1. Glucose oxidase, horseradish peroxidase and phenothiazine dyes-co-adsorbed carbon felt-based amperometric flow-biosensor for glucose.
- Author
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Jiao Z, Kuang L, Komori M, Hirono M, Komuro R, Wang Y, and Hasebe Y
- Subjects
- Phenothiazines chemistry, Enzymes, Immobilized chemistry, Adsorption, Electrochemical Techniques methods, Coloring Agents chemistry, Limit of Detection, Methylene Blue chemistry, Hydrogen Peroxide chemistry, Glucose Oxidase chemistry, Glucose Oxidase metabolism, Horseradish Peroxidase chemistry, Biosensing Techniques methods, Glucose chemistry, Glucose analysis, Carbon chemistry
- Abstract
To develop an amperometric flow-biosensor for glucose, the stabilizing effect of methylene blue (MB) toward adsorbed glucose oxidase (GOx) on carbon felt (CF) was successfully applied to prepare the GOx-modified CF-based enzyme reactor combined with a horseradish peroxidase (HRP)-modified CF-based H
2 O2 detector. Upon mixing MB in the GOx-adsorption solution, the O2 -dependent GOx-activity was significantly increased with increasing concentration of MB in the GOx-adsorption solution. The GOx-immobilization protocol on CF is very straightforward [ i.e. , adsorption of the GOx/MB mixed aqueous solution for 5 min under ultrasound (US)-irradiation]. Under the optimized operational conditions ( i.e. , applied potential, 0 vs. Ag/AgCl; carrier pH, 5.0; carrier flow rate, 4.0 mL min-1 ), the resulting GOx/MB-CF-reactor and HRP/TN-CF-detector combined amperometric flow-biosensor exhibited sensitive, selective, reproducible and stable cathodic peak current responses to glucose with the following analytical performances: sensitivity, 6.22 μA mM-1 ; linear range, 0.01 to 1 mM; limit of detection, 9.6 μM (S/N = 3, noise level, 20 nA); sample throughput, 46-96 samples per h for 10-0.1 mM glucose. The developed amperometric flow-biosensor allowed the determination of glucose in beverages and liquors, and the analytical results by the sensor were in fairly good agreement with those by conventional spectrophotometry.- Published
- 2024
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