| Project/Area Number |
10672029
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Physical pharmacy
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| Research Institution | Hoshi University |
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Principal Investigator |
YOSHIMURA Yoshihiro Hoshi University, Faculty of Pharmacy, Associate Professor, 薬学部, 講師 (00147894)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Takaho Hoshi University, Faculty of Pharmacy, Research assistant, 薬学部, 助手 (80210912)
NAKAZAWA Hiroyuki Hoshi University, Faculty of Pharmacy, Professor, 薬学部, 教授 (50150173)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2000: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1999: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Nitric Oxide / Separation analysis / Electron chemical detection / Spin tapping / Electron spin resonance / ECD / フリーラジカル / 生体試料 |
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| Research Abstract |
Electron spin resonance (ESR) method was examined for determination of nitric oxide (NO) using Carboxy-PTIO.the Proposed method was 100 nM as detection limit (S/N=3) of NO, and more high sensitivety and saecificitv than these of Grease method and fluorescence method. We developed the HPLC/ECD/ESR system, which be able to separate for the many free radicals, determining each radical within 10 min by using the reverse-phase HPLC.Scince Carboxy-PTIO is easy to oxidize by reductants such as ascorbic acid, tocopherol and uric acid in the organism, good determination for NO could not obtained. Therefore, Liposome- PTIO which was prepared bv using the lecithin, examined stability for determination of NO in the biological systems. It is elucidate that Liposome-PTIO could be protected from many reductants by barrier of lipid bilayer. The good linearity is obtained between 10μM to 100μM, and detection limit (S/N=3) is 5 μM by the ECD detector and 50 nM by ESR.We applied proposed method to determination of NO of lacrimal fluid in rat suffered from conjunctivitis. As the results, generation of 0.4 μM for NO was recognized in the lacrimal fluid of rat. Formation of NO was confirmed in the aorta thoracica of the rat which caused the blood vessel relaxation.
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