| Project/Area Number |
16590036
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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 | Nihon Pharmaceutical University |
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Principal Investigator |
YOSHIMURA Yoshihiro Nihon Pharmaceutical University, Faculty of Pharmacy, Professor, 薬学部, 教授 (00147894)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAZAWA Hiroyuki Hoshi University, Faculty of Pharmacy, Professor, 薬学部, 教授 (50150173)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | life-style diseases / diagnosis / allantoin / mass spectrometry / electron spin resonance / uric acid / 親水性相互作用クロマトグラフィー / 電気化学検出器 / スピントラッピング / 分離分析 |
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| Research Abstract |
A sensitive and selective analytical method for uric acid in saliva was developed by high-performance liquid chromatography with ultraviolet detector (HPLC-UV). The results showed that the detection limit and the quatificatiion limit (S/N = 10) of UA (S/N = 3) was 3 nM and 0.1μM, respectively. Concentrations of uric acid in the saliva of seven volunteers were 134.5±103.3μM
Allantoin is a catabolic product of purines, which is formed by the enzymatic reaction of uricase with uric acid. An accurate, sensitive and selective analytical method for the determination of allantoin in human biological samples using hydrophllic interaction chromatography with mass spectrometly (HILIC/MS) was developed. The detection limit (S/N = 3) and the quatificatiion limit (S/N = 10) were 0.03 and 0.1 μM, respectively The average recoveries of allantoin with an internal standard in human serum, urine and saliva samples were 96.9% (RSD: 13.2%), 95.2% (RSD: 9,0%) and 100.3% (RSD: 11.8%), respectively. Concentrations of allantoin in the urea, saliva and serum of six volunteers were 15.3〜163 μM, 1.46〜6.7 μM and 1.18〜3.05 μM.
Reactivity of uric acid and various free radicals as the biological oxidative stress were investigated by electron spin resonance (ESR). Uric acid had strong scavenging activity for free radicals. It was found that allantoin was produced by reaction of uric acid with superoxide anion and hydroxyl radical.
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