Fully automated reagent peak-free liquid chromatography fluorescence analysis of highly polar carboxylic acids using a column-switching system and fluorous scavenging derivatization.
Fully automated reagent peak-free liquid chromatography fluorescence analysis of highly polar carboxylic acids using a column-switching system and fluorous scavenging derivatization.
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DOI：
10.1002/jssc.201200692
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发表时间：
2013
影响因子：
3.1
通讯作者：
K. Todoroki;Hiroki Hashimoto;Kazuyuki Machida;Miki Itoyama;Tadashi Hayama;H. Yoshida;H. Nohta;M. Nakashima;M. Yamaguchi
中科院分区：
文献类型：
--
作者：
K. Todoroki;Hiroki Hashimoto;Kazuyuki Machida;Miki Itoyama;Tadashi Hayama;H. Yoshida;H. Nohta;M. Nakashima;M. Yamaguchi
在本研究中，我们将柱开关系统与氟清除衍生化方法相结合，开发了一种全自动试剂无峰LC荧光检测方案，用于分析高极性羧酸。在此方法中，高极性羧酸在1-乙基-3-（3-二甲氨基丙基）碳二亚胺和1-羟基- 1h -苯并三唑存在下，与荧光1-芘甲胺衍生。剩余的未反应试剂用2-（全氟辛基）乙基异氰酸酯标记，然后使用放置在分析反相柱前的氟柱切换系统选择性地去除。在得到的色谱图中，氟标记的未反应试剂的信号完全不存在；因此，它不影响每种酸的定量，特别是在20分钟前洗脱的酸。所检酸的检出限（S/N = 3）在每针4.0 ~ 22 fmol之间。我们将这种方法应用于2型糖尿病模型小鼠和对照小鼠尿液样品中高极性羧酸的比较分析。
In this study, we combined a column-switching system with a fluorous scavenging derivatization method to develop a fully automated reagent peak-free LC fluorescence detection protocol for the analysis of highly polar carboxylic acids. In this method, highly polar carboxylic acids were derivatized with fluorescent 1-pyrenemethylamine in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 1-hydroxy-1H-benzotriazole. Residual excess of the unreacted reagent was tagged with 2-(perfluorooctyl)ethyl isocyanate and then removed selectively using a fluorous column-switching system placed in front of an analytical reversed-phase column. The signal of the fluorous-tagged unreacted reagent was completely absent in the resulting chromatograms; therefore, it did not interfere with the quantification of each acid especially those eluted before 20 min. The detection limits (S/N = 3) for the examined acids were in the range from 4.0 to 22 fmol per injection. We have applied this method to comparative analysis of highly polar carboxylic acids in urine samples obtained from diabetes mellitus type-II model mice and their control.