HPLC同时测定复方金银花颗粒中绿原酸、连翘苷、黄芩苷、汉黄芩素、芹菜素的含量

Simultaneous Determination of Chlorogenic Acid,Forsythin,Baicalin,Wogonin and Apigenin in Compound Jinyinhua Granules by HPLC

目的：建立同时测定复方金银花颗粒中5种化学成分（绿原酸、连翘苷、黄芩苷、汉黄芩素、芹菜素）的高效液相色谱法.方法：采用Agilent-Eclipse XDB-C18色谱柱（4.6 mm×250 mm,5μm）.以甲醇（A）-0.2％磷酸溶液（B）为流动相进行梯度洗脱（0 - 10 min,25％ -53％A;10-16 min,53％ - 78％A）,检测波长278 nm,流速1mL·min-1,进样量为20μL,柱温25℃.结果：绿原酸、连翘苷、黄芩苷、汉黄芩素、芹菜素5种成分在25 min内基本分离.峰面积（Y）对进样量（X）的标准曲线分别为绿原酸Y=2 350.4X-137.27,r=0.999 6;连翘苷Y=1 161.7X＋ 18.474,r=0.999 6;黄芩苷Y=3 204.2X-423.45,r=0.998 8;汉黄芩苷Y=4 792.7X-93.15,r=0.998 5;芹菜素Y=1 434.1X-8.4132,r=0.999 3;加样回收率分别为：绿原酸99.47％（RSD 0.8％）;连翘苷98.26％ （RSD 0.9％）;黄芩苷100.4％ （RSD 1.9％）;汉黄芩素99.29％ （RSD 1.1％）;芹菜素98.79％ （RSD 1.1％）.结论：该方法操作简便,重复性好,分离效果好,可以作为复方金银花颗粒的质量控制.

Objective：A rapid and sensitive method for HPLC was established to simultaneouly determine chlorogenic acid,forsythin,baicalin,wogonin and apigenin.Method：Chromatographic separation was carried out on Agilent Eclipse XDB-C18 column （4.6 mm ×250 mm,5 μm） under gradient elution （0-0.01 min,A phase is 25％ ; 0.01-10 min,A phase is 25％-53％ ; 10-16 min,A phase is 53％-78％） with methanol （A phase）-water/0.2％ phosphate （B phase） as mobile phase at a flow rate of 1.00 mL ·min-1 to elute at 25 ℃.The injection volume was 20 μL and the detection was adopted UV detector at 278 nm.Result：These five components were seperated basically within 25 minutes with good linearity.The regression equations were Y =2 350.4X-137.27,r =0.999 6 （for chlorogenic acid）,Y=1 161.7X ＋ 18.474,r =0.999 6 （for forsythin）,Y=3 204.2X-423.45,r=0.998 8 （for baicalin）,Y=4 792.7X-93.15,r=0.998 5 （for wogonin）,Y=1 434.1X-8.413 2,r =0.999 3 （for apigenin）.The average recoveries were：chlorogenic acid 99.47％ （RSD0.8％）,forsythin 98.26％ （RSD 0.9％）,baicalin 100.4％ （RSD 1.9％）,wogonin 99.29％ （RSD 1.1％）,apigenin 98.79％ （RSD 1.1％）.Conclusion：The HPLC method has good accuracy,reliability,and repeatbility.It is suitable for compound Honeysuckle granules and traditonal Chinese medicines containing the same ingredients to improve their quality control.