高效液相色谱法测定抗菌消炎片中绿原酸、黄芩苷、黄芩素和大黄酚的含量

High performance liquid chromatography in determination of chlorogenic acid,baicalin,emodin and chrysophanol contents in Kangjunxiaoyan tablet

目的:采用高效液相色谱法测定抗菌消炎片中绿原酸、黄芩苷、黄芩素和大黄酚的含量。方法:抗菌消炎片去包衣粉末用50%甲醇超声提取30min;色谱柱为Agilen Eclipse Plus-C18柱(4.6mm×250mm,5μm);流动相:A相为0.1%磷酸水溶液,B相为甲醇,C相为乙腈,梯度洗脱;流速:0.8ml/min;温度:30℃;进样量:10μl;检测波长:254nm。结果:绿原酸、黄芩苷、黄芩素和大黄酚4种成分在45min内基线分离。峰面积(Y)对浓度(X)的标准曲线分别为:绿原酸Y=12.89X-12.66,r=0.9999;黄芩苷Y=13.96X-1.013,r=1.0;黄芩素Y=35.82X-4.923,r=0.9999;大黄酚Y=44.16X-3.280,r=0.9999。方法学考察表明,日内及日间精密度、最低检测限均符合相关标准,加样回收率分别为(n=3):绿原酸101.5%(RSD=1.6%);黄芩苷103.4%(RSD=1.4%);黄芩素99.3%(RSD=2.0%);大黄酚98.1%(RSD=1.7%)。结论:采用高效液相色谱法同时测定抗菌消炎片中绿原酸、黄芩苷、黄芩素和大黄酚的含量,样品处理简便,测定结果准确,可用于该制剂的质量控制。

Objective：To establish an HPLC method for determination of chlorogenic acid,baicalin,emodin and chrysophanol contents in Kangjunxiaoyan tablet. Methods：After removing the coating,the powder of Kangjunxiaoyan tablet was extracted by 50% methanol for 30 min using the following condition：analytical column,Agilent Eclipse Plus-C18 column（4.6 mm×250 mm,5 μm）; mobile phase：A,0.1% （v/v） phosphoric acid-water,B,methanol,C,acetonitrile with gradient elution. The flow rate was 0.8 ml/min; the detection wavelength was set at 254 nm; the temperature of column was 30℃; and the injection volume was 10 μl. Results：The title compounds were separated at baseline within 45 min with good linear,with the regression being Y=12.89X-12.66,r=0.999 9 for chlorogenic acid,Y=13.96X-1.013,r=1.000 0 for baicalin,Y=35.82X-4.923,r=0.999 9 for emodin,and Y=44.16X-3.280,r=0.999 9 for chrysophanol. The intra-day and inter-day precisions and the limits of detection were all within the normal range. The recovery rates（n=3） were：chlorogenic acid 101.5%（RSD=1.6%）,baicalin 103.4%（RSD=1.4%）,emodin 99.3%（RSD=2.0%）,and chrysophanol 98.1%（RSD=1.7%）. Conclusion：HPLC is a simple,accurate,stable,and reliable method in determining the contents of chlorogenic acid,baicalin,emodin,and chrysophanol in Kangjunxiaoyan tablet,and it can be used for quality control of this preparation.