Enantioselective determination of citrulline and ornithine in the urine of D-amino acid oxidase deficient mice using a two-dimensional high-performance liquid chromatographic system

Reiko Koga, Yurika Miyoshi, Yu Sato, Masashi Mita, Ryuichi Konno, Wolfgang Lindner, Kenji Hamase

研究成果: ジャーナルへの寄稿記事

13 引用 (Scopus)

抄録

Two-dimensional high-performance liquid chromatographic (2D-HPLC) and 2D-HPLC-mass spectrometric (2D-HPLC–MS) systems have been designed and developed for the determination of the citrulline (Cit) and ornithine (Orn) enantiomers. Several D-amino acids have already been identified as novel physiologically active molecules and biomarkers, and the enantioselective evaluation of the amounts, distributions and metabolisms of non-proteinogenic amino acids gain as well increasing interest. In the present study, highly selective analytical methods were developed using a capillary monolithic ODS column (0.53 mm i.d. x 1000 mm) for the reversed-phase separation of the target analytes from the matrix compounds in the first dimension, and a narrowbore-Pirkle type enantioselective column, KSAACSP–105 S (1.5 mm i.d. x 250 mm), was used for the enantiomer separation in the second dimension. The amino acids were analyzed after pre-column derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) and detected by the fluorescence detector and MS. The systems were applied to the urine of D-amino acid oxidase (DAO) deficient B6DAO mice and control C57BL mice to evaluate the presence and metabolism of the Cit and Orn enantiomers in mammals. As a result, all of the 4 target enantiomers (D-Cit, L-Cit, D-Orn, L-Orn) were found in the urine of both strains. The %D value of Cit (D-Cit/Cit × 100) increased about 3-fold in the urine of the DAO deficient mice and that of Orn also tended to increase with the DAO deficiency. These results were definitely confirmed by a 2D-HPLC–MS detection system. Further investigations about the biological significance of these D-isomers are currently ongoing.

元の言語英語
ページ(範囲)312-317
ページ数6
ジャーナルJournal of Chromatography A
1467
DOI
出版物ステータス出版済み - 10 7 2016

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D-Amino-Acid Oxidase
Citrulline
Ornithine
Enantiomers
Urine
Liquids
High Pressure Liquid Chromatography
Amino Acids
Metabolism
Mammals
Biomarkers
Inbred C57BL Mouse
Phase separation
Isomers
Fluorescence
Detectors
Molecules

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

これを引用

Enantioselective determination of citrulline and ornithine in the urine of D-amino acid oxidase deficient mice using a two-dimensional high-performance liquid chromatographic system. / Koga, Reiko; Miyoshi, Yurika; Sato, Yu; Mita, Masashi; Konno, Ryuichi; Lindner, Wolfgang; Hamase, Kenji.

:: Journal of Chromatography A, 巻 1467, 07.10.2016, p. 312-317.

研究成果: ジャーナルへの寄稿記事

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abstract = "Two-dimensional high-performance liquid chromatographic (2D-HPLC) and 2D-HPLC-mass spectrometric (2D-HPLC–MS) systems have been designed and developed for the determination of the citrulline (Cit) and ornithine (Orn) enantiomers. Several D-amino acids have already been identified as novel physiologically active molecules and biomarkers, and the enantioselective evaluation of the amounts, distributions and metabolisms of non-proteinogenic amino acids gain as well increasing interest. In the present study, highly selective analytical methods were developed using a capillary monolithic ODS column (0.53 mm i.d. x 1000 mm) for the reversed-phase separation of the target analytes from the matrix compounds in the first dimension, and a narrowbore-Pirkle type enantioselective column, KSAACSP–105 S (1.5 mm i.d. x 250 mm), was used for the enantiomer separation in the second dimension. The amino acids were analyzed after pre-column derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) and detected by the fluorescence detector and MS. The systems were applied to the urine of D-amino acid oxidase (DAO) deficient B6DAO− mice and control C57BL mice to evaluate the presence and metabolism of the Cit and Orn enantiomers in mammals. As a result, all of the 4 target enantiomers (D-Cit, L-Cit, D-Orn, L-Orn) were found in the urine of both strains. The {\%}D value of Cit (D-Cit/Cit × 100) increased about 3-fold in the urine of the DAO deficient mice and that of Orn also tended to increase with the DAO deficiency. These results were definitely confirmed by a 2D-HPLC–MS detection system. Further investigations about the biological significance of these D-isomers are currently ongoing.",
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AU - Miyoshi, Yurika

AU - Sato, Yu

AU - Mita, Masashi

AU - Konno, Ryuichi

AU - Lindner, Wolfgang

AU - Hamase, Kenji

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