|Budget Amount *help
¥3,000,000 (Direct Cost : ¥3,000,000)
Fiscal Year 1999 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 1998 : ¥700,000 (Direct Cost : ¥700,000)
Fiscal Year 1997 : ¥1,500,000 (Direct Cost : ¥1,500,000)
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.