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Development of automatic semi-micro simultaneus determination system for lipid hydroperoxides in human plasma

Research Project

Project/Area Number 10556028
Research Category

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

Allocation TypeSingle-year Grants
Section展開研究
Research Field 食品科学・栄養科学
Research InstitutionTOHOKU UNIVERSITY

Principal Investigator

AKASAKA Kazuaki  TOHOKU UNIVERSITY, Graduate School of Agriculturalscience, Assistant professor, 大学院・農学研究科, 助教授 (10201881)

Co-Investigator(Kenkyū-buntansha) TAKAHASHI Hiroaki  Tosoh corporation, scientific and diagnostic Technical researchist insturmails and systems, 科学計測事業部, 技術研究員
OHRUI Hiroshi  TOHOKU UNIVERSITY, Graduate school of Agriculturalscience, professor, 大学院・農学研究科, 教授 (20100050)
高橋 裕明  東ソー株式会社, 科学計測事業部, 技術研究員
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥8,700,000 (Direct Cost: ¥8,700,000)
Fiscal Year 2000: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1999: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1998: ¥4,300,000 (Direct Cost: ¥4,300,000)
Keywordslipid hydroperoxide / phosphine / fluorescence determination / High-perfomance liquid chromatography / 過酸化脂質の分析 / 高速液体クロマトグラフィー / ジフェニル-1-ピレニルホスフィン / 過酸化脂質の一斉分析 / セミミクロ液体クロマトグラフィー / ジフェニル・1・ピレニルホスフィン
Research Abstract

HPLC-post column systems for simultaneous determination of lipid hydroperoxides was developed. Lipid hydroperoxides were separated on an ODS column by using a gradient elution mode, then they reacted with diphenyl-1-pyrenylphosphine(DPPP), a fluorescence reagent for lipid hydroperoxides, in a stainless steel coil at 80 C and fluorescence intensity was monitored. By the system, eight kinds of lipid hydroperoxides were separated each other and determined at pmol levels with good reproducibility. The system was successfully down sized to semi-micro system using an 2.0mm i.d. ODS column. The system made it possible to save the amount of a reagent and the solvents by needing less than 1/10 and 1/3, respectively, of the amounts for the conventional size system without reducing sensitivity and reproducibility. By using ph-silica column instead of an ODS column, it was possible to separate lipid hydroperoxides on their class levels. These systems allowed us to inject 90 ul of sample in MeOH/chloroform(1 : 1)by slightly modification of an injection port.
By using these systems, we showed that the X(active oxygen species)-Y(catalyst)-Z(oxygen acceptor)chemiluminescence system played as a scavenging system of lipid hydroperoxides in the presence of Mn^<2+>. We also showed that some beverages had strong lipid hydroperoxide scavenging activity.

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] 赤坂和昭,大類洋: "過酸化脂質のFIAによる分析"日本油化学会誌. 10. 1053-1059 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kazuaki Akasa,Hritaka Ohta,YoshimiHanGda,HeroshiOhrui.: "Simultaneous Determination of Lipid Hydroperoxides by HPLC-post Column Systems"Biosci.Biotechnol.Biochem.,. 64. 1842-1846 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kazuaki Akasa,Hiroshi Ohrui: "Development of phosphine reagents for the high-perforance liquid chromatography-fluorometric determination of lipid hydroperoxides"Journal of Chromatography A. 881. 159-170 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kazuaki Akasa, Hiroshi Ohrui: "Flow Injection Analysis of Lipid Hydoroperoxides"NIHON YUKAGAKUKAI. 47, 10. 1053-1059 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kazuaki Asaka, Hritaka Ohta, YoshimiHanada, HiroshiOhrui: "Simultaneous Determination of Lipid Hydroperoxides by HPLC-post Column Systems"Biosci.Biotechnol.Biochem.. 64. 1842-1846 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kazuaki Akasa, Hiroshi Ohrui: "Development of phosphine reagents for the high-perforance liquid chromatography-fluorometric determination of lipid hydroperoxides"Journal of Chromatography A. 881. 159-170 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kazuaki Akasaka,Hiroshi Ohrui,: "Development of phosphine reagents for the high-performance liquid chromatographic-fluorometric determination of lipid hydroperoxides"Journal of Chromatography A. 881. 159-170 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] K.Akasaka,H.Ohta,Y.Hanada & H.Ohrui: "simultaneous determination of lipid hydroperoxides by HPLC-Post column Systems"Biosci. Biotechnol. Biochem. 63・8. 1506-1508 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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