AbstractEucommia ulmoides Oliver is a dioecious plant, which plays an important role in traditional Chinese medicine. However, there has not yet been any research on male and female E. ulmoides. The UPLC fingerprints and OPLS‐DA approach were able to quickly and easily identify and quantify E. ulmoides and differentiate between the male and female fingerprints. In this study, we optimized the UPLC conditions and analyzed them to investigate fingerprints of twenty‐four extracts of Eucommiae Cortex (EC) and twenty‐four extracts of Eucommiae Folium (EF) under optimal conditions. It was demonstrated that thirteen and twelve substances were possible chemical markers for EC and EF male and female discrimination and that the level of these markers – chlorogenic acid and protocatechuic acid – was many times higher in male than in female. This approach offered a reference for quality control and precise treatment of male and female E. ulmoides in the clinic.