Abstract: OBJECTIVE To establish HPLC fingerprint of Rheum palmatum from Gansu and provide a reliable method for its scientific evaluation and quality control.METHODS The main components were separated on a Agela Venusil XBP-C₁₈ (4.6 mm×250 mm， 5 μm) column and detected at 254 nm by gradient elution at the flow rate of 1.0 mL/min using acetonitrile-0.1% phosphoric acid as the mobile phase. Column temperature was at 40 ℃.RESULTS The main components of the samples harvested from 8 locations at Li County of Gansu were successfully and effectively separated and 26 peaks could be found from all chromatograms. 5 major peaks were identified based on the comparison with reference substances.CONCLUSION The method can be used for quality control and provide a valuable reference for Good Agriculture Practice of Rheum palmatum.