HPLC同时测定黄芪葛根汤中9种异黄酮类有效成分  

Simultaneous determination of nine isoflavonoids in Huangqi Gegen decoction by high performance liquid chromatography

作　　者：王治平[1] 李卫民[1] 高英[2] 刘杰[1] 

机构地区：[1]广州中医药大学中药学院,广州510006 [2]广州中医药大学新药研究开发中心,广州510006

出　　处：《中国中药杂志》2010年第20期2689-2692,共4页China Journal of Chinese Materia Medica

摘　　要：目的：建立黄芪葛根汤中9种异黄酮类有效成分的HPLC-UV测定方法。方法：采用HPLC法,Kromasil 100-5C18色谱柱（4.6 mm×250 mm,5μm）,Phenomenex C18保护柱（4.0 mm×3.0 mm）;流动相甲醇-0.1%枸橼酸溶液,梯度洗脱;流速1.0 mL.min-1,柱温25℃,检测波长250 nm。结果：9种异黄酮类成分的线性范围分别为3′-羟基葛根素81.28406.4ng、葛根素670.4335 2 ng、大豆苷136.96684.8 ng、毛蕊异黄酮苷33.376166.88 ng、染料木苷22.848114.24 ng、芒柄花苷26.176130.88 ng、大豆苷元31.04155.2 ng、毛蕊异黄酮44.544222.72 ng、芒柄花素26.56132.8 ng,相关系数（R2）均大于0.999 6,平均加样回收率分别为99.27%,102.4%,98.46%,103.1%,101.3%,99.71%,102.28%,97.89%,100.8%,其RSD分别为1.8%,2.0%,1.4%,1.4%,1.3%,1.8%,1.3%,0.97%,1.8%。结论：该法简便快速,重复性良好,结果准确可靠,可用于黄芪葛根汤中主要异黄酮类成分的含量测定。Objective： To develop a reversed-phase high performance liquid chromatographic method（RP-HPLC） for the simultaneous analysis of nine isoflavonoids in Huangqi Gegen decoction.Method： HPLC determination was performed on a Kromasil C18 column（4.6 mm×250 mm,5 μm）and detected at 250 nm.The mobile phase was consisted of methanol and 0.1%citric acid solution with gradient elution.Flow-rate was 1.0 mL·min-1,and column temperature was 25 ℃.Result： The method was proved to be linear in the ranges of 81.28-406.4,670.4-3 352,136.96-684.8,33.376-166.88,22.848-114.24,10.176-50.88,31.04-155.2,44.544-222.72,26.56-132.8 ng for the nine isoflavonoid,3′-hydroxypuerarin,puerarin,daidzin,calycosin-7-O-β-D-glucoside,genistin,ononin,daidzein,calycosin and formononetin respectively.The average recoveries were 99.27%,102.38%,98.46%,103.06%,101.29%,99.71%,102.28%,97.89% and 100.78%,respectively,RSD was 1.8%,2.0%,1.4%,1.4%,1.3%,1.8%,1.3%,0.97% and 1.8%,respectively.Conclusion： Results obtained show that it′s a convenient,accurate and reliable method.The method has been successfully applied to the simultaneous determination of the nine isoflavonoids in Huangqi Gegen decoction.

关 键 词：黄芪葛根汤 有效成分 异黄酮 高效液相色谱法 对药 

分 类 号：R284.1[医药卫生—中药学]