Petritis K, Valleix A, Elfakir C, Dreux M
Institut de Chimie Organique et Analytique, CNRS UMR 6005, Université d'Orléans, France.
J Chromatogr A. 2001 Apr 13;913(1-2):331-40. doi: 10.1016/s0021-9673(00)01268-1.
Simultaneous chiral separations of underivatized amino acids have been performed using a teicoplanin-based chiral stationary phase and ionspray tandem mass spectrometry for their ionisation and detection. Different amino acid enantiomer pairs were separated simultaneously, including those of positional isomeric amino acids (e.g., L,D-Leu/Ile, or L,D-Val/Iva). Due to the specificity of tandem mass spectrometry, co-eluting enantiomers of different amino acids could also be determined. Fifteen chiral underivatized proteinogenic and non-proteinogenic amino acids were analysed simultaneously under isocratic conditions (acetonitrile-water, 75:25) in less than 25 min. For maximum sensitivity, post-column addition of 500 mM aqueous HCOOH was necessary. Detection limits varied from 2.5 to 50 microg l(-1) depending on the amino acid. The signal vs. concentration relationship was linear for all D- and L-amino acids (0.9995 < or = r2 < or = 1) for three orders of magnitude.
使用基于替考拉宁的手性固定相和离子喷雾串联质谱进行未衍生化氨基酸的同时手性分离,以实现其电离和检测。不同的氨基酸对映体对可同时分离,包括位置异构氨基酸(如L,D-亮氨酸/异亮氨酸或L,D-缬氨酸/缬氨霉素)的对映体对。由于串联质谱的特异性,不同氨基酸的共洗脱对映体也可被测定。在等度条件下(乙腈-水,75:25),不到25分钟内可同时分析15种手性未衍生化的蛋白质原性和非蛋白质原性氨基酸。为获得最大灵敏度,柱后添加500 mM的甲酸水溶液是必要的。检测限因氨基酸而异,从2.5到50 μg l(-1)不等。所有D-和L-氨基酸的信号与浓度关系在三个数量级内呈线性(0.9995 ≤ r2 ≤ 1)。