HPLC法测定双黄连口服液中绿原酸和黄芩苷的含量

Determination of Chlorogenic Acid and Baicalin In Shuanghuanglian Koufuye by HPLC

目的建立HPLC法同时测定双黄连口服液中绿原酸和黄芩苷含量的方法。方法色谱柱用十八烷基硅烷键合硅胶为填充剂(AgilentZORBAXEclipseXDB-C18,4.6mm×150mm,5Micron)。流动相A:甲醇-水-磷酸(50∶950∶5),流动相B:甲醇。梯度洗脱程序:0～30,A:95%～35%,B:5%～65%;流速:1ml/min,检测波长327nm。结果绿原酸和黄芩苷的保留时间分别约为10.6、24min,与各自相邻峰的分离度均>1.5。以峰面积(X)对进样浓度(Y)线性回归,绿原酸回归方程:Y=0.03298X-0.09386,r=0.9999,线性范围5.040～45.36μg.ml-1。黄芩苷回归方程:Y=0.05155X-1.2864,r=0.9999线性范围:44.88～403.9μg.ml-1。绿原酸、黄芩苷回收率(n=6)分别为99.6%和100.0%、RSD分别为0.32%和1.0%。结论本方法与2005版中国药典法含量测定结果一致,准确度好且操作简便。

Ahn To develop an HPLC quantitative method for the determination of chlorgenic acid and baicalin in Shuanghuanglian Koufuye. Methods The chromatographic conditions include the column C18 （Agilent ZORBAX Eclipse XDB-C18 ,4. 6mm × 150mm,SMicron） mobile phase -A methanol-water-phosphore acid （50： 950：5 ）, B methanol ;gradient elution ：0- 30,A ：95% -35%, B ：5% -65%. The flow rate was 1.0ml/min and monitored at 327nm. Results the retention time of chlorgenic acid and baicalin was 10.6rain and 24rain respectively. The regress equation for chlorgenic acid was Y =0. 032 98X -0. 093 86, r =0. 999 9,and the linear range was 5.040 -45.361xg·ml^-1. Baicalin was Y =0. 051 55X - 1. 286 4, r =0. 999 9 and the linear range was 44. 88 -403.91.tg·ml^-1. The average recovery of chlorgenic acid and baicalin was （ n =6）99.6% and 100.0% ; RSD 0. 32% and 1.0% respectively. Conclusion This method conform to the method of determination specified in CP 2005 ,and was sensiteve, time-saving and accurate.