OBJECTIVE：To establish a method for the content determination of diosgenin in Dioscorea zingiberensis，and to compare the contents of diosgenin in wild D. zingiberensis from different habitats. METHODS：HPLC method was adopted. The determination was performed on Inertsil ODS-3 column with mobile phase consisted of acetonitrile-water （50 ∶ 50，V/V） at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃. The detection wavelength was set at 203 nm，and sample size was 10 μL. Established method was used to determine the contents of diosgenin in wild D. zingiberensis from different habitats （Shaanxi Shangluo，Shaanxi Ankang，Henan Neixiang，Yunnan Xuanwei，Sichuan Deyang，Hubei Shiyan，Hunan Zhangjiajie，Hunan Changde）. The differences of diosgenin content were compared. RESULTS：The linear range of diosgenin were 4.16-165.6 μg/mL （r＝     0.999 9）. RSDs of precision，reproducible and stability tests were all lower than 2％ （n＝5 or n＝6）. The average recovery of diosgenin was 101.18％ （RSD＝1.27％，n＝6）. The content of diosgenin in wild D. zingiberensis from different habitats were in descending order，i.e. Sichuan Deyang （1 405.36 μg/g）＞Shaanxi Shangluo （1 201.79 μg/g）＞Hunan Zhangjiajie （1 035.18 μg/g）＞Shaanxi Ankang （632.64 μg/g）＞Hunan Changde （598.64 μg/g）＞Yunnan Xuanwei （425.34 μg/g）＞Henan Neixiang （350.13         μg/g）＞Hubei Shiyan （338.39 μg/g）. CONCLUSIONS：Established method is simple，accurate and suitable for the content determination of diosgenin in D. zingiberensis. The contents of diosgenin in wild D. zingiberensis from different habitats are different significantly，and the content of diosgenin in the samples from Sichuan Deyang is the highest.