An HPLC method was established for simultaneously determination of uracil, uridine, granosine and adenosine in bulbus of F. thunbergii Miq., F. cirrhosa D. Don, F. ussuriensis Maxin, and F. walujewii Regel, and for identification of these varieties. The HPLC method was used with Alltima C18 column (4.6 mm × 250 mm, 5 μm). The gradient elution solvent system was MeOH/H2O with time program as follows: 0.0 min→5.0 min→10.0 min→20.0 min→30.0 min→40.0 min, 1%→1%→5%→12%→24%→31% (MeOH). The UV wavelength was 260 nm; flow rate was 0.8 mL · min-1; column temperature was 25°C. Uracil, uridine, granosine and adenosine were baseline separated The method had good linearity. The method is rapid, precise and repeatable, and can be used for the determination of nucleosides and supplying evidence for variety discrimination of bulbus of F. thunbergii Miq., F. cirrhosa D. Don, F. ussuriensis Maxin, and F. walujewii Regel.