Characterisation of inhibitors of acetylcholinesterase by an automated amperometric flow-injection system

Abstract Acetylcholinesterase was immobilised on magnetic particles and integrated in a flow-injection system via a magnetic reactor. Enzyme activity was determined amperometrically using acetylthiocholine chloride as enzyme substrate. This system was applied to enzyme inhibition tests. Inhibition constants and lower detection limits were determined for carbofuran, paraoxon, malaoxon and paraoxon-methyl. The resulting data were compared to those obtained with a photometric test, i.e. the determination of thiocholine via its reaction with the Ellman's reagent 5,5'-dithio-bis-2-nitrobenzoic acid using the same flow system. As they correlated well to those reported for the native enzyme the flow-injection analysis device can be applied to automated determination and characterisation of enzyme inhibitors.