| Summary: | 碩士 === 國立中興大學 === 材料科學與工程學系所 === 98 === Determination of diglyceride (DG) level in blood is important in clinical applications. A single use, disposable iridium-nano particle contained biosensor was developed in this study. DG is first hydrolyzed by lipase, and the released glycerol is then carried out the enzymatic reaction catalyzed by glycerol kinase and glycerol 3-phosphate oxidase in sequence producing hydrogen peroxide. The hydrogen peroxide is monitored at an applied potential of +0.5 V versus the Ag/AgCl reference electrode operating in an amperometric mode. The optimal condition of enzyme activity is lipase (22U mL-1), glycerol kinase (1U mL-1) and glycerol 3-phosphate oxidase (6U mL-1), respectively. Lipase, GK were added in test medium and shaken in order to obtain a homogeneous mixture, and then allowed to incubate at 37℃ for 1 hr. Glycerol 3-phosphate oxidase is covalently immobilized on the iridium-contained carbon working electrode using immobilization of 2% glutaraldehyde solution. Electrochemical measurement shows that the disposable biosensor detects DG concentration amperometrically which respond well and has a good linear relationship between response current and DG concentration in both phosphate buffer solution and bovine serum. The addition of surfactant 0.28% (v/v) Triton X-100 can enhance the performance of this system.
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