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基于酶抑制的生物传感器中使用固定在丝网印刷电极上的灰盖鬼伞过氧化物酶对硫化物进行安培检测。

Amperometric sulfide detection using Coprinus cinereus peroxidase immobilized on screen printed electrode in an enzyme inhibition based biosensor.

机构信息

Department of Chemical and Petroleum Engineering, Sharif University of Technology, P.O. Box 11155-9465, Azadi Ave., Tehran, Iran.

Department of Chemical and Petroleum Engineering, Sharif University of Technology, P.O. Box 11155-9465, Azadi Ave., Tehran, Iran.

出版信息

Biosens Bioelectron. 2012 May 15;35(1):297-301. doi: 10.1016/j.bios.2012.03.004. Epub 2012 Mar 10.

DOI:10.1016/j.bios.2012.03.004
PMID:22472527
Abstract

In the present work, an amperometric inhibition biosensor for the determination of sulfide has been fabricated by immobilizing Coprinus cinereus peroxidase (CIP) on the surface of screen printed electrode (SPE). Chitosan/acrylamide was applied for immobilization of peroxidase on the working electrode. The amperometric measurement was performed at an applied potential of -150 mV versus Ag/AgCl with a scan rate of 100 mV in the presence of hydroquinone as electron mediator and 0.1M phosphate buffer solution of pH 6.5. The variables influencing the performance of sensor including the amount of substrate, mediator concentration and electrolyte pH were optimized. The determination of sulfide can be achieved in a linear range of 1.09-16.3 μM with a detection limit of 0.3 μM. Developed sensor showed quicker response to sulfide compared to the previous developed sulfide biosensors. Common anions and cations in environmental water did not interfere with sulfide detection by the developed biosensor. Cyanide interference on the enzyme inhibition caused 43.25% error in the calibration assay which is less than the amounts reported by previous studies. Because of high sensitivity and the low-cost of SPE, this inhibition biosensor can be successfully used for analysis of environmental water samples.

摘要

在本工作中,通过将毛栓菌过氧化物酶(CIP)固定在丝网印刷电极(SPE)表面,制备了用于测定硫化物的安培抑制生物传感器。壳聚糖/丙烯酰胺用于在工作电极上固定过氧化物酶。在存在氢醌作为电子介体和 pH 6.5 的 0.1M 磷酸盐缓冲溶液的情况下,在 150 mV 相对于 Ag/AgCl 的施加电位下进行安培测量,扫描速率为 100 mV。优化了影响传感器性能的变量,包括底物的量、介体浓度和电解质 pH。硫化物的测定可在 1.09-16.3 μM 的线性范围内实现,检测限为 0.3 μM。与以前开发的硫化物生物传感器相比,开发的传感器对硫化物的响应更快。环境水中的常见阴离子和阳离子不会干扰开发的生物传感器对硫化物的检测。氰化物对酶抑制的干扰在标定试验中导致 43.25%的误差,这小于以前研究报告的数量。由于 SPE 的高灵敏度和低成本,这种抑制生物传感器可成功用于环境水样的分析。

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