| Project/Area Number |
63860011
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Research Field |
応用生物化学・栄養化学
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| Research Institution | Tohoku University |
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Principal Investigator |
MEGURO Hiroshi Tohoku Univ., Agriculture, Professor, 農学部, 教授 (30005590)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Hiroaki Tosho, Chief Researcher, 主任研究員
AKASAKA Kazuaki Tohoku Univ., Agriculture, Associate Prof., 農学部, 助手 (10201881)
NISHIDA Yoshihiro Tohoku Univ., Agriculture, Associate Prof., 農学部, 助手 (80183896)
OHRUI Hiroshi Tohoku Univ., Agriculture, Assistant Prof., 農学部, 助教授 (20100050)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1990: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1989: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1988: ¥4,300,000 (Direct Cost: ¥4,300,000)
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| Keywords | hydroperoxide / dertermination / fluorometry / HPLC / diphenyl pyrenyl phosphine / diphenylー1-pyrenylphosphine / high performance liquid chromatography / triacyl glycerol / ジフェニルー1ーピレニルホスフィン / ヒドロペルオキシド / 蛍光分析システム |
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| Research Abstract |
Extremely sensitive fluorometry of lipid hydroperoxides (HPO) in biological samples were developed using diphenyl-1-pyrenylphosphine (DPPP), a novel reagent synthesized us.
1. A batch method is developed to determine total HPO in edible fats and oils. The mixture of the reagent and sample solutions was reacted at 60^ﾟC for 60 min in the dark. After the reaction, DPPP oxide was determined by fluorometry. By the method, subnano mole level of lipid HPO could be determined with mg order of a sample.
2. HPLC systems are developed by using post column detection unit which composed of a pump to send the reagent solution, reaction coil and a fluorometer. By the systems, HPO were determined at pico mole level.
(1) Triacylglycerol (TG) HPO : TG-monoHPO were separated to several peaks on their partition numbers on an ODS colomn. On a phenylated silicagel column, they gave a single peak to made possible to determined total TG-monoHPO.
(2) Phosphatidylcholine (PC)- and Phosphatidylethanolamine (PE) -HPO : PC- and PE-HPO were separated by two columns, a silicagel and an aminopropyl columns, connected directly. This system was applicable to determine PC-HPO in plasma samples.
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