HPLC法监测金英黄归汤制备过程中绿原酸和连翘酯苷A含量

Monitoring of Chlorogenic Acid and Forsythoside A Contents During Process of Jinying Huanggui Decoction by HPLC

目的采用高效液相色谱(HPLC)法建立绿原酸和连翘酯苷A的含量测定方法,并对制备过程中绿原酸和连翘酯苷A含量进行监测,为金英黄归汤制备工艺改进提供质控标准。方法采用Hypersil C18色谱柱(150mm伊4.6 mm,5μm);绿原酸和连翘酯苷A的流动相分别为乙腈-0.2%磷酸(11颐89)与乙腈-0.4%冰醋酸(17颐83);检测波长分别为327 nm和330 nm;柱温分别为35益和30益;流速分别为0.8 m·min-1和1.0 m L·min-1。结果绿原酸和连翘酯苷A的线性范围分别为:13.56耀217.00μg·m L-1(r2=0.9997),0.0536耀0.2680 mg·m L-1(r2=0.9996);稳定性试验RSD分别为0.14%和0.47%;中间精密度试验RSD分别为0.63%和0.56%;重复性试验RSD分别为0.68%和1.00%;平均回收率分别为绿原酸99.23%,RSD为0.61%;连翘酯苷A100.24%,RSD为1.15%。结论建立的绿原酸和连翘酯苷A含量检测方法,可为金英黄归汤制备过程提供质控数据。

Objective To establish the determination method for chlorogenic acid and forsythoside A in Jinying Huanggui decoction by HPLC, thus to provide the quality control standard for the improved product technology of Jinying Huanggui decoction. Methods Hypersil C18 column（150 mm＆#215; 4.6 mm,5 μm）was used. The mobile phase of chlorogenic acid was acetonitrile-0.2% phosphonic acid solution（11∶89）and that of forsythoside A was acetonitrile-0.4 % glacial acetic acid（17∶83）. The detection wavelength for chlorogenic acid and forsythoside A was set at 327 nm and 330 nm,the column temperature was 35℃and 30℃,and the flow rate was 0.8 mL·min^-1 and 1.0 mL·min^-1, respectively. Results The linear range of chlorogenic acid and forsythoside A in the established method was 13.56~217.00 μg·mL^-1（r2=0.9997）and 0.0536~0.2680 mg·mL^-1（r2=0.9996）, RSD of stability test was 0.14% and 0.47%, RSD of intermedial precision test was 0.63% and 0.56%, and RSD of the repeatability test was 0.68% and 1.00%, respectively. The average recovery of chlorogenic acid was 99.23%,RSD being 0.61%,and that of forsythoside A was 100.24%,RSD being 1.15%. Conclusion The determination method for chlorogenic acid and forsythoside A has been established, and the method will provide evidence for the quality control of Jinying Huanggui decoction preparation process.