高效液相色谱法测定五味黄连丸中表小檗碱、黄连碱、巴马汀和小檗碱的含量

Determination of epiberberine,coptisine,palmatine,and berberine in Wuwei Huanglian pills by high- performance liquid chromatography

目的：建立高效液相色谱法测定五味黄连丸中表小檗碱、黄连碱、巴马汀和小檗碱的含量。方法：采用Diamosil C18色谱柱（250mm×4.6mm,5μm）,流动相：乙腈-0.1%磷酸溶液（50∶50）（每100mL加十二烷基磺酸钠0.1g）;检测波长：345nm;流速：1.0mL/min;柱温：室温。结果：表小檗碱、黄连碱、巴马汀和小檗碱分别在0.489~11.73、0.992~23.81、1.014~24.35、2.017~48.40μg/mL峰面积与浓度之间呈良好线性关系;平均加样回收率分别为100.1%、98.3%、99.4%、99.4%,RSD分别为1.3%、1.7%、1.3%、2.8%。结论：所建立的HPLC法准确、快速、专属性强、灵敏度高,可用于五味黄连丸中表小檗碱、黄连碱、巴马汀和小檗碱的含量测定。

Objective ： To establish a method for determining the content of epiberberine, coptisine, palmatine, and berberine in Wu- wei Huanglian pills by high- performance liquid chromatography （HPLC）. Methods：HPLC was performed on a Diamonsil C18 column （250mm×4.6mm,5μm） with a mobile phase of acetonitrile -0. 1% phosphoric acid （50： 50,0. 1 g of sodium dodecyl sulfate was added per 100ml ） at a detection wavelength of 345 nm, a flow rate of 1.0 ml/min, and a column temperature of room temperature. Results ： Epiberberine, coptisine, palmatine, and berberine showed a good linear relationship between peak area and concen- tration within 0. 489 - 11.73,0. 992 -23.81,1. 014 - 24.35, and 2. 017 -48.40 μg/ml, respectively;the average recovery rates were 100. 1%, 98. 3%, 99.4%, and 99.4%, respectively, with relative standard deviations of 1.3%, 1.7%, 1.3%, and 2. 8%, respectively. Conclusion ：The established HPLC method is accurate and efficient and has high specificity and sensitivity and thus can be used for the determination of epiberberine, coptisine, palmatine, and berberine in Wuwei Huanglian pills.