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Simultaneous Determination of Five Nucleosides in Pheretima aspergillum (E. Perrier) by HPLC
GUAN Shuiqing, ZHOU Gailian, DONG Jingjing, LIN Qinghua, HUANG Pan, WANG Qian
Journal of Guangxi Normal University(Natural Science Edition). 2020, 38 (3):
85-91.
DOI: 10.16088/j.issn.1001-6600.2020.03.011
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).
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