HPLC Quantitative Analysis of Uracil and Three Nucleotides in Fritillariae Bulbus and the Application for Variety Discrimination

OBJECTIVE To establish an HPLC method 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. METHODS 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 ℃. RESULTS Uracil, uridine, granosine and adenosine were baseline separated. The method had good linearity. CONCLUSION 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.