Development of Micro Analytical System of Hydroperoxides in Lipids

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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.