HPLC梯度洗脱法同时测定银黄软胶囊中绿原酸和黄芩苷的含量

Simultaneous determination of chlorogenic acid and baicalin in Yinhuang soft capsule by HPLC with gradient elution

目的:建立银黄软胶囊中绿原酸和黄芩苷含量同时测定的高效液相色谱法。方法:采用Symmetry-C_(18)柱(4.6mm×250 mm,5μm),乙腈-0.4%磷酸水溶液为流动相梯度洗脱,检测波长319 nm,柱温35℃,流速1.0 mL·min-1,进样量10μL。结果:绿原酸在范围1.67μg·mL^(-1)~16.7μg·mL^(-1)内呈良好的线性关系,平均回收率为99%,RSD为2.1%(n=6);黄芩苷在范围3.32μg·mL^(-1)~33.2μg·mL^(-1)内呈良好的线性关系,平均回收率为97%,RSD为1.1%(n=6)。结论:本实验所建立的色谱分析方法能够简单、快速、准确地测定银黄软胶囊中黄芩苷和绿原酸的含量。不同厂家的银黄软胶囊中绿原酸和黄芩苷含有量具有显著差异,建立黄芩苷和绿原酸的定量控制方法具有重要意义。

Objective： To establish a method for the simultaneous determination of chlorogenic acid and baicalin in Yinhuang soft capsule. Method： The chromatographic separations were obtained on a Symmetry-C18 column （250 mm×4.6 mm, 5μm）. The mobile phase was consisted of acetonitrile and 0.4% phosphoric acid solution in a gradient elution manner with a flow rate of 1.0 mL.min-1 at 35 ℃. The detection wavelength was set at 319 nm. Sample volume was 10μL. Result： Linear range of chlorogenic acid was 1.67- 16.7μg. mL-1 and that of baicalin was 3.32 -33.2 μg-mL1. The recovery of chlorogenic acid and baicalin was 99.0% （RSD=2.1%） and 97% （RSD=1.1%）, respectively. Conclusion： The chromatographic method established can be used simply and accurately to detect the content of chlorogenic acid and baicalin in Yinhuang soft capsule. The contents of chlorogenic acid and baicalin are different in different manufacturer productions, and it is necessary to establish the quantitative control of chlorogenic acid and baicalin.