Automated precolumn derivatization system for analyzing physiological amino acids by liquid chromatography/mass spectrometry.
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 microm Wakosil-II 3C8-100HG column (100 x 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%.
Shimbo
Kazutaka
K
Institute of Life Sciences, Ajinomoto Co., Inc., 1-1 Suzuki-cho Kawasaki-ku, Kawasaki, Japan.
Kubo
Shintaro
S
Harada
Yushi
Y
Oonuki
Takashi
T
Yokokura
Takefumi
T
Yoshida
Hiroo
H
Amao
Michiko
M
Nakamura
Mina
M
Kageyama
Naoko
N
Yamazaki
Junko
J
Ozawa
Shin-ichi
S
Hirayama
Kazuo
K
Ando
Toshihiko
T
Miura
Junkichi
J
Miyano
Hiroshi
H
D000596:Amino Acids / Q000032:analysis* / Q000097:blood / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*
D000596:Amino Acids