RP-HPLC测定细梗胡枝子中3种黄酮类有效成分

Determination of Three Flavonoids in Lespedeza virgata by RP-HPLC

目的:建立一种高效液相色谱测定细梗胡枝子中槲皮素、芦丁、山柰酚含量的方法。方法:采用反相高效液相色谱法,Agilent 1100 ZORBAX Eclipse XDB-C18色谱柱(4.6 mm×150 mm,5μm),二极管阵列检测器(DAD),甲醇-0.2%磷酸溶液(60∶40)为流动相,检测波长251 nm,流速1.0 mL.min-1,柱温35℃。结果:槲皮素在0.210～4.19μg,芦丁在0.084～1.68μg,山奈酚在0.066～1.33μg,峰面积与进样量呈良好的线性关系;芦丁、槲皮素和山奈酚的加样回收率分别为99.48%,98.53%,99.33%,RSD分别为0.51%,1.35%,2.13%(n=5)。结论:该方法精密度高、重复性和分离效果较好,分析快速准确,适合于该药材及提取物中槲皮素、芦丁和山奈酚3种黄酮类含量的测定,可作为该药材质量控制的参考依据。

Objective：To establish a quantification method for determination of quercetin,rutoside and kaempferol in Lespedeza virgata by HPLC.Method：RP-HPLC was applied on the determination of quercetin,rutoside and kaempferol in L.virgata with C18 column（4.6 mm × 250 mm,5 μm）,DAD detector,and a mixture of acetonitrile-0.2% phosphoric acid（60∶ 40） as a mobile phase with the flow rate at 1.0 mL.min-1.Three flavonoids were detected at 251 nm,column temperature was set at 35 ℃.Result：Quercetin,rutoside and kaempferol were linear in the range of 0.210-4.19,0.084-1.68,0.066-1.33 μg.The average recovery of rutoside,quercetin and kaempferol was 99.48%,98.53% and 99.33% with RSD（n = 5） 0.51%,1.35% and 2.13% respectively.Conclution：The method is accurate and sensitive,which can be used rapidly for the content determinatinon of rutoside,quercetin and kaempferol from L.virgata.It can be applied to the quality control of this crude drug.