In combination with the secondary metabolism and high resolution mass spectrometry of the flavanol components of Epimedium,a biogenic synthesis pathway with wushanicariin as the characteristic product was constructed to derive and verify the chemical structure of new components. Fifty-four batches of Epimedium samples from 15 varieties were collected by high resolution mass spectrometry. The chemical structure of the deduced metabolites was analyzed and verified by using the functions of Formula Finder,Mass Calculators,XIC manager,online database (SciFinder,Reaxys,ChemSpider,etc.) and the fragmentation rule of secondary mass spectrometry of PeakView software. In this experiment,12 new compounds were identified in different kinds of Epimedium samples. This research method can simplify the complicated steps of phytochemical separation and provide a new way for the analysis and identification of bioactive secondary metabolites.