes them readily available for structure-activity relationship studies and for quality control of the herbal medicine.