Epimedium extracts was roughly separated by macroporous resin, and screened by mouse sleep aid and autonomic activity model. The active separation component is further screened by MCI and ODS chromatography column to separate the effective components to obtain 4 monomer ingredients. Identified by nuclear magnetic resonance structure, the four monomers were: epimedin C, epimedin B, icariin, icarisid. The results showed that epimedin C and icariin effectively increased the rate of sleep in mice, prolonged the sleep time, and reduced the number of activities and standing times. And the effect of epimedin C is better. The main functional component of epimedium with improved sleep is epimedin C, and icariin also has a good sleep aid function.