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

油脂产品和食物中的脂质不断受到有氧氧和光的破坏。这导致质量劣化并引起安全问题。因此，抑制这种氧化和评估氧化损伤是至关重要的。本研究的目的是开发一种方法，通过该方法可以测量油脂和脂质的氧化程度，具有高特异性，灵敏度，速度和简单性。它还旨在开发和评估可以有效抑制这种氧化的新型抗氧化剂。脂质过氧化产物从脂肪酸，磷脂酰胆碱和胆固醇酯定量区域，立体和光学特异性与高效液相色谱法（HPLC）配备紫外和化学发光检测器。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确定参与脂质过氧化的活性氧化剂和机制。此外，分析胆固醇酯的不同类别的脂肪酸。采用二苯基芘基膦（DPPP）建立了一种超灵敏的分析方法。通过DPPP和脂质过氧化物的反应按化学计量形成的DPPP氧化物被分析并应用于脂质过氧化物的定量。通过这种方法证实，脂质过氧化物在细胞中的位置可以用共聚焦荧光显微镜可视化。该方法具有良好的应用前景和发展前景。各种化合物的抗氧化能力进行了评估，从脂质过氧化物通过上述方法测量和潜在的新型抗氧化剂正在开发中。

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.