RP-HPLC法测定乙肝宁颗粒中3种成分的含量

Determination of 3 components in Yiganning Granules by RP-HPLC

目的建立能同时测定乙肝宁颗粒中绿原酸、槲皮素和咖啡酸含量的RP-HPLC法。方法乙肝宁样品经超声提取后,采用Phenomenex SuperLu C 18(250 mm×4.6 mm,5μm)液相色谱柱;柱温:25℃;检测波长:341 nm;进样体积:20μL。流动相:乙腈(A)-3 mL·L^(-1)磷酸(B),梯度洗脱:0~5 min,7%A;5~26 min,7%A~65%A;26~28 min,65%A~7%A;28~35 min,7%A。结果3种待测成分质量浓度分别在3.872~387.200、2.991~299.100、0.399~39.920μg·mL^(-1)范围内线性关系良好,r 2均大于0.9990;平均回收率分别为100.72%、100.15%、99.71%;重复性和精密度RSD值(n=6)均小于5.0%。结论该方法操作简单、准确度高、重复性好,适用于乙肝宁颗粒中3种成分的含量测定。

Objective To establish an analytical method for the determination of chlorogenic acid,quercetin and caffeic acid in Yiganning Granules by RP-HPLC.Methods Yiganning Granules samples were treated by ultrasonic extraction.Phenomenex SuperLu C 18(250 mm×4.6 mm,5μm)was selected as chromatographic column.The column temperature was 25℃.The detection wavelength was 341 nm and sample injection volume was 20μL.Acetonitrile(A)-3 mL·L^(-1)phosphoric acid(B)was used as the mobile phase in a gradient elution mode.The gradient elution procedure was as follows:0-5 min 7%A,5-26 min 7%A-65%A,26-28 min 65%A-7%A,28-35 min 7%A.Results The 3 components showed good linear relationships in the concentration ranges of 3.872-387.200,2.991-299.100 and 0.399-39.920μg·mL^(-1),with correlation coefficients(r 2)all above 0.9990.The average recoveries were 100.72%,100.15%and 99.71%,respectively with RSD of repeatability and precision less than 5.0%(n=6).Conclusion The method is simple,accurate and reproducible,which is suitable for the determination of the 3 components in Yiganning Granules.