Šimek Petr, Hušek Petr, Zahradníčková Helena
Laboratory of Analytical Biochemistry and Metabolomics, Biology Centre, Czech Academy of Sciences, České Budějovice, Czech Republic.
Methods Mol Biol. 2019;2030:237-251. doi: 10.1007/978-1-4939-9639-1_18.
Gas chromatography (GC) is a commonly used technique in amino acid analysis (AAA). However, one of the requirements of the application of GC for AAA is a need for the polar analytes to be converted into their volatile, thermally stable derivatives. In the last two decades, alkyl chloroformates (RCFs) have become attractive derivatization reagents. The reagents react immediately with most amino acid functional groups in aqueous matrices, and the process can easily be coupled with liquid-liquid extraction of the resulting less polar derivatives into immiscible organic phase. Here we describe a simple protocol for in situ derivatization of amino acids with heptafluorobutyl chloroformate (HFBCF) followed by subsequent chiral as well as nonchiral GC/MS (mass spectrometric) analysis on a respective nonpolar fused silica and an enantioselective Chirasil-Val capillary column.
气相色谱法（GC）是氨基酸分析（AAA）中常用的技术。然而，将GC应用于AAA的要求之一是需要将极性分析物转化为其挥发性、热稳定的衍生物。在过去二十年中，氯甲酸烷基酯（RCF）已成为有吸引力的衍生化试剂。这些试剂能立即与水性基质中的大多数氨基酸官能团发生反应，并且该过程可轻松与将所得极性较小的衍生物液 - 液萃取到不混溶的有机相中相结合。在此，我们描述了一种用七氟丁基氯甲酸酯（HFBCF）对氨基酸进行原位衍生化的简单方案，随后分别在非极性熔融石英柱和对映选择性Chirasil-Val毛细管柱上进行手性以及非手性GC/MS（质谱）分析。
