Objective To establish a method for the determination of polydatin，deoxyrhapontin and piceatannol 3′-O-β-D-glucopyranoside by ultra performance liquid chromatography（UPLC）. Methods The assay was performed on a ACQUITY UPLC?誖 BEH C18 1.7μm 2.1×50mm Colum with acetonitrile-wayer（15：85） as mobile phase in gradient elution at a flow of 0.4 ml/min. The detection wavelength was set at 319nm. The colum temperature was 30 ℃. Results polydatin，deoxyrhapontin and piceatannol 3′-O-β-D-glucopyranoside showed good linearity in the ranges of 1.0225～204.5，1.02～204，1～200μg·mL-1，respectively. The average recoveries of polydatin was 101.35%（RSD=3.7），98.36%（RSD=1.81%） for deoxyrhapontin and 99.33（RSD=3.64%） for piceatannol 3′-O-β-D-glucopyranoside. Conclusion UPLC method may greatly improve the separation efficiency and analysis speed；This method can be applied in the determination of polydatin deoxyrhapontin and piceatannol 3′-O-β-D-glucopyranoside in Rheum Ihasaense.