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D-アミノ酸テーラーメイド計測プレートの開発

Research Project

Project/Area Number 19659008
Research Category

Grant-in-Aid for Exploratory Research

Allocation TypeSingle-year Grants
Research Field Physical pharmacy
Research InstitutionKyushu University

Principal Investigator

濱瀬 健司  Kyushu University, 大学院・薬学研究院, 准教授 (10284522)

Project Period (FY) 2007 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2008: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2007: ¥1,800,000 (Direct Cost: ¥1,800,000)
Keywords薬学 / 分析科学 / 生体分子 / タンパク質
Research Abstract

平成19年度に試作したD-アスパラギン酸、D-ロイシン迅速計測プレートについて、平成20年度では再現性、定量性を評価し、分析法としてのバリデートを行った。アミノ酸標準品による検討では、D-アスパラギン酸計測プレートにおける検量線は1mL当たり0.016から16μmolの範囲で良好な相関を示し、ウェル間における相対標準偏差は4.8%、プレート間における相対標準偏差は4.5%であった。ラット血中D-アスパラギン酸定量についてHPLC法との相関を検討した結果、回帰直線の傾きは1.09、Y切片は0.44であり、ほぼY=Xの直線となった。一方、D-ロイシン計測プレートにおける検量線は血清中濃度にして0.1mMから10mMの範囲で良好な定量性が認められた。ウェル間における相対標準偏差は13.2%であった。両プレートとも、L-アミノ酸による定量妨害は殆ど受けない事が示された。これらのELISAプレートを利用し、ラット血中におけるD-アスパラギン酸とD-ロイシンの動態解析を行った。各D-アミノ酸は頚静脈より投与し、経時的に血中濃度推移を検討した。その結果、D-アスパラギン酸、D-ロイシン共に投与後一分で最高濃度となった後経時的に減少し、D-アスパラギン酸では30分、D-ロイシンは60分で約3分の1に減衰した。これらの定量値はHPLC法による定量値と良く一致しており、本研究において開発したD-アミノ酸計測ELISAプレートは、実試料に適用可能な優れた迅速計測法として今後の利用が期待される。

Report

(2 results)
  • 2008 Annual Research Report
  • 2007 Annual Research Report
  • Research Products

    (16 results)

All 2009 2008 2007

All Journal Article (6 results) (of which Peer Reviewed: 6 results) Presentation (9 results) Book (1 results)

  • [Journal Article] Enantioseleotive visualization of D-alanine in rat anterior pituitary gland : localization to ACTH-secreting cells2009

    • Author(s)
      Sachise ETOH, Kenji HAMASE, et al.
    • Journal Title

      Analytical and Bioanalytical Chemistry 393

      Pages: 217-223

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Simple and rapid genotyping of D-amino acid oxdase gene recognizing a crucial variant in the ddY strain using microchip electrophoresis2009

    • Author(s)
      Yosuke TOJO, Kenji HAMASE, et al.
    • Journal Title

      Journal of Separation Science 32

      Pages: 430-436

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Circadian changes of D-alanine and related compounds in rats and the effect of restricted feeding on their amounts2008

    • Author(s)
      Akiko MORIKAWA, Kenji HAMASE, et al.
    • Journal Title

      Journal of Chromatography B 875

      Pages: 168-173

    • NAID

      10028008387

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Automated and simultaneous two-dimensional micro-high-performance liquid chromatographic determination of proline and hydroxyproline enantiomers in mammals2008

    • Author(s)
      Yosuke TOJO, Kenji HAMASE, et al.
    • Journal Title

      Journal of Chromatography B 875

      Pages: 174-179

    • NAID

      10028008429

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Circadian changes of D-alanine and related compounds in rats and theeffect of restricted feeding on their amounts2008

    • Author(s)
      Akiko MORIKAWA, Kenji HAMASE, et. al.
    • Journal Title

      Journal of Chromatography B (In press)

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Immunohistochemical localization of D-alanine to β-cells in rat pancreas2007

    • Author(s)
      Akiko MORIKAWA, Kenji HAMASE, et. al.
    • Journal Title

      Biochemical and Biophysical Research Communications 355

      Pages: 872-876

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Presentation] Comprehensive enantiomer analysis of all proteinogenic amino acids using fully automated two-dimensional micro-HPLC2008

    • Author(s)
      Kenji HAMASE, et al.
    • Organizer
      32^<nd> International Symposium on High Perform ance Liquid Phase Separations and Related Techniques
    • Place of Presentation
      Baltimore, USA
    • Related Report
      2008 Annual Research Report
  • [Presentation] Rapid enantiomer separations of all proteinogenic amino acids and their comprehensive 2D-HPLC determination in biological samples2008

    • Author(s)
      Kenji HAMASE, et al.
    • Organizer
      2O^<th>International Symposium on Chirality
    • Place of Presentation
      Geneva, Switzerland
    • Related Report
      2008 Annual Research Report
  • [Presentation] 哺乳類における微量D・アミノ酸の二次元一斉分析法構築2008

    • Author(s)
      浜瀬健司, 他
    • Organizer
      第4回D-アミノ酸研究会学術講演会
    • Place of Presentation
      名古屋
    • Related Report
      2008 Annual Research Report
  • [Presentation] Two-dimensional HPLC analysis of D-amino acids in mammals2008

    • Author(s)
      Kenji HAMASE, et al.
    • Organizer
      33^<rd> International Symposium on High Performanoe Liquid Phase Separations and Related Techniques
    • Place of Presentation
      Kyoto, Japan
    • Related Report
      2008 Annual Research Report
  • [Presentation] タンパク質構成全アミノ酸光学異性体の二次元HPLC一斉分析と哺乳類体内含量の解析2008

    • Author(s)
      浜瀬健司, 他
    • Organizer
      第81回日本生化学会大会
    • Place of Presentation
      神戸
    • Related Report
      2008 Annual Research Report
  • [Presentation] Establishment of a Comprehensive Micro-2D Analysis System for Highly Sensitive Determination of Branched Aliphatic Amino Acid Enantiomers in Human Plasma and Urine2007

    • Author(s)
      Kenji Hamase, et. al.
    • Organizer
      31^<st> International Symposium on High Performance Liquid Phase Separations and Related Techniques
    • Place of Presentation
      Ghent, Belgium
    • Related Report
      2007 Annual Research Report
  • [Presentation] Ultra-sensitive determination of multiple D-amino acids in mammals using two-dimensional micro-HPLC with fluorescence detection2007

    • Author(s)
      Kenji Hamase, et. al.
    • Organizer
      10^<th> International Congress on Amino Acids and Proteins
    • Place of Presentation
      Kallithea, Greece
    • Related Report
      2007 Annual Research Report
  • [Presentation] アミノ酸光学異性体の高感度二次元HPLC分析法開発と哺乳類体内におけるD-アミノ酸の可能性2007

    • Author(s)
      浜瀬 健司
    • Organizer
      第207回液体クロマトグラフィー研究懇談会
    • Place of Presentation
      野田市
    • Related Report
      2007 Annual Research Report
  • [Presentation] 二次元HPLCを用いる哺乳類体内の微量D-アミノ酸分析2007

    • Author(s)
      浜瀬 健司, 財津 潔
    • Organizer
      第80回日本生化学会大会
    • Place of Presentation
      横浜市
    • Related Report
      2007 Annual Research Report
  • [Book] 薬学分析科学の最前線2009

    • Author(s)
      浜瀬 健司, 他
    • Total Pages
      185
    • Publisher
      株式会社 じほう
    • Related Report
      2008 Annual Research Report

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Published: 2007-04-01   Modified: 2016-04-21  

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