SiO_2/Nafion杂化膜固定酶的过氧化氢生物传感器  

Amperometric Biosensor for Hydrogen Peroxide Based on Horseradish Peroxidase Incorporated into SiO_2/Nafion Hybrid Film

作　　者：孙康[1] 王知彩[2] 李仲辉[1] 

机构地区：[1]四川教育学院化学系,四川成都611130 [2]安徽工业大学化学与化工学院,安徽马鞍山243002

出　　处：《分析测试学报》2010年第11期1185-1189,共5页Journal of Instrumental Analysis

基　　金：四川省科技计划资助项目(2008GZ0041)

摘　　要：通过溶胶-凝胶技术制备SiO2/Nafion杂化膜并固定辣根过氧化物酶,以杂化膜中Nafion固定的亚甲基蓝为辣根过氧化物酶和玻碳电极间的电子传递介体,制成了电流型过氧化氢生物传感器。探讨了杂化膜的制备条件、工作电位、pH值、温度、干扰物质等对生物传感器的影响。该生物传感器的线性响应范围为1.0×10-6～1.6×10-4mol/L,检出限(S/N=3)为6.0×10-7mol/L,达到95%稳态响应电流用时少于15s。固定化酶对过氧化氢催化反应的米氏常数为1.129 mmol/L。A highly sensitive amperometric biosensor for H2O2 was constructed by immobilizing horseradish peroxidase（HRP） into the hybrid film of sol - gel-derived SiO2 and Nation. The methylene blue（MB） immobilized in the hybrid film acted as an electron transfer mediator between horseradish peroxidase and glass carbon electrode. The preparation conditions for the hybrid membrane and the effects of applied potential, solution pH, temperature and interferents on the biosensors were discussed in details. The fabricated biosensor showed a wide linear response to hydrogen peroxide in the range of 1.0 × 10^-6 - 1.6×10^-4 mol/L with a detection limit（S/N = 3 ） of 6.0 ×10^-7 mol/L. The biosensor could reach 95% of steady-state current within 15 s and retain 93.2% of its initial activity after 2 months storage in 0. 1 mol/L pH 7.0 phosphate buffer. The Michaelis - Menten constant of the immobilized enzyme was calculated to be 1. 129 mmol/L for H2O2, which indicated that HRP incorporated into SiO2/Nafion hybrid film maintained excellent enzymatic activity and had high affinity to H2O2.

关 键 词：过氧化氢 生物传感器 溶胶-凝胶技术 SiO2/Nafion杂化膜 亚甲基蓝 

分 类 号：O657.1[理学—分析化学] TQ123.6[理学—化学]