Abstract: To establish an HPLC method for the quantitative of five nucleoside components in Pheretima aspergillum (E. Perrier).The mobile phase was eluted with Diamonsil C₁₈ (250 mm× 4.6 mm, 5 μm) by using methanol-water solution (5∶95) such as mobile phase with a flow rate of 1.0 mL/min and a detection wavelength of 254 nm. The column temperature was 30 ℃. The contents of 5 nucleoside components in P. aspergillum (E. Perrier) were determined. Uracil, hypoxanthine, uridine, inosine and guanosine showed a good linear relationship in the range of 14.84～111.30 ng (r=0.999 9), 126.00～945.00 ng (r=0.999 8), 63.60～477.00 ng (r=0.999 9), 612.00～4 590.00 ng (r=0.999 8) and 41.60～312.00 ng (r=0.999 9), with average recovery (n=9) of 99.27%, 99.14%, 98.59%, 98.69% and 99.91%, respectively. The RSD(n=9)was 0.95%, 0.86%, 1.01%, 0.86% and 1.91%, respectively. The contents of uracil, hypoxanthine, uridine, inosine and guanosine in 20 batches were 0.051 0~0.111 0, 0.243 1~0.943 6, 0.171 9~0.508 0, 1.559 6~3.861 1 and 0.165 5~0.250 1 mg/g, respectively. The content of 5 nucleosides was 3.309 9~4.565 9 mg/g. The established method was simple, reliable, reproducible and accurate, and can be used for the simultaneous determination of 5 nucleosides in P. aspergillum (E. Perrier).

Key words: Pheretima aspergillum (E. Perrier), nucleoside and mucleobase, content determination, HPLC