Simultaneous determination of primary and secondary D- and L-amino acids by reversed-phase high-performance liquid chromatography using pre-column derivatization with two-step labelling method
Simultaneous determination of primary and secondary D- and L-amino acids by reversed-phase high-performance liquid chromatography using pre-column derivatization with two-step labelling method
复制标题
柱前衍生两步标记法反相高效液相色谱同时测定初级和次级 D- 和 L- 氨基酸
DOI：
10.1080/09168451.2017.1340090
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Mikami Koji
中科院分区：
文献类型：
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作者：
Yokoyama Takehiko;Tokuda Masaharu;Amano Masafumi;Mikami Koji
本工作描述了一种同时测定原氨基酸和脯氨酸和脯氨酸等次级氨基酸的方法。为了消除对伯胺和仲胺同时测定的干扰，先用邻苯二醛/ n -乙酰-l-半胱氨酸（OPA/NAC）对伯胺进行衍生，然后用1-（9-芴基）氯甲酸乙酯（FLEC）对仲胺进行柱前分离衍生。这些氨基酸的荧光非对映体在室温下3分钟内获得，并在高效液相色谱柱的单次洗脱中通过改变波长同时测定。这种方法被称为“两步标记法”，对于同时测定d-和l-氨基酸是有效的。
This work describes a method for the simultaneous determination of primaryd- andl-amino acids and secondary amino acids such asd- andl-proline. In order to remove interferences in the simultaneous determination of primary and secondary amines, the primary amines were derivatized witho-phthalaldehyde/N-acetyl-l-cysteine (OPA/NAC) and subsequently with 1-(9-fluorenyl)ethyl chloroformate (FLEC) for secondary amines, in a pre-column separation derivatization technique. These fluorescent diastereomers of the amino acids were obtained within 3 min at room temperature and determined simultaneously by changing wavelengths during analysis in a single eluting run in the high-performance liquid chromatography column. This method, referred to as the “two-step labelling method,” is effective for the simultaneous determination ofd- andl-amino acids.