Selective and sensitive liquid chromatographic determination method of 5-hydroxyindoles with fluorous and fluorogenic derivatization
Selective and sensitive liquid chromatographic determination method of 5-hydroxyindoles with fluorous and fluorogenic derivatization
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氟荧光衍生化选择性灵敏液相色谱测定5-羟基吲哚的方法
DOI：
10.1016/j.jpba.2015.06.003
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发表时间：
2015
影响因子：
3.4
通讯作者：
Hitoshi Nohta
中科院分区：
文献类型：
--
作者：
Yohei Sakaguchi;Jun Ikenaga;Hideyuki Yoshida;Tadashi Hayama;Miki Itoyama;Kenichiro Todoroki;Osamu Imakyure;Masatoshi Yamaguchi;Hitoshi Nohta
建立了同时测定5-羟基吲哚(5-His；5-羟色氨酸、5-羟色胺、N-乙酰基-5-羟色胺、5-羟基吲哚-3-乙酸和5-羟色醇)的高效液相色谱(LC)方法。该方法包括用4-(3′，3′，4′，4′，5′，5′，6′，6′，7′，7′，8′，8′，9′，9′，10′，10′，10′-heptadecafluorodecyl)benzylamine(HFBA)柱前衍生化和使用氟液色谱柱分离衍生物。在本研究中，通过简单的衍生化过程得到了稳定的5-His与HFBA的苯并恶唑衍生物；它们的荧光性质使高灵敏的检测成为可能。此外，只有5-组氨酸的HFBA衍生物通过氟相互作用被选择性地保留在含氟液晶柱上，全氟烷基化合物之间显示出亲和力，而非氟化合物则没有。在30分钟内分离，20μL进样量中5-His的检出限为1.2~14 Fmol(S/N=0.3)。此外，将该方法应用于健康人血浆中5-His的分析。
A liquid chromatographic (LC) method with improved selectivity for the simultaneous determination of 5-hydroxyindoles (5-HIs; 5-hydroxytryptophan, 5-hydroxytryptamine,N-acetyl-5-hydroxytryptamine, 5-hydroxyindole-3-acetic acid, and 5-hydroxytryptophol) is described. This method involves precolumn derivatization with 4-(3′,3′,4′,4′,5′,5′,6′,6′,7′,7′,8′,8′,9′,9′,10′,10′,10′-heptadecafluorodecyl)benzylamine (HFBA) and separation of the derivatives using a fluorous LC column. In this study, stable benzoxazole derivatives of 5-HIs with HFBA have been obtained by a simple derivatization procedure; their fluorescent properties enabled highly sensitive detection. In addition, only the HFBA derivatives of 5-HIs has been selectively retained on the fluorous LC column via fluorous interaction whereby perfluoroalkyl compounds show affinities with each other, while the non-fluorous compounds did not. The HFBA derivatives were separated within 30 min and the detection limits for 5-HIs in a 20-μL injection volume were 1.2–14 fmol (S/N= 3). Furthermore, this method was applied to the analysis of 5-HIs in the human plasma from healthy subjects.