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Automated precolumn derivatization system for analyzing physiological amino acids by liquid chromatography/mass spectrometry
Author(s) -
Shimbo Kazutaka,
Kubo Shintaro,
Harada Yushi,
Oonuki Takashi,
Yokokura Takefumi,
Yoshida Hiroo,
Amao Michiko,
Nakamura Mina,
Kageyama Naoko,
Yamazaki Junko,
Ozawa Shinichi,
Hirayama Kazuo,
Ando Toshihiko,
Miura Junkichi,
Miyano Hiroshi
Publication year - 2010
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1346
Subject(s) - chemistry , derivatization , chromatography , mass spectrometry , electrospray , high performance liquid chromatography , amino acid , biochemistry
An automated method for high‐throughput amino acid analysis, using precolumn derivatization high‐performance liquid chromatography/electrospray mass spectrometry (HPLC/ESI‐MS), was developed and evaluated. The precolumn derivatization step was performed in the reaction port of a home‐built auto‐sampler system. Amino acids were derivatized with 3‐aminopyridyl‐ N ‐hydroxysuccinimidyl carbamate, and a 3 μm Wakosil‐II 3C8‐100HG column (100 × 2.1 mm i.d.) was used for separation. To achieve a 13 min cycle for each sample, the derivatization and separation steps were performed in parallel. The results of the method evaluation, including the linearity, and the intra‐ and inter‐precision, were sufficient to measure physiological amino acids in human plasma samples. The relative standard deviations of typical amino acids in actual human plasma samples were below 10%. Copyright © 2009 John Wiley & Sons, Ltd.