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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 [mgr]m 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.
|Authors:||Shimbo, Kazutaka; Kubo, Shintaro; Harada, Yushi; Oonuki, Takashi; Yokokura, Takefumi; Yoshida, Hiroo; Amao, Michiko; Nakamura, Mina; Kageyama, Naoko; Yamazaki, Junko; Ozawa, Shin‐ichi; Hirayama, Kazuo; Ando, Toshihiko; Miura, Junkichi; Miyano, Hiroshi|
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