Laboratory for the Analysis of Medicines, Department of Pharmacy, Faculty of Medicine, CIRM, University of Liege, Avenue Hippocrate 15, B36, +3, Tower 4, 4000 Liege, Belgium; Department of Analytical Chemistry and Instrumental Analysis, Faculty of Pharmacy, "Iuliu Haţieganu" University of Medicine and Pharmacy Cluj-Napoca, 4 Louis Pasteur street, 400349 Cluj-Napoca, Romania.
Department of Analytical Chemistry and Instrumental Analysis, Faculty of Pharmacy, "Iuliu Haţieganu" University of Medicine and Pharmacy Cluj-Napoca, 4 Louis Pasteur street, 400349 Cluj-Napoca, Romania.
J Chromatogr A. 2019 Apr 12;1590:80-87. doi: 10.1016/j.chroma.2018.12.068. Epub 2019 Jan 10.
D-amino acids (AA) analysis is becoming more and more relevant for metabolomics, therefore new analytical tools need to be developed. A common approach to achieve AA enantioseparation is chiral derivatization. Among the chiral derivatization reagents, (+) or (-)-1-(9-fluorenyl) ethyl chloroformate ((+) or (-)-FLEC) has proved to be one of the most versatile. Suitable chiral selectivity for FLEC derivatives of amino acids could be obtained in reversed-phase HPLC using nonpolar stationary phases (C4, C8 and C18) and tetrahydrofuran (THF) based mobile phases. This study is meant to provide alternatives to the use of THF as organic modifier by evaluating the selectivity obtained on two phenyl based stationary phases for 19 FLEC-DL-AA pairs of diastereomers using UHPLC-MS. Several mobile phases consisting of ammonium acetate and different common organic solvents (acetonitrile (ACN), methanol (MeOH), 2-propanol (IPA)) were tested using gradient elution. Experimental design was employed for the optimization of the separation conditions. In the optimized conditions, complete chiral separation can be achieved for 18 out of 19 FLEC-DL-AAs in less than 30 min.
D-氨基酸（AA）分析在代谢组学中变得越来越重要，因此需要开发新的分析工具。实现 AA 对映体分离的一种常见方法是手性衍生化。在众多手性衍生化试剂中，(+)或(-)-1-（9-芴基）乙基氯甲酸酯（(+)或(-)-FLEC）已被证明是最通用的试剂之一。使用非极性固定相（C4、C8 和 C18）和基于四氢呋喃（THF）的流动相在反相 HPLC 中可以获得 FLEC 衍生氨基酸的合适手性选择性。本研究旨在通过使用 UHPLC-MS 评估两种苯基固定相在 19 对 FLEC-DL-AA 对映体上获得的选择性，为使用 THF 作为有机改性剂提供替代方法。使用梯度洗脱测试了由乙酸铵和几种常见有机溶剂（乙腈（ACN）、甲醇（MeOH）、2-丙醇（IPA））组成的多个流动相。实验设计用于优化分离条件。在优化条件下，19 种 FLEC-DL-AAs 中的 18 种可以在不到 30 分钟内实现完全手性分离。
