Abstract: OBJECTIVE To establish fingerprint of Anemarrhenae Rhizoma UPLC. METHODS The UPLC fingerprint of Anemarrhenae Rhizoma were determined on an HSS T3 column (2.1 mm×100 mm,1.8 μm) eluted with the mobile phase consisted of acetonitrile and 0.03% phosphoric acid in gradient mode; flow rate: 0.5 mL·min^-1; column temperature: 30 ℃ and the detection wavelength was set at 210 nm. RESULTS The common mode of the UPLC fingerprint was set up under the established condition. There were 12 common peaks in the fingerprint of 16 samples, six of which were identified. The Anemarrhenae Rhizoma from 16 different areas could be divided into 2 grades through the results of hierarchical cluster analysis. CONCLUSION The method was fast and accurate. The chromatographic profile of Anemarrhenae Rhizoma with high specificity can be used to control the quality of Anemarrhenae Rhizoma.