HPLC法同时测定柴胡—黄芩药对水煎液中皂苷和黄酮类的含量

Simultaneous determination of saponins and flavonoids decoctions of bupleutrum chinese-scutellaria baicalensis Georgi by HPLC

目的建立高效液相色谱测定柴胡—黄芩药对水煎液中皂苷和黄酮类含量的方法。方法采用MERCK PUROSPHERC18柱(250 mm×4.6 mm,5μm);流动相:甲醇—0.05%磷酸,梯度洗脱;流速:1.0 mL.min-1;柱温:35℃;检测波长为210 nm。结果皂苷成分(柴胡皂苷a)、3个黄酮类成分(黄芩苷、黄芩素、汉黄芩素)达到了完全分离,4个成分的峰面积与进样量线性关系良好。柴胡皂苷a、黄芩苷、黄芩素、汉黄芩素线性范围分别为0.219 6～17.568μg、0.214 75～17.18μg、0.093 1～7.448μg、0.107 6～8.608μg,(r≥0.999 8);平均回收率分别为97.71%、102.43%、101.16%、97.50%;RSD分别为0.52%、1.96%、1.55%、0.75%。结论该方法可同时测定柴胡—黄芩药对水煎液中皂苷和黄酮类成分的含量,并能有效表征水煎液中其他成分差别,也为建立柴胡—黄芩药对水煎液特征指纹图谱奠定了基础。

Objective To establish a high performance liquid chromatographic determination method for saponins and flavonoids of bupleutrum chinese-scutellaria baicalensis Georgi.Methods The MERCK PUROSPHER C18 column（250 mm × 4.6mm,5 μm） was used,with a mobile phase of methanol-0.05% phosphate,Gradient elution.The flow rate was 1 mL.min-1.The column temperature was set at 35℃.The detection wavelength was 210nm.Results Saponins（Saikosaponin a）,3 Flavonoids（Baicalin,Baicalein,Wogonin） achieved a complete separation,Peak area of the 4 components had a good linear relationship with the amount of samples.The linear ranges of Saikosaponin a,Baicalin,Baicalein,and Wogonin were 0.219 6 ～ 17.568 μg,0.214 75 ～ 17.18 μg,0.093 1 ～ 7.448 μg,0.107 6 ～ 8.608 μg,respectively（r≥0.999 8）;The average recoveries were 97.71%,102.43%,101.16%,97.50%.RSD were 0.52%,1.96%,1.55%,0.75%.Conclusion This method can be used to determine saponins and flavonoids of bupleutrum chinese-scutellaria baicalensis Georgi medicine,and can effectively characterize the differences among the other ingredients in the decoction,but also lay a foundation for the establishment of fingerprint of bupleutrum chinese-scutellaria baicalensis Georgi.