Determination of d-serine and d-alanine in the tissues and physiological fluids of mice with various d-amino-acid oxidase activities using two-dimensional high-performance liquid chromatography with fluorescence detection

Yurika Miyoshi, Kenji Hamase, Yosuke Tojo, Masashi Mita, Ryuichi Konno, Kiyoshi Zaitsu

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Abstract

Two-dimensional-HPLC procedures have been established for the sensitive and selective determination of d-serine (d-Ser) and d-alanine (d-Ala), and their amounts in the tissues and physiological fluids of mice with various d-amino-acid oxidase (DAO) activities have been demonstrated. These two d-amino acids are modulators of the N-methyl-d-aspartate receptor mediated neurotransmission, and the alterations in their amounts following the changes in the DAO activity are matters of interest. After pre-column derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), the d-amino acids were determined by the 2D-HPLC system with fluorescence detectors. As the first dimension, a microbore-monolithic-ODS column (750 mm × 0.53 mm I.D.) was adopted and a self-packed narrowbore-Pirkle type enantioselective column (Sumichiral OA-2500S, 250 mm × 1.5 mm I.D.) was selected for the second dimension. The lower limits of quantitation of d-Ser and d-Ala were 500 amol, and the within-day and day-to-day precisions were less than 6.8%. Using these methods, the amounts of d-Ser and d-Ala in 6 brain tissues, 4 peripheral tissues, serum and urine of mice having various DAO activities were determined; the amounts of these d-amino acids were drastically increased with a lowering of the DAO activity except for the cases of d-Ser in the frontal brain regions. The present micro-2D-HPLC procedures are powerful tools for the determination of small amounts of d-Ser and d-Ala in mammalian samples, and the obtained results would be useful for developing novel drugs that modulate the DAO activity, such as DAO inhibitors, against neuronal diseases.

Original languageEnglish
Pages (from-to)2506-2512
Number of pages7
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume877
Issue number24
DOIs
Publication statusPublished - Aug 15 2009

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High performance liquid chromatography
Alanine
Serine
Oxidoreductases
Fluorescence
High Pressure Liquid Chromatography
Tissue
Amino Acids
Fluids
Brain
Synaptic Transmission
Modulators
Urine
Detectors
Serum

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

Cite this

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title = "Determination of d-serine and d-alanine in the tissues and physiological fluids of mice with various d-amino-acid oxidase activities using two-dimensional high-performance liquid chromatography with fluorescence detection",
abstract = "Two-dimensional-HPLC procedures have been established for the sensitive and selective determination of d-serine (d-Ser) and d-alanine (d-Ala), and their amounts in the tissues and physiological fluids of mice with various d-amino-acid oxidase (DAO) activities have been demonstrated. These two d-amino acids are modulators of the N-methyl-d-aspartate receptor mediated neurotransmission, and the alterations in their amounts following the changes in the DAO activity are matters of interest. After pre-column derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), the d-amino acids were determined by the 2D-HPLC system with fluorescence detectors. As the first dimension, a microbore-monolithic-ODS column (750 mm × 0.53 mm I.D.) was adopted and a self-packed narrowbore-Pirkle type enantioselective column (Sumichiral OA-2500S, 250 mm × 1.5 mm I.D.) was selected for the second dimension. The lower limits of quantitation of d-Ser and d-Ala were 500 amol, and the within-day and day-to-day precisions were less than 6.8{\%}. Using these methods, the amounts of d-Ser and d-Ala in 6 brain tissues, 4 peripheral tissues, serum and urine of mice having various DAO activities were determined; the amounts of these d-amino acids were drastically increased with a lowering of the DAO activity except for the cases of d-Ser in the frontal brain regions. The present micro-2D-HPLC procedures are powerful tools for the determination of small amounts of d-Ser and d-Ala in mammalian samples, and the obtained results would be useful for developing novel drugs that modulate the DAO activity, such as DAO inhibitors, against neuronal diseases.",
author = "Yurika Miyoshi and Kenji Hamase and Yosuke Tojo and Masashi Mita and Ryuichi Konno and Kiyoshi Zaitsu",
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journal = "Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences",
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T1 - Determination of d-serine and d-alanine in the tissues and physiological fluids of mice with various d-amino-acid oxidase activities using two-dimensional high-performance liquid chromatography with fluorescence detection

AU - Miyoshi, Yurika

AU - Hamase, Kenji

AU - Tojo, Yosuke

AU - Mita, Masashi

AU - Konno, Ryuichi

AU - Zaitsu, Kiyoshi

PY - 2009/8/15

Y1 - 2009/8/15

N2 - Two-dimensional-HPLC procedures have been established for the sensitive and selective determination of d-serine (d-Ser) and d-alanine (d-Ala), and their amounts in the tissues and physiological fluids of mice with various d-amino-acid oxidase (DAO) activities have been demonstrated. These two d-amino acids are modulators of the N-methyl-d-aspartate receptor mediated neurotransmission, and the alterations in their amounts following the changes in the DAO activity are matters of interest. After pre-column derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), the d-amino acids were determined by the 2D-HPLC system with fluorescence detectors. As the first dimension, a microbore-monolithic-ODS column (750 mm × 0.53 mm I.D.) was adopted and a self-packed narrowbore-Pirkle type enantioselective column (Sumichiral OA-2500S, 250 mm × 1.5 mm I.D.) was selected for the second dimension. The lower limits of quantitation of d-Ser and d-Ala were 500 amol, and the within-day and day-to-day precisions were less than 6.8%. Using these methods, the amounts of d-Ser and d-Ala in 6 brain tissues, 4 peripheral tissues, serum and urine of mice having various DAO activities were determined; the amounts of these d-amino acids were drastically increased with a lowering of the DAO activity except for the cases of d-Ser in the frontal brain regions. The present micro-2D-HPLC procedures are powerful tools for the determination of small amounts of d-Ser and d-Ala in mammalian samples, and the obtained results would be useful for developing novel drugs that modulate the DAO activity, such as DAO inhibitors, against neuronal diseases.

AB - Two-dimensional-HPLC procedures have been established for the sensitive and selective determination of d-serine (d-Ser) and d-alanine (d-Ala), and their amounts in the tissues and physiological fluids of mice with various d-amino-acid oxidase (DAO) activities have been demonstrated. These two d-amino acids are modulators of the N-methyl-d-aspartate receptor mediated neurotransmission, and the alterations in their amounts following the changes in the DAO activity are matters of interest. After pre-column derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), the d-amino acids were determined by the 2D-HPLC system with fluorescence detectors. As the first dimension, a microbore-monolithic-ODS column (750 mm × 0.53 mm I.D.) was adopted and a self-packed narrowbore-Pirkle type enantioselective column (Sumichiral OA-2500S, 250 mm × 1.5 mm I.D.) was selected for the second dimension. The lower limits of quantitation of d-Ser and d-Ala were 500 amol, and the within-day and day-to-day precisions were less than 6.8%. Using these methods, the amounts of d-Ser and d-Ala in 6 brain tissues, 4 peripheral tissues, serum and urine of mice having various DAO activities were determined; the amounts of these d-amino acids were drastically increased with a lowering of the DAO activity except for the cases of d-Ser in the frontal brain regions. The present micro-2D-HPLC procedures are powerful tools for the determination of small amounts of d-Ser and d-Ala in mammalian samples, and the obtained results would be useful for developing novel drugs that modulate the DAO activity, such as DAO inhibitors, against neuronal diseases.

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U2 - 10.1016/j.jchromb.2009.06.028

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JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

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