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Studies on Bio-active Large Molecules by Multi-stage Detection Chromatography

Research Project

Project/Area Number 02453060
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 工業分析化学
Research InstitutionNagoya University

Principal Investigator

HARAGUCHI Hiroki  Nagoya Univ. Fac. of Engineering, Professor, 工学部, 教授 (70114618)

Co-Investigator(Kenkyū-buntansha) HIROSE Akio  Nagoya Univ., Fac. of Engineering Research Associate, 工学部, 助手 (30109300)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1991: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1990: ¥5,200,000 (Direct Cost: ¥5,200,000)
KeywordsMulti-stage detection chromatography / Trace metals / Metalloenzymes / Chlorophile compounds / Humic acid-like substance / ICP-AES / Simultaneous multielement analysis / Natural waters / ICPーAES / 生体高分子 / 生理活性 / ICP発光分析法 / フォトダイオ-ドアレイ検出器 / ミオグロビン
Research Abstract

A combined analytical system for separation of bio-active large molecules in natural water by high performance liquid chromatography with detection by UV absorption, fluorescence and ICP-AES (inductively coupled plasma atomic emission spectrometry) has been developed. In ICP-AES detection the time-sequence measurement with 40 elements-simultaneous detection was explored, which allowed to measure multielement detection chromatograms. Metalloenzymes such as alkaline phosphatase (zinc enzyme), carboxypeptidase (zinc enzyme), catalase (iron enzyme), chlorophile compounds (magnesium-contained), and humic acid-like substances in pond water were of interest in the present research. In the case of metalloenzymes pond water was concentrated by ca. 10-1000 times as the analysis sample. Furthermore an on-line enzymatic activity measurement system was developed, where metalloenzymes were separated by size-exclusion chromatography and specific enzymatic activity was measured in the in-situ reaction column using some substrates. By this system 10^<-9>-10^<-12> g metalloenzymes in natural waters were determined successfully. As for chlorophile compounds (Chl), Chl-a, Chl-b, Chl-c and their derivatives were determined by fluorescence and ICP-AES detections after methanol extraction/reverse-phase chromatographic separation. In the study on metal-containing large molecules in natural water, the molecules with molecular weight larger than 10,000 were concentrated by 1000-1500 times with ultrafiltration, and separated by size exclusion chromatography. As the results, three kinds of humic acid-like substances were detected and it was found that some specific metal ions were combined in each molecular constituent. Further study is, however, required in order to elucidate the chemical forms of those large molecules.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] Akio Hirose: "Analysis of Myoblobin by Microcolumn Chromatofocusing with the Aid of Size Exclusion Chromatography and Photodiode Array Detection" Journal of High-Resolution Chromatography. 14. 420-423 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Wenzhi Hu: "Analysis of Nuclotides by High-Performance Liquid Chromatography with Phosphorus-Selective Detection" Journal of Chromatography. 557. 441-449 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Yukihiro Esaka: "Separation Characteristics of Whale Myoglobin by Chromatofocusing and Size Exclusion Chromatography with Aid of a Photodiode Array Detector" Analytical Sciences. 7. 205-208 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Akio Hirose: "On-line HPLC Determination of Enzymatic Activity of Alkaline Phosphatase Using Spectrofluorometric Detection" Chemistry Letters.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Reiji Ihara: "Flow Injection Analysis of Carboxypeptidase A Activity in Natural Water with Fluorometric Detection" Chemistry Letters.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hiroki Haraguchi: "On-line Post-Column Fluorometric Detection of Carboxypeptidase A Activity in High Performance Liquid Chromatography" Analytica Chimica Acta.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hiroki Haraguchi: "Atomic Absorption Spectrometry" Elsevier, 529 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 原口 紘〓: "分析化学" 丸善, 281 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] AKIO Hirose: "Analysis of Myoglobin by Microcolumn Chromatofocusing with the Aid of Size Exclusion Chromatography and Photodiode Array Detection" Journal of High-Resolution Chromatography. 14. 420-423 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Wenzhi Hu: "Analysis of Nucleotides by High-Performance Liquid Chromatography with Phosphorus-Selective Detection" Journal of Chromatography. 557. 441-449 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Yukihiro Esaka: "Separation Characteristics of Whale Myoglobin by Chromatofocusing and Size Exclusion Chromatography with Aid of a Photodiode Array Detector" Analytical Sciences. 7. 205-208 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Akio Hirose: "On-line HPLC Determination of Enzymatic Activity of Alkaline Phosphatase Using Spectrofluorometric Detection" Chemistry Letters.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Reiji Ihara: "Flow Injection Analysis of Carboxypeptidase A Activity in Natural Water with Fluorometric Detection" Chemistry Letters.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hiroki Haraguchi: "On-line Post-Column Fluorometric Detection of Carboxypeptidase A Activity in High Performance Liquid Chromatography" Analytica Chimica Acta.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hiroki Haraguchi: Elsevier. "Atomic Absorption Spectrometry", 529 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hiroki Haraguchi: Maruzen. "Analytical Chemistry", 281 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Akio Hirose: "Analysis of Myoglobin by Microcloumn Chromatofocusing with the Aid of Size Exclusion Chromatography and Photodiode Array Detection" Journal of HighーResolution Chromatography. 14. 420-423 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Wenzhi Hu: "Analysis of Nucleotides by HighーPerformane Liquid Chromatography with PhosphorusーSelective Detection" Journal of Chromatography. 557. 441-449 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Yukihiro Esaka: "Separation Characteristics of Whale Myoglobin by Chromatofocusing and Size Exclustion Chromatography with Aid of a Photodiode Array Detector" Analytical Sciences(Supplement). 7. 205-208 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Akio Hirose: "Onーline HPLC Determination of Enzymatic Activity of Alkalime Phosphatase Using Spectrofluorometric Detection" Chemistry Letters.

    • Related Report
      1991 Annual Research Report
  • [Publications] Reiji Ihara: "Flow Injection Analysis of Carboxypeptidase A Activity in Natural Water with Fluorometric Detection" Chemistry Letters.

    • Related Report
      1991 Annual Research Report
  • [Publications] Hiroki Haraguchi: "Onーline PostーColumn Fluorometric Detection of Carboxypeptidase A Activity in High Performance Liquid Chromatography" Analytica Chimica Acta.

    • Related Report
      1991 Annual Research Report
  • [Publications] Hiroki Haraguchi: "Atomic Absorption Spectrometry" Elsevier, 529 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 原口 紘耋: "分析化学" 丸善, 281 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] E.Munaf: "Speciation of mercury compounds in waste water by microcolumn liquid chromatography using a preconcentration column with cold vapour atomic absorption spectrometric detection" Analytica Chimica Acta. 235. 399-404 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] E.Munaf: "Determination of total mercury concentration in wastewater by continuous microflow analysis with cold vapor atomic absorption spectrometry" The Science of the Total Environment. 99. 205-209 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] E.Munaf: "Digestion method for the determination of mercury in continuous microflow analysis" Analytical Sciences. 6. 313-314 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] A.Hirose: "Enzymatic assay of alkaline phosphatase in natural water by onーline HPLC system" Chemistry Letters.

    • Related Report
      1990 Annual Research Report
  • [Publications] Y.Esaka: "Separation and characterization of whale myoglobin by chromatofocusing/size exclusion chromatography using a photodiode array detection system" Analytica Chimica Acta.

    • Related Report
      1990 Annual Research Report
  • [Publications] H.Haraguchi: "Selective detection of chlorophyl compounds by HPLC using UVーVIS absorption,fluorescence and ICPーAES detection techniques" Chemistry Letters.

    • Related Report
      1990 Annual Research Report
  • [Publications] H.Haraguchi: "Proceedings of JSPS/NUS Joint Seminar on Analytical Chemistry" World Scientific, 349 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] H.Haraguchi: "Atomic Absorption Spectrometry" Pergamon Press,

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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