Sensitive detection of organophosphorus pesticides using a needle type amperometric acetylcholinesterase-based bioelectrode. Thiocholine electrochemistry and immobilised enzyme inhibition

J Enzyme Inhib Med Chem. 2002 Apr;17(2):107-15. doi: 10.1080/14756360290026469.

Abstract

An acetylcholinesterase (AChE) based amperometric bioelectrode for a selective detection of low concentrations of organophosphorus pesticides has been developed. The amperometric needle type bioelectrode consists of a bare cavity in a PTFE isolated Pt-Ir wire, where the AChE was entrapped into a photopolymerised polymer of polyvinyl alcohol bearing styrylpyridinium groups (PVA-SbQ). Cyclic voltammetry, performed at Pt and AChE/Pt disk electrodes, confirmed the irreversible, monoelectronic thiocholine oxidation process and showed that a working potential of +0.410 V vs. Ag/AgCl, KCl(sat) was suitable for a selective and sensitive amperometric detection of thiocholine. The acetylthiocholine detection under enzyme kinetic control was found in the range of 0.01-0.3 U cm(-2) of immobilised AChE. The detection limit, calculated for an inhibition ratio of 10%, was found to reach 5 microM for dipterex and 0.4 microM for paraoxon. A kinetic analysis of the AChE-pesticide interaction process using Hanes-Woolf or Lineweaver-Burk linearisations and secondary plots allowed identification of the immobilised enzyme inhibition process as a mixed one (non/uncompetitive) for both dipterex and paraoxon. The deviation from classical Michaelis Menten kinetics induced from the studied pesticides was evaluated using Hill plots.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcholinesterase / metabolism*
  • Animals
  • Cholinesterase Inhibitors / chemistry*
  • Electrochemistry / methods
  • Electrodes
  • Electrophorus
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / pharmacology*
  • Enzymes, Immobilized / antagonists & inhibitors*
  • Insecticides / chemistry
  • Insecticides / pharmacology*
  • Kinetics
  • Paraoxon / analysis
  • Paraoxon / pharmacology
  • Sensitivity and Specificity
  • Thiocholine / chemistry*
  • Trichlorfon / analysis
  • Trichlorfon / pharmacology

Substances

  • Cholinesterase Inhibitors
  • Enzyme Inhibitors
  • Enzymes, Immobilized
  • Insecticides
  • Thiocholine
  • Trichlorfon
  • Acetylcholinesterase
  • Paraoxon