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Development and Application of Quantitative Evaluation System for Oxidative Damage of Lipids

脂质氧化损伤定量评价系统的开发与应用

基本信息

批准号：
08555220
负责人：
NIKI Etsuo
金额：
$ 6.98万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

项目摘要

Lipids in oil products and foods are continuously damaged by aerobic oxygen and light. This results in quality deterioration and causes safety problems. Consequently, to inhibit such oxidation and to evaluate oxidative damage are of prime importance. The objective of this research is to develop a method by which the extent of oxidation of oils and lipids can be measured with high specificity, sensitivity, speed, and simplicity. It has been also aimed at developing and evaluating novel antioxidants which can effectively inhibit such oxidation. Lipid peroxidation products from fatty acids, phosphatidylcholines and cholesteryl esters were quantified regio-, stereo- and optical-specifically with a high performance liquid chromatography (HPLC) equipped with UV and chemiluminescence detector. The quantitative analyses have been successfully applied to oils, micelles, liposomes, lipoproteins, plasma and cells. This enabled us to distinguish and identify the active oxidants and mechanisms involved in lipid peroxidations. Furthermore, cholesterol esters were analyzed for different classes of fatty acids. Diphenylpyrenylphosphine (DPPP) was used to develop a ultra-sensitive analytical method. DPPP oxide formed stoichiometrically by the reaction of DPPP and lipid peroxides was analyzed and applied to the quantification of lipid peroxides. It was confirmed by this method that the location of lipid peroxides in the cells could be visualized with a conforcal fluorescence microscope. This method could be quite promising for further application and development. The antioxidant capacities of various compounds were evaluated from the lipid peroxides measured by the above methods and a potential novel antioxidant is now being developed.

油品和食物中的脂质不断受到有氧和光照的破坏。这会导致质量下降，并造成安全问题。因此，抑制这种氧化并评估氧化损伤是最重要的。这项研究的目的是开发一种方法，用来测量油脂的氧化程度，具有高特异性、敏感性、快速和简便性。它还旨在开发和评估能够有效抑制这种氧化的新型抗氧化剂。脂肪酸、磷脂酰胆碱和胆固醇酯的脂质过氧化产物用配备UV和化学发光检测器的高效液相色谱(HPLC)进行区域、立体和光学特异性定量。定量分析已成功地应用于油、胶束、脂质体、脂蛋白、血浆和细胞。这使我们能够区分和鉴定参与脂质过氧化的活性氧化剂和机制。此外，还对不同种类的脂肪酸进行了胆固醇酯分析。利用二苯基吡喃(DPPP)建立了一种超灵敏分析方法。分析了DPPP与过氧化脂质反应生成化学计量的DPPP氧化物，并将其应用于过氧化脂质的定量。用这种方法证实，用强制荧光显微镜可以观察到细胞内过氧化脂质的位置。该方法具有广阔的应用前景和发展前景。用上述方法测定的过氧化脂质评价了各种化合物的抗氧化能力，目前正在开发一种潜在的新型抗氧化剂。