OBJECTIVE： To explore the contents of secoiridoid substances （gentiopicroside， swertiamarin， sweroside） in the leaves of Gentiana rigescens from different populations in Dali Bai Autonomous Prefecture （Shorted for Dali prefecture）， and to provide reference for the selection of fine varieties， the development and utilization of G. rigescens. METHODS： The contents of secoiridoid substances in the leaves of G. rigescens from different populations in Dali prefecture were determined by HPLC， and the HPLC fingerprint was established. The contents of secoiridoid substances in the leaves of G. rigescens from different populations was studied by using single factor variance analysis， clustering analysis and principal component analysis. RESULTS： There were statistical significance in the contents of gentiamarin， swertiamarin and sweroside and the total contents in the leaves of G. rigescens from different populations in Dali prefecture （P＜0.05 or P＜0.01）. Twelve common peaks were found in HPLC fingerprint， and HPLC fingerprint similarities of common peaks except Gantong population were all over 0.972. Three peaks of them were identified as the characteristic peaks of gentiamarin， swertiamarin and sweroside which were correlated to the activities. The total contents of gentiamarin， swertiamarin and sweroside from different populations obtained by clustering analysis were consistent with that by HPLC fingerprint principal component analysis. CONCLUSIONS： There are great differences in the content of gentiamarin， swertiamarin and sweroside in the leaves of G. rigescens from different populations in Dali prefecture. The similarity of secoiridoid substances in the leaves of G. rigescens from different populations except Gantong population is higher.