刘洋+钱丽华

摘 要：目的：建立HPLC法测定三叶青中槲皮素和山奈酚的方法。方法：三叶青组培物样品经甲醇超声提取，HLB固相萃取小柱净化，在色谱条件：色谱柱Varian ODS C18（4.6mm×250mm×5μm）、柱温30℃、流动相0.2%磷酸水-甲醇、流速1.0mL/min、检测波长360nm下进行测定。结果：槲皮素和山奈酚的方法检出限和线性范围分别为0.05mg/kg和0.05～50μg/mL，且线性范围内的相关系数均大于0.999 9，加标回收率为71.9%～105.8%，相对标准偏差为1.5%～3.4%。结论：HPLC法结合固相萃取技术可用于测定三叶青中槲皮素和山奈酚的含量，方法检出限低，结果准确可靠，重复性好。

关键词：HPLC；固相萃取；三叶青；槲皮素；山奈酚

中图分类号 R284 文献标识码 A 文章编号 1007-7731（2015）17-26-04

Study on the Determination of Quercetin and Kaempferol in Radix tetrastigmae by High Performance Liquid Chromatography

Liu Yang et al.

（Hangzhou Academy of Agricultural Sciences，Hangzhou 310024，China）

Abstract：Objective：A high performance liquid chromatography method for determination of quercetin and kaempferol in Radix tetrastigmae was studied. Method：The samples of tissue cultures were extracted with methanol by ultrasonic and purified with HLB solid phase extraction. Then，the analysis was performed on a Varian ODS C18（4.6mm×250mm×5μm）chromatographic column with methanol and 0.2% phosphoric acid as mobile phase at a flow-rate of 1.0mL/min，the column temperature was 30℃ and the detection wavelength was 360nm.Results：method detection limits for quercetin and kaempferol were both 0.05mg/kg，and good linearity for them over the range of 0.05～50μg/mL（r>0.999 9）. The average recovery and RSD of the method for quercetin and kaempferol were 71.9%～105.8% and 1.5%～3.4%，respectively. Conclusion：high performance liquid chromatography（HPLC）method combined with solid phase extraction technology can be used for the determination of quercetin and kaempferol in Radix tetrastigmae with low method detection limits，high accuracy and good repeatability.

Key words：High performance liquid chromatography；Solid phase extraction；Radix tetrastigmae；Quercetin；Kaempferol

三叶青（Tetrastigma hemsleyanum Diels et Gilg）是葡萄科崖爬藤属多年生蔓生藤本植物，为中国特有珍稀中药植物，是一种民间常用的中草药，其性凉味苦，具有清热解毒、祛风化痰、活血止痛的功效[1-2]。化学分析研究发现，三叶青中主要含有槲皮素、山奈酚、崖爬藤甙崖、异崖爬藤甙等黄酮类化合物，同时含有氨基酸类、萜类、强心苷类、甾体类、多糖类、有机酸类、挥发油、油脂以及丰富的微量元素，药理学研究表明，三叶青黄酮类提取物具有抗肿瘤细胞增殖和诱导细胞凋亡等的作用[3-4]。目前，鉴于三叶青葫芦块根人工栽培技术瓶颈尚未突破以及野生资源过度采挖造成濒危、被列入了浙江省首批农作物种质资源保护名录品种，利用生物技术来生产三叶青中抗癌黄酮等次生代谢物不失为一条很好的途径。因此，这就需要明确三叶青组培物中黄酮类化合物的含量，为三叶青的质量控制提供理论依据。……