HPLC法测定乌梢蛇中六种核苷类成分含量

HPLC Method for Simultaneous Determination of 6 Nucleosides in Zaocys dhumnades(Cantor)

目的建立同时测定乌梢蛇中尿嘧啶、胞苷、次黄嘌呤、腺嘌呤、肌苷和鸟苷6种核苷类成分的HPLC测定方法。方法采用RP-HPLC法,色谱柱Agilent TC-18(4.6 mm×250 mm,5μm);流动相:0.05 mol/L KH2PO4-乙腈(99:1);体积流量为:0.6 ml/min;柱温:25℃;检测波长为260 nm;进样量:10μl。结果测定了乌梢蛇药材种6种核酸的含量,尿嘧啶、胞苷、次黄嘌呤、腺嘌呤、肌苷和鸟苷6种核苷类成分分别在1.73~8.65、1.79~8.97、2.22~11.10、3.28~16.42、4.28~21.40、4.71~23.56μg内呈良好的线性关系(r≥0.999 5),平均回收率为99.47%、97.70%、99.54%、99.15%、99.10%、99.91%,RSD<2.00%。结论该方法简单、准确可靠、重现性良好,可用于乌梢蛇核苷类成分含量测定及质量评价。

Objective To establish an HPLC method for simultaneous determination of 6 different nucleosides（uracil, cytidine, hypoxanthine, adenine, inoside, guanosine） in Zaocys dhumnades（Cantor）. Method RP-HPLC is applied in this study, Agilent TC-18（4.6 mm × 250 mm, 5 μm） is used, the mobile phase is a 0.05 mol/L KH2 PO4-acetonitrile（99 ： 1）, the flow rate is 0.6 ml/min, the temperature of column is 25℃ and the UV detector is set at 260 nm. The injection volume is 10 μl. Result Six nucleosides from the Zaocys dhumnades（Cantor） are measured. The linear range of uracil, cytidine, hypoxanthine, adenine, inoside and guanosine is 1.73 ~ 8.65, 1.79 ~ 8.97, 2.22 ~ 11.10, 3.28 ~ 16.42, 4.28 ~ 21.40, 4.71 ~ 23.56 μg, r ≥ 0.999 5, which indicated the linearity of this method is all right. The average recovery of them are 99.47 %, 97.70 %, 99.54 %, 99.15 %, 99.10 %, 99.91 %, and the RSD2.00 %. Conclusion This method is simple, accurate and reliable and has good reproducibility. This method can be used for determination of the content of nucleotide in Zaocys dhumnades（Cantor）, and quality evaluation for Zaocys dhumnades（Cantor）.