Development of Micro Analytical System of Hydroperoxides in Lipids

采用自行合成的新试剂二苯基-1-芘基膦（DPPP），建立了生物样品中脂质过氧化氢（HPO）的高灵敏荧光分析法。建立了食用油脂中总过氧化氢的批量测定方法。将试剂和样品溶液的混合物在黑暗中在60 ° C下反应60分钟。反应后，通过荧光法测定DPPP氧化物。该方法可测定亚纳摩尔级的脂质过氧化氢，样品量为毫克级. HPLC系统是通过使用柱后检测单元开发的，柱后检测单元由用于发送试剂溶液的泵、反应线圈和荧光计组成。利用该系统，在皮摩尔水平上测定了HPO。(1)三酰甘油（TG）HPO：TG-单HPO在ODS柱上按其分配数分离成多个峰。在苯基化硅胶柱上，它们产生一个单峰，从而可以测定总TG-单HPO。(2)磷脂酰胆碱（PC）-和磷脂酰乙醇胺（PE）-HPO：PC-和PE-HPO通过硅胶柱和氨丙基柱直接连接的两个柱分离。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.