A rapid method for simultaneous determination of 15 flavonoids in Epimedium using pressurized liquid extraction and ultra-performance liquid chromatography
A rapid method for simultaneous determination of 15 flavonoids in Epimedium using pressurized liquid extraction and ultra-performance liquid chromatography
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DOI：
10.1016/j.jpba.2007.09.016
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发表时间：
2008-01-22
影响因子：
3.4
通讯作者：
Li, S. P.
中科院分区：
文献类型：
--
作者：
Chen, X. J.;Ji, H.;Li, S. P.
淫羊藿为小檗科淫羊藿属植物淫羊藿，为常用中药之一。黄酮类化合物被认为是其活性成分。本研究建立了超高效液相色谱法（UPLC）同时测定甘草中15种黄酮类化合物的含量，包括：山柰苷E、山柰酚-3-O-鼠李糖苷、山柰苷F、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、朝藿苷C、宝霍苷II、草霍苷C、宝霍苷VII、慈姑苷A、慈姑苷B、淫羊藿不同品种中的2“-O-鼠李糖淫羊藿苷II和宝活苷I。在沃茨Acquity UPLC BEH C18柱（50 mm x 2.1 mm I.D.，1.7 50 mM乙酸水溶液-乙腈梯度洗脱，在12 min内，各校正曲线在检测范围内线性良好（R-2 > 0.9997）。柱上检出限和定量限分别低于0.13和0.52 ng。15种分析物在3个水平下的日内和日间相对标准偏差均小于5.0%，回收率为95.0- 103.7%。该方法已成功地应用于不同品种淫羊藿中15种黄酮类化合物的定量分析。结果表明，所测黄酮类化合物的含量差异较大。对15个化合物的超高效液相色谱峰特征进行聚类分析，结果表明，37个样品可分为3大类，这与样品中黄酮类化合物的含量相一致。生成高含量簇的模拟平均色谱图以比较来自不同种和/或位置的淫羊藿样品。以淫羊藿甙A、B、C和淫羊藿苷4种黄酮类化合物为指标，对淫羊藿属药用植物进行质量控制。(C)2007 Elsevier B. V.保留所有权利。
Herba Epimedii (family Berberidaceae), Yinyanghuo in Chinese, is one of commonly used Chinese medicines. Flavonoids are considered as its active components. In this study, a rapid ultra-performance liquid chromatography (UPLC) method was developed for simultaneous determination of 15 flavonoids, including hexandraside E, kaempferol-3-O-rhamnoside, hexandraside F, epimedin A, epimedin B, epimedin C, icariin, epimedoside C, baohuoside II, caohuoside C, baohuoside VII, sagittatoside A, sagittatoside B, 2 ''-O-rhamnosyl icariside II and baohuoside I in different species of Epimedium. The analysis was performed on Waters Acquity UPLC system with an Acquity UPLC BEH C18 column (50 mm x 2.1 mm I.D., 1.7 mu m) and gradient elution of 50 mM acetic acid aqueous solution and acetonitrile within 12 min. All calibration curves showed good linearity (R-2 > 0.9997) within test ranges. The LOD and LOQ were lower than 0.13 and 0.52 ng on column, respectively. The R.S.D.s for intra- and inter-day of 15 analytes were less than 5.0% at three levels, and the recoveries were 95.0-103.7%. The validated method was successfully applied to quantitatively analyze 15 flavonoids in different species of Epimedium. The results showed there were great variations among the contents of investigated flavonoids. Hierarchical clustering analysis based on characteristics of 15 investigated compounds peaks in UPLC profiles showed that 37 samples were divided into 3 main clusters, which were in accordance with their flavonoids contents. The simulative mean chromatogram of the high content cluster was generated to compare the samples from different species and/or locations of Epimedium. Four flavonoids including epimedin A, B, C and icariin were selected as markers for quality control of the species of Epimedium used as Yinyanghuo. (C) 2007 Elsevier B.V. All rights reserved.