Development of Micro Analytical System of Hydroperoxides in Lipids

脂质中氢过氧化物微量分析系统的研制

• 批准号: 63860011
• 负责人: MEGURO Hiroshi
• 金额: $ 4.48万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B).
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63860011/
• 关键词: 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

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.

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。建立了批量法测定食用油脂中总HPO的方法。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，在皮科水平上测定了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。

项目成果

Hiroshi MEGURO: "Development of new fluorometrical reagents applicable to food and biological materials" Proceedings of the Third Korea-Japan Joint Symposium on Analytical Chemistry. 153-158 (1989)

Hiroshi MEGURO：“开发适用于食品和生物材料的新型荧光试剂”第三届韩日分析化学联合研讨会论文集。

Kazuaki AKASAKA: "Fluorometric Determination of Triacylglycerol Hydroperoxides at Picomole Levels by HPLC PostーColumn Method with Diphenylー1ーpyrenylphosphine" J.Chromatogr.

Kazuaki AKASAKA：“采用二苯基-1-芘基膦的 HPLC 柱后方法荧光测定皮摩尔水平的三酰甘油氢过氧化物”J.Chromatogr。

Kazuaki AKASAKA: "An aromatic phosphine reagent for the hplc-fluorescence determination of hydroperoxides -determination of phosphatidylcholine hydroperoxides in human plasma -" Analytical Letters. 21 (6). 965-975 (1988)

Kazuaki AKASAKA：“用于氢过氧化物的 hplc 荧光测定的芳香膦试剂 - 人血浆中磷脂酰胆碱氢过氧化物的测定 -” 分析快报。

Kazuaki AKASAKA: "AN AROMATIC PHOSPHINE REAGENT FOR THE HPLCーFLUORESCENCE DETERMINATION OF HYDROPEROXIDES ー DETERMINATION OF PHOSPHATIDYLCHOLINE HYDROPEROXIDES IN HUMAN PLASMA ー" ANALYTICAL LETTERS. 21. 965-975 (1988)

Kazuaki AKASAKA：“用于氢过氧化物的 HPLC 荧光测定的芳香族膦试剂 - 人血浆中磷脂酰胆碱氢过氧化物的测定 -”分析快报 21. 965-975 (1988)。

Kazuaki AKASAKA: "A Simple Fluoromety of Hydroperoxides in Oils and Foods" Agric. Biol. Chem.

Kazuaki AKASAKA：“油和食品中氢过氧化物的简单荧光测定”农业。