Abstract

A simple mono-enzyme biosensor has been developed for specific determination of paraoxon (POX). In this biosensor, choline oxidase (ChOx) was introduced and used as a new recognition element. Besides, by application of Prussian blue (PB), as an electron mediator, the necessity of applying oxygen or hydrogen peroxide sensor, as the internal transducer, was abated. The method was based on PB chemically modified screen-printed electrode coupled with ChOx for detection of POX as inhibitor. The concentration of H2O2 produced by ChOx was electrochemically determined by the PB modified electrode at -50 mV versus the internal screen-printed Ag pseudo-reference electrode. The decrease in current caused by the addition of inhibitor was used for evaluation of POX concentration. The experimental parameters such as the quantity of enzyme, substrate concentration and incubation time were optimized for ChOx inhibition. At the incubation time of 5 min, the biosensor response was linear from 0.1 to 1 mu M of POX with a detection limit of 0.1 mu M. (c) 2009 Elsevier B.V. All rights reserved.