Quality evaluation of Epmedii Folium from different species based on spectrum-efficacy relationship

Epimedii Folium (EF) is a widely used traditional Chinese drug that encompasses a variety of species. It has been reported that the active constituents in EF vary, leading to the uneven quality of commercial medicinal materials. To investigate the specific differences, we developed a comprehensive evaluation method to access the quality of EF from four species. First, UPLC-Triple TOF-MS/MS was used to generate characteristic fingerprints of Epimedium samples; second, adenine-induced Kidney Yang deficiency model was established to evaluate the quality of four Epimedium varieties by evaluating biochemical markers and morphology; third, multivariate statistical analysis, including gray correlation analysis and bivariate correlation analysis was combined; and finally, UPLC-QTRAP MS/MS identified a potential biomarker. The results showed that 12 common peaks were identified in 40 batches derived from four Epimedium species. The severity of kidney and testicular lesions in experimental groups of rats showed significant improvement compared with the model. GCA and BCA indicated that three ingredients, icariin, quercitrin, and epimedin B were potential biomarkers, confirmed using LC-MS. In addition, epimedin B and icariin were significantly higher in EBM compared to the other three species, consistent with the pharmacological tests. The quality and efficacy of EF from different origins were stable, and all of them had protective effects on Kidney Yang deficiency of rats. Especially, all data suggested that EBM possesses superior quality than the other three. Overall, our work offers fundamental data for the thorough assessment and a fresh viewpoint on the quality control of EF from several species.