RP-HPLC同时快速测定鹿茸中尿嘧啶、次黄嘌呤、尿苷含量

RP-HPLC simultaneous determination of three biological base in antler velvet

目的:建立反相高效液相色谱法同时快速测定鹿茸药材中尿嘧啶、次黄嘌呤、尿苷3种生物碱基成分含量。方法:RP-HPLC外标法。采用Agilent TC C18色谱柱(250mm×4.6mm,5μm),流动相为含0.07%醋酸的3%甲醇溶液,流速1.0mL·min-1,检测波长254nm,柱温25℃。结果:在上述色谱条件下,尿嘧啶、次黄嘌呤和尿苷能够得到很好的分离;浓度均在1.7～33.3mg·L-1的范围内与其峰面积线性关系良好,相关系数分别为0.9995,0.9999,0.9996;最低检测限浓度(S/N=3)分别为9.375,18.75,75μg·L-1;平均加样回收率(n=3)在98.4%～103.4%范围内。结论:本方法分离度好,简便快速,重复性好,可用于鹿茸药材中尿嘧啶、次黄嘌呤、尿苷3种生物碱基成分的同时快速测定,具有很高的实用价值。

Objective：To establish an HPLC method for simultaneous and rapid determination of three main biological base components including uracil,hypoxanthine and uridine in antler velvet.Methods：An external standard method with Agilent TC C18 column（250 mm×4.6 mm,5 μm） was adopted with the mobile phase was consisted of 3% methanol-water solution containing 0.07% acetic acid at the flow rate of 1.0 mL·min-1 The detection wavelength was 254 nm and the column was 25 ℃.Results：Uracil,hypoxanthine and uridine were well separated by this method.Linearities of uracil,hypoxanthine and uridine were good（r=0.9995,0.9999,0.9996）in the range of 1.7-33.3 mg·L^-1.The detection limits（S/N=3） were 9.375,18.75 and 75 μg·L^-1 for uracil,hypoxanthine and uridine,respectively.The average recoveries of the three biological base components were found in the range of 98.4%^-103.4%.Conclusion：The method is not only simple,rapid and presents good resolution and reproducibility,but also shows a high utility value,which has been successfully applied to the simultaneous determination of uracil,hypoxanthine and uridine in antler velvet.