Department of Life Science and Green Chemistry, Graduate School of Engineering, Saitama Institute of Technology, 1690 Fukaya, Saitama 369-0293, Japan.
Department of Life Science and Green Chemistry, Faculty of Engineering, Saitama Institute of Technology, 1690 Fukaya, Saitama 369-0293, Japan.
Anal Methods. 2024 Aug 29;16(34):5883-5895. doi: 10.1039/d4ay01028k.
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 HO detector. Upon mixing MB in the GOx-adsorption solution, the O-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 [, adsorption of the GOx/MB mixed aqueous solution for 5 min under ultrasound (US)-irradiation]. Under the optimized operational conditions (, applied potential, 0 Ag/AgCl; carrier pH, 5.0; carrier flow rate, 4.0 mL min), 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; 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.
为了开发葡萄糖的安培流动生物传感器，成功地应用亚甲蓝 (MB) 对碳纤维毡 (CF) 上吸附的葡萄糖氧化酶 (GOx) 的稳定作用，以制备结合辣根过氧化物酶 (HRP) 修饰的 CF 基 HO 检测器的 GOx 修饰 CF 基酶反应器。在 GOx 吸附溶液中混合 MB 时，随着 GOx 吸附溶液中 MB 浓度的增加，O 依赖性 GOx 活性显著增加。在 CF 上进行 GOx 固定化的方案非常简单 [，在超声 (US) 照射下吸附 GOx/MB 混合水溶液 5 分钟]。在优化的操作条件下（，施加电位为 0 Ag/AgCl；载体 pH 值为 5.0；载体流速为 4.0 mL min），所得的 GOx/MB-CF 反应器和 HRP/TN-CF 检测器组合安培流动生物传感器对葡萄糖表现出敏感、选择性、重现性和稳定的阴极峰电流响应，具有以下分析性能：灵敏度为 6.22 μA mM；线性范围为 0.01 至 1 mM；检测限为 9.6 μM（信噪比为 3，噪声水平为 20 nA）；对于 10-0.1 mM 葡萄糖，样品通量为每小时 46-96 个样品。所开发的安培流动生物传感器可用于测定饮料和酒类中的葡萄糖，并且传感器的分析结果与传统分光光度法相当吻合。
