Simultaneous determination of hydrophilic amino acid enantiomers in mammalian tissues and physiological fluids applying a fully automated micro-two-dimensional high-performance liquid chromatographic concept

Kenji Hamase, Yurika Miyoshi, Kyoko Ueno, Hai Han, Junzo Hirano, Akiko Morikawa, Masashi Mita, Tsuneaki Kaneko, Wolfgang Lindner, Kiyoshi Zaitsu

研究成果: ジャーナルへの寄稿学術誌査読

97 被引用数 (Scopus)

抄録

A validated two-dimensional HPLC system combining a microbore-monolithic ODS column and a narrowbore-enantioselective column has been established for a sensitive and simultaneous analysis of hydrophilic amino acid enantiomers (His, Asn, Ser, Gln, Arg, Asp, allo-Thr, Glu and Thr) and the non-chiral amino acid, Gly, in biological samples. To accomplish this goal, the amino acids were first tagged with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) to the respective fluorescent NBD derivatives which were separated in the first dimension by a micro-reversed-phase column. The automatically collected fractions of the target peaks were then transferred to the second dimension consisting of a Pirkle type enantioselective column generating separation factors higher than 1.13 for all the enantiomeric target analytes. The system was validated using standard amino acids and a rat plasma sample, and analytically satisfactory calibration and precision results were obtained. The present 2D-HPLC system enables the fully automated determination of hydrophilic amino acid enantiomers in mammalian samples. The d-isomers of all the investigated 9 amino acids were found in rat urine but at various enantiomeric ratios.

本文言語英語
ページ(範囲)1056-1062
ページ数7
ジャーナルJournal of Chromatography A
1217
7
DOI
出版ステータス出版済み - 2月 12 2010

!!!All Science Journal Classification (ASJC) codes

  • 分析化学
  • 生化学
  • 有機化学

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