[Objective] To develop reverse phase high performance liquid chromatography(RP-HPLC) method for simultaneous determination of 9 nucleosides and nucleobases in Fritillaria anhuiensis S.C.Chen et S.E.Yin.[Methods] The analyses were performed on an Agilent Zorbax SB-Aq C18(4.6 mm × 250 mm,5 μm)for all analyses.The investigated compounds were separated with gradient mobile phase consisting of water and Methanol.The flow rate was 1.0 ml/min.The detection wavelength was 260 nm with the 25 ℃ temperature of column.[Result] The investigated compounds including uracil,cytidine,uridine,hypoxanthine,inosine,guanosine,thymidine,adenosine,adenine had good linearity(r2 ≥ 0.998 2).The average recovery was 96.7% to 104.3% with RSD ≤ 3.87%.[Conclusion] The accuracy,stability,repeatability,and average recovery of the method are satisfying,and the 9 nucleosides and nucleobases of F.anhuiensis can be rapidly and accurately quantified by RP-HPLC. [Objective] The developed reverse phase high performance liquid chromatography (RP-HPLC) method for simultaneous determination of 9 nucleosides and nucleobases in Fritillaria anhuiensis SC Chen et SEY in. [Methods] The analyzes were performed on an Agilent Zorbax SB-Aq C18 4.6 mm × 250 mm, 5 μm) for all analyzes. The research compounds were separated with gradient mobile phase consisting of water and Methanol. The flow rate was 1.0 ml / min. The detection wavelength was 260 nm with the 25 ° C temperature of column . [Result] The investigated compounds include uracil, cytidine, uridine, hypoxanthine, inosine, guanosine, thymidine, adenosine, adenine had good linearity (r2 ≥ 0.998 2). The average recovery was 96.7% to 104.3% with RSD ≤ 3.87%. [Accuracy] The accuracy, stability, repeatability, and average recovery of the method are satisfied, and the 9 nucleosides and nucleobases of F. cannensis can be rapidly and accurately quantified by RP-HPLC.