Determination of isoflavones in red clover and related species by high-performance liquid chromatography combined with ultraviolet and mass spectrometric detection
Determination of isoflavones in red clover and related species by high-performance liquid chromatography combined with ultraviolet and mass spectrometric detection
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DOI：
10.1016/j.chroma.2003.08.001
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发表时间：
2003-10-24
影响因子：
4.1
通讯作者：
Simon, JE
中科院分区：
文献类型：
--
作者：
Wu, QL;Wang, MF;Simon, JE
建立了高效液相色谱-紫外-电喷雾电离-质谱检测器（HPLC-UV-ESI-MSD）测定红三叶草（Trifolium pratense L.）及相关物种中异黄酮的方法。采用LC-MSD在MS和MS-MS模式下，通过其分子离子和特征碎片离子峰对分离的异黄酮（包括苷元、苷和丙二酸苷）进行单独分析和鉴定，并与标准异黄酮进行比较。红三叶草中共检出31种异黄酮。还首次在相关物种 T repense L.（白三叶草）、T Hybridum L.（三叶草）和 T Campestre Schreber（啤酒花三叶草）中鉴定出多种异黄酮。基于反相 HPLC，酸性水解提取物中的所有 10 种异黄酮苷元、黄豆苷元、芒柄花素、金雀异黄酮、假水牛黄素、黄豆黄素、毛蕊异黄苷、普鲁尼丁、生物鸡丁 A、伊洛酮和草黄素在 40 分钟内成功分离，并通过 UV 和 MS 检测器单独定量。对于 10 种目标化合物，UV 检测的研究浓度范围为 24 至 12500 ng/ml，MS 检测的浓度范围为 6 至 3125 ng/ml，并且每种异黄酮的标准曲线都实现了良好的线性（UV 中 r(2) > 0.999，MS 中 r(2) > 0.99）。这 10 种化合物的准确度和重复性 (n = 10) 均在 15% 以内。这是第一个报道的能够同时定量红三叶草和相关物种中所有 10 种异黄酮苷元的方法。 (C) 2003 Elsevier B.V. 保留所有权利。
High-performance liquid chromatography-UV-electrospray ionization-mass spectrometric detector (HPLC-UV-ESI-MSD) method for determination of isoflavones in red clover (Trifolium pratense L.) and related species has been developed. The separated isoflavones including aglycones, glycosides and glycoside malonates, were individually analyzed and identified by their molecular ions and characteristic fragment ion peaks using LC-MSD under MS and MS-MS mode, and in comparison with the standard isoflavones. A total of 31 isoflavones were detected in red clover. Several isoflavones were also identified for the first time in related species, T repense L. (white clover), T hybridum L. (alsike clover) and T campestre Schreber (hop trefoil). Based on reversed phase HPLC, all 10 isoflavone aglycones, daidzein, formononetin, genistein, pseudobaptigenin, glycitein, calycosin, prunetin, biochanin A, irilone and pratensein in acidic hydrolyzed extracts were successfully separated within 40 min and quantified individually by UV and MS detectors. For the 10 target compounds, the investigated concentrations ranged from similar to24 to similar to12500 ng/ml for UV detection and similar to6 to similar to3125 ng/ml for MS detection, and good linearities (r(2) > 0.999 for UV and r(2) > 0.99 for MS) for standard curves were achieved for each isoflavone. The accuracy and repeatability (n = 10) were within 15% for these 10 compounds. This is the first method reported that enables the simultaneous quantitation of all 10 isoflavone aglycones in red clover and related species. (C) 2003 Elsevier B.V. All rights reserved.