Derivation of the biogenic pathway and identification of new compounds of Epimedium based on Yinyanghuo as the main metabolite

The chemical structures of new components of Epimedium were deduced and verified by combining the secondary metabolism of Epimedium flavonoids with high resolution mass spectrometry. Based on the literature of Epimedium chemistry, the biosynthesis pathway of Yinyanghuo was constructed, and the possible metabolites were deduced. This metabolite information was entered into the PeakView software program and the ions meeting the quality error of less than 5 ppm with correct isotope distribution and containing secondary fragments were taken as the target compounds. Through the use of the software Formula Finder, Mass Calculators, online database (ChemSpider, Metlin, HMDB, etc.) and the fragmentation law of secondary fragments, the chemical structures of 22 metabolites were determined. One new component and eight new compounds were identified in 54 batches of Epimedium samples from 15 varieties by high resolution mass spectrometry. This study avoids the long time and tedious steps of phytochemical separation, saves experimental costs, and provides a new method for the analysis and identification of secondary metabolites with pharmacodynamic activity.