Disposable biosensors for determination of biogenic amines
Disposable biosensors for determination of biogenic amines
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DOI：
10.1016/j.aca.2010.03.012
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发表时间：
2010-04-14
影响因子：
6.2
通讯作者：
Julia Arcos-Martinez, M.
中科院分区：
文献类型：
--
作者：
Asuncion Alonso-Lomillo, M.;Dominguez-Renedo, Olga;Julia Arcos-Martinez, M.
本工作报道了基于单胺氧化酶(MAO)/辣根过氧化物酶（HRP）和二胺氧化酶(DAO)/辣根过氧化物酶（HRP）的生物传感器，采用丝网印刷碳电极测定生物胺（BA）。这些酶被共价固定在碳工作电极上，之前用羟基丁二酰亚胺和碳二酰亚胺修饰芳基重氮盐。检测是通过测量中间体二茂羟甲基铁在低电位下（250mv vs丝网印刷Ag/AgCl参比电极）还原的阴极电流进行的。采用实验设计方法对酶固定化的实验条件和影响计时电流的主要变量进行了优化。在这些最佳条件下，对一次性生物传感器进行了表征。以DAO/HRP和MAO/HRP为基础的生物传感器，组胺的线性响应范围分别为0.2 ~ 1.6 μ M和0.4 ~ 2.4 μ M。基于DAO/HRP和MAO/HRP的生物传感器的灵敏度相对标准偏差分别为10%和11%，重复性高。MAO/HRP生物传感器的检测能力分别为0.18 +/- 0.01 μ M和0.40 +/- 0.04 μ M (alpha = 0.05和beta = 0.005)，并分析了MAO/HRP生物传感器对不同BA的灵敏度。最后，将所研制的生物传感器应用于鱼类样品中总胺含量的测定。(C) 2010 Elsevier B.V.版权所有
This work reports monoamine oxidase (MAO)/horseradish peroxidase (HRP) and diamine oxidase (DAO)/horseradish peroxidase (HRP) based biosensors using screen-printed carbon electrodes for the determination of biogenic amines (BA). The enzymes have been covalently immobilized onto the carbon working electrode, previously modified by an aryl diazonium salt, using hydroxysuccinimide and carbodiimide. The detection has been performed by measuring the cathodic current due to the reduction of the mediator hydroxymethylferrocene at a low potential, 250 mV vs screen-printed Ag/AgCl reference electrode. The experimental conditions for the enzymes immobilization, as well as for the main variables that can influence the chronoamperometric current have been optimized by the experimental design methodology. Under these optimum conditions, the disposable biosensors have been characterized. A linear response range from 0.2 up to 1.6 mu M and from 0.4 to 2.4 mu M of histamine was obtained for DAO/HRP and MAO/HRP based biosensors, respectively. The biosensor construction was highly reproducible, yielding relative standard deviations of 10% and 11% in terms of sensitivity for DAO/HRP and MAO/HRP based biosensors, respectively. The capability of detection, 0.18 +/- 0.01 mu M in the case of DAO/HRP and 0.40 +/- 0.04 mu M (alpha = 0.05 and beta = 0.005) for MAO/HRP based biosensors, and the biosensor sensitivity towards different BA has also been analyzed. Finally, the developed biosensors have been applied to the determination of the total amine content in fish samples. (C) 2010 Elsevier B.V. All rights reserved.