| Project/Area Number |
05640686
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
分離・精製・検出法
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| Research Institution | The University of Tokushima |
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Principal Investigator |
MOTONAKA Junko The University of Tokushima Faculty of Engineering Associate Professor, 工学部, 助教授 (40035811)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1993: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Cholesterol / cholesterol ester / Micro-enzyme sensors |
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| Research Abstract |
Micro-enzyme sensors using cholesterol oxidase (EC 1.1.3.6) and cholesterol esterase (EC 3.1.1.13) were developed for measuring cholesterol and cholesterol ester. The platinum micro-sensors (platinum diameter, 50 mum) were etched in hot aqua regia to create a cavity at their tip. A porous composite material prepared from acetylene black and Teflon emulsion was packed into this cavity and the redox mediator [Os (bpy)_3] (PF_6)_2 was monitored by cyclic voltammetry in the potential range of 200 - 900 mV.The micro-sensors were dipped overnight in buffer solution containing the desired enzyme to immobilize it on the tip by adsorption. Calibration curves for measurements of cholesterol and cholesterol ester, the effects of pH,temperature, and concomitant compounds, the life-time of the micro-sensors, and their availability for measuring cholesterol and cholesterol ester in urine were examined. Under optimal conditons, the response of the sensors were linear in concentration ranges of 5 muM - 0.47 mM cholesterol and 2 muM - 1.00 mM cholesterol ester.
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