| Project/Area Number |
58470052
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
工業分析化学
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| Research Institution | Kyoto University |
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Principal Investigator |
KOJIMA Tsugio Kyoto University, Faculty of Engineering, Professor, 工学部, 教授 (20025810)
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| Co-Investigator(Kenkyū-buntansha) |
MORISHITA Fujio Kyoto University, Faculty of Engineering, lnstnuctor, 工学部, 助手 (30026281)
ICHISE Mitsunojo Kyoto University, Faculty of Engineering, Assocate Professor, 工学部, 助教授 (00025917)
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| Project Period (FY) |
1983 – 1985
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| Project Status |
Completed (Fiscal Year 1985)
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| Budget Amount *help |
¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 1985: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1984: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1983: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Enzyme-immobilized open-tubular reactors / flow injection analysis / simultaneous determination / urate / triglycerides / トリグリセリド / グルコース |
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| Research Abstract |
Combination of advantages in our FIA method, in which enzyme-immobilized open tubular reactors are used, with one of the features of enzymatic reactions that a common product is often formed from various different enzymatic reactions leads to development of an FIA method for simultaneous determination of many species. Thus, an FIA system for that purpose consists of a single injector, a single detector and plural reactors in parallel. Since such a system is also effective for measurement of signal change before and after a reaction, an FIA system for the urate determination is constructed at first, in which a uricase immobilized reactor and a bypass tube are placed in parallel and decrease in absorbance at 290 nm is measured from two peaks appearing successively. Urate in sera in the concentration range of clinical significance could be determined with satisfactory reproducibilities. Secondly, the simultaneous determination of three species was attempted with 3-channel FIA system. Band broadening within open tubular reactor can be described by the Taylor-Aris equation corrected for the secondary flow occuring in the section of a coiled reactor. An optimal geometry of FIA system was designed by taking into consideration the resolution of the peak pairs in addition to band broadening. Lactate-, glucose- and glycerol-dehydrogenase-catalyzed reactions were chosen. The corresponding substrates were simultaneously determined by monitoring the resulting common product, NADH, fluorometrically (ex. 340 nm, em. >460 nm). Last, trigycerides and <beta>-D-glucose were determined simultaneously with an FIA system, in which all the reactions including hydrolysis of triglycerides were carried out in the stream of the carrier solution.
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