HPLC法测定新疆阿尔泰锦鸡儿根、茎、叶、花中3种黄酮类化合物的含量

Content determination of three flavonoids in roots,stems,leaves and flowers of Caragana altaica Kom.Pojark by HPLC method

目的建立高效液相色谱法(HPLC法)测定新疆阿尔泰锦鸡儿根、茎、叶、花中槲皮素、山奈酚、异鼠李素含量的方法。方法将阿尔泰锦鸡儿根、茎、叶、花的75%乙醇提取物经酸水解处理制备成供试品溶液,采用HPLC法:Inertsil ODS-3色谱柱(150mm×4.6mm,5μm);流动相为甲醇-0.4%磷酸(48∶52),进样量10μL,柱温25℃,流速1.0mL/min,检测波长360nm。测定槲皮素、山奈酚、异鼠李素的含量。结果槲皮素、山奈酚、异鼠李素分别在6.16~123.14、3.99~79.76、2.40~48.10μg/mL浓度范围内线性关系良好,平均回收率分别为96.40%、96.95%、93.48%,RSD分别为3.5%、3.7%、3.7%。阿尔泰锦鸡儿花中槲皮素、山奈酚、异鼠李素的平均含量分别为0.576 1、0.265 0、0.173 3mg/g,叶中槲皮素、山奈酚、异鼠李素的平均含量分别为1.502 9、1.345 8、0.450 4mg/g,茎中槲皮素的平均含量为0.128 0mg/g,未检出山奈酚和异鼠李素,根中均未检出这3种化合物。结论本研究建立的方法简便、稳定性、重复性良好,适用于新疆阿尔泰锦鸡儿药材中黄酮类化合物的含量测定。

Objective To establish the method for content determination of three main flavonoids,quercetin,kaempferol and isorhamnetin,in Caragana altaica Kom.Pojark.Methods The 75% ethanol extracts from roots,stems,leaves and flowers of Caragana altaica Kom.Pojark were prepared by acid hydrolysis as sample solution,followed by analysis of sample solution were performed on Inertsil ODS-3 chromatographic column（150mm×4.6 mm,5μm）,with the mobile phase consisting of methanol-0.4% hosphoric acid（48∶52）at the flow rate of 1.0mL/min.UV detection wavelength was set at 360 nm and the column temperature was 25℃.The content of quercetin,kaempferol and isorhamnetin was determined.Results There was good linear relationship found within 6.16~123.14,3.99~79.76,2.40~48.10μg/mL for Quercetin,kaempferol,isorhamnetin,respectively;The average recovery rate of quercetin,kaempferol,isorhamnetin was 96.40%,96.95%,93.48%,with the RSDs being 3.5%,3.7% and 3.7%,respectively.The average content of quercetin,kaempferol and isorhamnetin in the flowers was 0.576 1,0.265 0 and 0.173 3mg/g,respectively;The average content of quercetin,kaempferol and isorhamnetin in the leaves was 1.502 9,1.345 8 and 0.450 4mg/g,respectively;The average content of quercetin in the stems was 0.128 0mg/g but little was detectable of kaempferol and isorhamnetin in the stems.None of these 3 compounds were detected in the roots.Conclusion The method we established here was simple,stable and reproducible,which was suitable for the determination of flavonoids in Caragana altaica Kom.Pojark.