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Highly sensitive chemiluminescence FIA of the minor components in relation to diabetes involving a bioreactor

Research Project

Project/Area Number 08672642
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory medicine
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

TABATA Masayoshi  Kyoto University College of Medical Technology Assistant Professor, 医療技術短期大学部, 助教授 (70115880)

Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1997: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1996: ¥1,500,000 (Direct Cost: ¥1,500,000)
Keywords3-hydroxybutyrate / NADH oxidase / 3-hydroxbutyrate dehydrogenase / 1,5-AG / pyranose oxidase / glucokinase / chemiluminescence / coimmobilized enzyme column / 固定化酵素 / グルコキナーゼ / バイオリアクター / FIA / NADHアキシダーゼ
Research Abstract

Increasing the sensitivity for chemiluminescence, and a chemiluminescence-FIA of serum 3-hydroxybutyrate (3-HBA) and 1,5-anhydroglucitol (1,5-AG) using a bioreactor were tried. The detection sensitivity for H_2O_2 was increased up to about 0.1 mu mol/mu 1 of sample by the introduction of a non-pulse flow pumps, an improvement of a flow cell, and intercepting the light which goes into the detecktor along the tubes from the outside For the determination of serum 3-HBA,the coimmobilized 3-hydroxybutyrate dehydrogenase (3-HBADH)/NADH oxidase (NOX) column with a weight ratio 5 : 1 was the most efficient for determining serum 3-HBA.The column hydrolyzed all the 1.5 mM 3-HBA,and showed linearity of the data up to 1.5 mM 3-HBA.Forever, the chemiluminescence intensity obtained by the column was about twice as much as those by the sequential and the mixed-bed 3-HBADH/NOX columns, because 3-HBADH and NOX in the coimmobilized 3-HBADH/NOX column were in such close proximity that both progresses proceeded quickly.
Pyranose oxidase (PROD) oxidized not only glucose but also 1,5-AG to produce H_2O_2. The oxidation rate for 1,5-AG was far slower than that for glucose, but because the column contained a large quantity of PROD,the oxidation rate for 1,5-AG increased to 70% that for glucose. When the coimmobilized glucokinase (GK)/pyruvate kinase (PK) column was used for the removal of serum glucose, GK phosphorylated not only glucose but also 1,5-AG.But, the phosphorylation rate for 1,5-AG was far lower compared with that for glucose. The most efficient column for 1,5-AG was the coimmobilized GK/PK column with a weight ratio GK 1000U : PK 0.28ml. The column removed 99.99% glucose in a sample of 1 mu 1 injected. The results from serum 1,5-AG correlated satisfactorily with those by other method.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Masayoshi Tabata: "Removal of glucose in serum by a bioreactor and its application to the determination of serum 1,5-anhydroglucitol" Proceedings of the X VI International Congress of Clinical Chemistry. 376- (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 田畑 勝好: "固定化酵素カラムを装着したフローインジェクション分析のLDH反応解析への応用" 臨床病理. 44(Suppl.). 75- (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 田畑 勝好: "バイオリアクターと化学発光法を装着したFIAによる血清1,5-AGの高感度分析" 臨床化学. 26(Supp.3). 77- (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 田畑 勝好: "バイオリアクターと化学発光法を組込んだFIAによる血清グルコースの高感度分析" 日本臨床検査自動化学会会誌. 22(4). 413- (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 田畑 勝好: "検査試薬としての酵素の使い方" 化学と生物. 36(1). 51-56 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 田畑 勝好: "老人病(老化)と臨床検査" 臨床病理. 45(3). 251-253 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 田畑 勝好: "新編臨床検査講座18 臨床化学" 医歯薬出版株式会社, (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 田畑 勝好: "天然高分子の新展開(分担執筆:診断用材料)" シ-エムシ-, (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Tabata: "Removal of glucose in serum by a bioreactor and its application to the determination of serum 1,5-anhydroglucitol" Proceedings of the X VI international Congress of Clinical Chemistry. 376 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Tabata: "The application of flow injection analysis involving an immobilized enzyme column to the analysis of LDH reaction" Jpn.J.Clin.Pathol.44(Suppl.). 75 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Tabata: "Highly sensitive flow injection analysis of serum 1,5-anhydroglucitol involving a bioreactor and chemiluminescence" Jpn.J.Clin.Chem.26(Suppl.3). 77 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Tabata: "Highly sensitive FIA of serum glucose using a bioreactor and chemiluminescence" Jpn.J.Clin.Lab.Automa.22(4). 413 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Tabata: "How to use enzyme as examination reagents" KAKAKU TO SEIBUTSU. 36. 51-56 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Tabata: "Geriatric diseases (aging) and clinical pathology" Jpn.J.Clin.Pathol.45. 251-253 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 田畑勝好: "バイオリアクターと化学発光法を装着したFIAによる血清1,5-AGの高感度分析" 臨床化学. 26(Supp.3). 77 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 田畑勝好: "バイオリアクターと化学発光法を組込んだFIAによる血清グルコースの高感度分析" 日本臨床検査自動化学会会誌. 22(4). 413 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 田畑勝好: "検査試薬としての酵素の使い方" 化学と生物. 36(1). 51-56 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 田畑勝好: "老人病(老化)と臨床検査" 臨床病理. 45(3). 251-253 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 田畑勝好: "天然高分子の新展開(分担執筆:診断用材料)" シ-エムシ-, (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nasayoshi Tabata: "Removal of glucose in serum by a bioreactor and its application to the determination of serum 1,5-anhydroglucitol" Proceedings of the X VI International Congress of Clinical Chemistry. 376 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 田畑勝好: "固定化酵素カラムを装着したフローインジェクション分析のLDH反応解析への応用" 臨床病理. 44(Suppl.). 75 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 田畑勝好: "新編臨床検査講座18 臨床化学" 医歯薬出版株式会社, (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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