HPLC法同时测定复方金银花颗粒中10种成分的含量

Simultaneous assay of 10 active components in compound Jinyinhua granules by HPLC

目的建立HPLC法同时测定复方金银花颗粒中10种成分(新绿原酸、绿原酸、隐绿原酸、咖啡酸、连翘酯苷B、连翘酯苷A、黄芩苷、汉黄芩苷、黄芩素、汉黄芩素)含量的方法。方法采用HPLC法,色谱柱:Kromasil C_(18)(250 mm×4.6 mm,5μm);流动相:乙腈-0.1%磷酸水溶液,梯度洗脱;流速:0.8 ml/min;检测波长:320 nm;柱温:35℃;进样量:10μl。结果在上述色谱条件下,10个成分的峰具有良好的分离度;新绿原酸、绿原酸、隐绿原酸、咖啡酸、连翘酯苷B、连翘酯苷A、黄芩苷、汉黄芩苷、黄芩素、汉黄芩素的线性范围分别为10.20～340.06、7.19～239.81、6.12～203.84、15.00～500.00、20.16～671.85、25.19～839.67、14.61～487.14、10.52～350.66、7.32～244.01、7.61～253.82μg/ml;10个对照品的平均加样回收率均在97.67%～99.00%之间,RSD均在0.94%～1.37%之间;10批制剂中新绿原酸、绿原酸、隐绿原酸、咖啡酸、连翘酯苷B、连翘酯苷A、黄芩苷、汉黄芩苷、黄芩素、汉黄芩素的平均含量分别为0.579 0、1.523 5、1.080 9、0.083 1、0.468 0、0.739 6、1.977 9、0.351 5、0.246 8、0.256 6 mg/g。结论该含量测定方法简便、稳定可靠、准确,可作为复方金银花颗粒质量评价的方法。

Objective To establish an one step assay method for 10 active components(neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,forsythoside B,forsythoside A,baicalin,wogonoside,baicalein,wogonin)in compound Jinyinhua granules.Methods The Kromasil C 18 column(250 mm×4.6 mm,5μm)was used with the column temperature at 35℃and PDA detection wavelength at 320 nm.A gradient elution with acetonitri-0.1%phosphoric acid was adopted at the flow rate of 0.8 ml/min.The injection volume was 10μl.Results The good resolution was achieved for the 10 active components.The linear ranges of neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,forsythoside B,forsythoside A,baicalin,wogonoside,baicalein,wogonin were 10.20-340.06,7.19-239.81,6.12-203.84,15.00-500.00,20.16-671.85,25.19-839.67,14.61-487.14,10.52-350.66,7.32-244.01,7.61-253.82μg/ml respectively.The average recoveries(n=6)were between 97.67%and 99.00%and RSDs between 0.94%and 1.37%.The average contents of neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,forsythoside B,forsythoside A,baicalin,wogonoside,baicalein,wogonin from 10 batches of preparations were 0.579 0,1.523 5,1.080 9,0.083 1,0.468 0,0.739 6,1.977 9,0.351 5,0.246 8 and 0.256 6 mg/g.Conclusion This simple and accurate method can be used as one of quality control methods for the compound Jinyinhua granules.