A Screen‐Printed, Amperometric, Biosensor Array for the Detection of Organophosphate Pesticides Based on Inhibition of Wild Type, and Mutant Acetylcholinesterases, from Drosophila melanogaster

Abstract Screen‐printed carbon electrodes (SPCEs) modified with cobalt phthalocyanine (CoPC) have been used as base transducers in the construction of amperometric pesticide biosensors. Six individual biosensors were fabricated by depositing wildtype (WT) acetylcholinesterase (AChE) from Drosophila melanogaster or one of five mutant forms (B02, B03, B04, B421, and B65) of this enzyme, onto the surfaces of CoPC‐SPCEs; these constituted the amperometric biosensor array. The enzyme converts acetylthiocholine into its electroactive product thiocholine, which is detected at only 0 V vs. Ag/AgCl at the CoPC‐SPCEs. The measurement step is performed using chronoamperometry. In the presence of an organophosphate (OP) pesticide, the enzyme is inhibited, which leads to a decrease in thiocholine production and a corresponding decrease in anodic current. This decrease is proportional to the logarithm of the pesticide concentration. Calibration studies were performed with the biosensor array using five OPs of interest to the food industry, namely omethoate, malaoxon, dichlorvos, chlorpyrifos‐methyl‐oxon, and pirimiphos‐methyl‐oxon. It was found that different inhibition patterns occurred for the five OPs indicating the possibility of identifying and quantifying these compounds in food samples. It should be added that there was no detrimental affect on the biosensor response from wheat or apple extracts.

[1]  P Taylor,et al.  Three distinct domains in the cholinesterase molecule confer selectivity for acetyl- and butyrylcholinesterase inhibitors. , 1993, Biochemistry.

[2]  John P. Hart,et al.  Recent developments in the design and application of screen-printed electrochemical sensors for biomedical, environmental and industrial analyses , 1997 .

[3]  Jean-Louis Marty,et al.  Adsorption: an easy and efficient immobilisation of acetylcholinesterase on screen-printed electrodes , 2003 .

[4]  John P. Hart,et al.  Voltammetric and amperometric studies of thiocholine at a screen-printed carbon electrode chemically modified with cobalt phthalocyanine: studies towards a pesticide sensor , 1994 .

[5]  J. Marty,et al.  Amperometric biosensors based on nafion coated screen-printed electrodes for the determination of cholinesterase inhibitors. , 2000, Talanta.

[6]  J. Marty,et al.  Improved multianalyte detection of organophosphates and carbamates with disposable multielectrode biosensors using recombinant mutants of Drosophila acetylcholinesterase and artificial neural networks. , 2000, Biosensors & bioelectronics.

[7]  Y. Boublik,et al.  Acetylcholinesterase engineering for detection of insecticide residues. , 2002, Protein engineering.

[8]  Jean-Louis Marty,et al.  Disposable cholinesterase biosensor for the detection of pesticides in water-miscible organic solvents , 2001 .

[9]  Jean Louis Marty,et al.  Acetylcholine enzyme sensor for determining methamidophos insecticide: Evaluation of some genetically modified acetylcholinesterases from Drosophila melanogaster , 2001 .

[10]  Till T Bachmann,et al.  Design of acetylcholinesterases for biosensor applications. , 2003, Biosensors & bioelectronics.