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Estimation of free radical formation and deletion of life relation substances

自由基形成和生命关系物质缺失的估计

基本信息

批准号：
09672200
负责人：
NAKAZAWA Hiroyuki
金额：
$ 1.92万
依托单位：
Hoshi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672200/
关键词：
free radical life relation substance ESR spin trapping agent radical chromatography scavenger superoxide anion hydroxy radical スカベンチャー NOラジカル

项目摘要

The life relation substances are getting involved largely, in the generation and deletion of free radicals that the relations with cancer, diabetes, brain disease etc. which are pointed out and research is going on from the face of the disease and prevention. An individual activity oxygen speceise need to be measured well accuracy, to analyze the action system of the activity oxygens of superoxide. There are many case of activity oxygen speceis that plural free radical contributes to the reaction system that gets involved. We tried the simultaneous determination of free radicals to the separation of each frer radical by using high performance liquid chromatography (HPLC), in order to develop the accrurate analytical methods in this research. Furthermore, high sensitivity analysis became possible by using the electrochemical detector with HPLC. Also, the simultaneous an alysis of DMPO adducts of 3 kinds of free radical became possible by HPLC/ESR/ECD system that we constucted, beside the chemical generation procedure of super oxide anion, hydroxyl radical, and methyl radical by hidrogen peroxide and DMSO are established. Similarly, as a result that we used PBN as the spin trapping agents as for 3 kinds of PBN adducts of which the distinction is difficult in the ESR spectrum, the fine base line separation was achieved regarding 3 kinds of PBN addcuts by the developed system. As a result that we examied the scavenger effect of free radical by the system using the ascorbic acid, dl-α-tocopherol, β-carotin, as the antioxidant the dosage reliance nature is admitted and analyzed the scavenger reaction with the characteristic of each antioxidant. Also, this free radical generation system is conceivable as usefulness even for the free radical deletion activity measurement of the chenmicals, because even the chemical of the water-solubility and also hydrophobic property are able to melt.

生命关系物质越来越多地参与到自由基的产生和清除中，从疾病的角度和预防角度指出了自由基与癌症、糖尿病、脑病等疾病的关系，并进行了研究。为了分析超氧化物中活性氧的作用体系，需要对单个活性氧进行准确的测量。在许多情况下，复数自由基对参与的反应体系有贡献。为了在本研究中建立准确的分析方法，我们尝试用高效液相色谱法（HPLC）同时测定自由基以分离各自由基。此外，利用高效液相色谱电化学检测器进行高灵敏度分析成为可能。此外，通过构建的HPLC/ESR/ECD体系，建立了过氧化氢和DMSO生成超氧阴离子、羟基自由基和甲基自由基的化学生成过程，实现了对三种自由基的DMPO加合物的同时分析。同样，由于我们使用PBN作为自旋捕获剂，对于ESR光谱中难以区分的3种PBN加合物，所开发的系统对3种PBN加合物实现了精细的基线分离。因此，我们以抗坏血酸、dl-α-生育酚、β-胡萝卜素为抗氧化剂，考察了该体系对自由基的清除作用，承认了其剂量依赖性，并根据各抗氧化剂的特性分析了其清除作用。此外，这种自由基生成系统甚至可以用于化学物质的自由基删除活性测量，因为即使化学物质的水溶性和疏水性也能够熔化。