Objective: To establish a high performance liquid chromatography method for simultaneous determination of eight nucleosides in Scolopendra. Method: Hedera ODS-2 C18 column (4.6 mm×250 mm

5 μm) was used for determination of eight nucleosides. The analyses were eluted with methanol and water in a gradient mode. The flow rate was 1.0 mL·min-1

and the column temperature was set at 30 ℃. The eight nucleosides were monitored and quantified at a wavelength of 254 nm

and the injection volume was 5 μL. Result: The investigated compounds

including cytosine

cytidine

hypoxanthine

xanthine

uridine

thymine

2'-deoxyinosine and thymidine

in the samples were well separated. The eight nucleosides showed a good linearity at a certain concentration (r>0.999 9)

and the mean recoveries of the eight nucleosides ranged from 95.73% to 102.6%. The contents of nucleosides in Scolopendra from different varieties varied significantly. The highest of content of the eight nucleosides was in Scolopendra mojiangica

and the second was in S. multidens. The content of the eight nucleosides in S. subspinipes mutilans and S. negrocapitis were close. The contents of hypoxanthine and xanthine in centipedes were significantly higher than those in other nucleosides

and the content of hypoxanthine in S. subspinipes mutilans was higher than that in other varieties. Conclusion: This high performance liquid chromatography method and quantitative analysis is simple

rapid

accurate and highly reproduced for simultaneous determination of eight nucleosides in Scolopendra

and helpful for its quality control.