Development of Micro Analytical System of Hydroperoxides in Lipids

采用美国合成的新型试剂二苯基-1-芘基膦（DPPP），建立了生物样品中脂质氢过氧化物（HPO）的极灵敏荧光测定法。建立了一种定量测定食用油脂中总HPO的方法。试剂和样品溶液的混合物在60^ C下在黑暗中反应60分钟。反应结束后，用荧光法测定氧化DPPP。通过该方法，脂质HPO的亚纳米摩尔水平可以按样品的mg级进行测定。高效液相色谱系统采用柱后检测装置，该装置由输送试剂溶液的泵、反应线圈和荧光计组成。通过该系统，在皮摩尔水平上测定了HPO。(1)甘油三酯（TG） HPO：在ODS色谱柱上对TG- monohpo进行了分节数分离。在苯基化硅胶柱上，他们给出了一个单峰，使总tg -单hpo的测定成为可能。(2)磷脂酰胆碱(PC)-和磷脂酰乙醇胺(PE) - hpo: PC-和PE- hpo通过硅胶柱和氨基丙基柱直接连接进行分离。该系统适用于血浆样品中PC-HPO的测定。

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.