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研制了一种新型流动注射化学发光(CL)双酶传感器,用于葡萄糖的检测.该传感器将掺杂金纳米粒子(GNPs)的壳聚糖膜包覆在硅烷化试剂预处理的玻璃微珠上,用于吸附固定葡萄糖氧化酶(GOD)和辣根过氧化物酶(HRP).葡萄糖在GOD的催化下发生氧化反应生成H2O2,生成的H2O2在HRP的催化作用下与鲁米诺发生反应,并产生化学发光信号.实验表明,壳聚糖中掺杂的GNPs不仅能够有效的吸附酶分子并保持其生物活性,还对Luminol-H2O2-HRP化学发光体系具有增敏作用.通过化学发光光谱和紫外光谱表征,详细研究了固定化GNPs增强Luminol-H2O2-HRP体系的化学发光机理.在优化的实验条件下,该传感器对葡萄糖检测的线性范围为0.01～6.0mmol/L,检测限为5.0μmol/L(3σ).将所建立的方法用于临床血清样品中葡萄糖含量的测定,获得了满意的结果.
A novel flow injection chemiluminescence (CL) bi-enzyme sensor was developed for the detection of glucose.The sensor encapsulated chitosan films coated with gold nanoparticles (GNPs) in silanization reagent-pretreated glass microbeads (GOD) and horseradish peroxidase (HRP). Glucose oxidized by GOD to form H2O2, and the resulting H2O2 reacts with luminol under the action of HRP , And generate chemiluminescence signals.Experiments show that GNPs doped in chitosan not only can effectively adsorb enzyme molecules and maintain its biological activity, but also sensitize Luminol-H2O2-HRP chemiluminescence system. And UV spectroscopy, the chemiluminescence mechanism of Luminol-H2O2-HRP system was enhanced by immobilized GNPs in detail.The linear range of this sensor for glucose detection was 0.01-6.0 mmol / L and the detection limit was 5.0 under the optimized experimental conditions μmol / L (3σ) .The proposed method was applied to the determination of glucose in clinical serum samples, and satisfactory results were obtained.