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ObjectiveThe technical system of the simultaneous determination of polydatin and resveratrol in Polygonum cuspidatum was established and the content of the two ingredients in the Polygonum cuspidatum from 5 producing areas in southwest China was compared. MethodThe two ingredients could be determined with HPLC method at once. The column was Agilent C 18 ( 150 mm × 4.6 mm,5 μm) ; the mobile phase,acetonitrile water ( 23∶77,V/V) and the detection wavelength,306 nm. ResultsThe injection concentration of polydatin and resveratrol was in the range of 48 480 μg/mL ( r = 0. 999 0) or 7. 2 72. 0 μg/mL ( r = 0. 999 2) ,respectively,which integral value of each peak area showed a good linear relationship and the average recovery rate of two components was 98. 69% and 99. 42% ,RSD was 1. 68% and 3. 39% ( n = 6) . The order of the content of polydatin in Polygonum cuspidatum from five producing areas was Leshan in Sichuan > Dujiangyan in Sichuan,Yongchuan in Chongqing > Zunyi and Bijie in Guizhou; the content of resveratrol,Zunyi and Bijie in Guizhou > Dujiangyan and Leshan in Sichuan,Yongchuan in Chongqing. Conclusion This test method can be used for the quality control of Polygonum cuspidatum,and the test results would provide a reference for the research on the medicinal function of Polygonum cuspidatum.