A comprehensive analytical method based on ultra performance liquid chromatography-quadrupole linear ion trap mass spectrometry（UPLC-QTRAP-MS/MS） was developed for the simultaneous determination of 10 nucleosides and nucleobases（uracil,cytidine,guanine,uridine,adenine,inosine,guanosine,thymidine,adenosine,2’deoxycytidine）in Polygoni Multiflori Radix.The sample was extracted with ultrapure water by ultrasonic machine at room temperature and centrifuged.Under the optimized conditions,good separations for 10 target compounds were obtained on a Waters Atlantis T3 column（2.1 mm×150 mm，3 μm）with a mobile phase of methanol-5 mmol/L ammonium acetate（0.1% acetic acid） at a flow rate of 0.4 mL/min.The target compounds were analyzed in the positive ion multiple reaction monitoring（MRM） mode.As a result,10 nucleosides and nucleobases obtained good linearities in the certain concentration ranges with correlation coefficients more than 0.99,and the limits of detection were in the range of 1.05-9.68 ng/mL.Average recoveries of 10 compounds were between 97.8% and 104.8%,and the relative standard deviations were between 1.8% and 5.0%.The method was simple,sensitive and accurate,and could provide a reliable and effective technique for the quality control of Polygoni Multiflori Radix.