Determination of d- and l-enantiomers of threonine and allo-threonine in mammals using two-step high-performance liquid chromatography

Hua Zhao, Kenji Hamase, Akiko Morikawa, Zongyin Qiu, Kiyoshi Zaitsu

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23 Citations (Scopus)

Abstract

A sensitive and selective method for the determination of four threonine (Thr) isomers (l-Thr, d-Thr, l-allo-Thr and d-allo-Thr) in mammalian tissues has been established using two-step high-performance liquid chromatography. This method includes the precolumn fluorescence derivatization of amino acids with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), and the separation using a combination of a reversed-phase column and a chiral column. The calibration ranges of d-Thr, d-allo-Thr and l-allo-Thr spiked in the rat cerebellum sample are 2.5 fmol-5 pmol per injection, and that of l-Thr is 50 fmol-50 pmol. Within-day and day-to-day precisions of the determination of the four Thr isomers are approximately 5% in the rat cerebellum. By using this method, the tissue distributions of d-Thr, d-allo-Thr and l-allo-Thr in mammals have been demonstrated for the first time in rats, and found that significant amounts of d-Thr and d-allo-Thr are present in the frontal brain areas and urine. Among the 12 tissues tested, the highest amounts of d-Thr (0.85 ± 0.05 nmol/g wet tissue) and d-allo-Thr (5.01 ± 0.32 nmol/g wet tissue) were found in the corpus striatum. l-allo-Thr was not present in any of the tested tissues and physiological fluids.

Original languageEnglish
Pages (from-to)245-250
Number of pages6
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume810
Issue number2
DOIs
Publication statusPublished - Oct 25 2004

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

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