Effect of hypochlorite oxidation on cholinesterase-inhibition assay of acetonitrile extracts from fruits and vegetables for monitoring traces of organophosphate pesticides

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  • Kitamura Kentaro
    Faculty of Pharmaceutical Sciences, Tokyo University of Science
  • Maruyama Kaori
    Faculty of Pharmaceutical Sciences, Tokyo University of Science
  • Hamano Sachiko
    Faculty of Pharmaceutical Sciences, Tokyo University of Science
  • Kishi Tomohiro
    Faculty of Pharmaceutical Sciences, Tokyo University of Science
  • Kawakami Tsuyoshi
    Faculty of Pharmaceutical Sciences, Tokyo University of Science Present address: National Institute of Health Sciences
  • Takahashi Yasuo
    Faculty of Pharmaceutical Sciences, Tokyo University of Science
  • Onodera Sukeo
    Faculty of Pharmaceutical Sciences, Tokyo University of Science

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A reproducible method for monitoring traces of cholinesterase (ChE) inhibitors in acetonitrile extracts from fruits and vegetables is described. The method is based on hypochlorite oxidation and ChE inhibition assay. Four common representative samples of produce were selected from a supermarket to investigate the effect of different matrices on pesticides recoveries and assay precision. The samples were extracted with acetonitrile to prepare them for ChE inhibition assays: if necessary, clean-up was performed using dispersive solid-phase extraction for gas chromatography-mass spectrometry (GC/MS) analyses. Chlorine was tested as an oxidising reagent for the conversion of thiophosphorus pesticides (P=S compounds) into their P=O analogues, which have high ChE-inhibiting activity. Chlorine consumption of individual acetonitrile extracts was determined and was strongly dependent on the individual types of fruits and vegetables. After treating the acetonitrile extracts with an excess hypochlorite at 25°C for 15 min, the ChE-inhibiting activities and detection limits for each chlorine-treated pesticide solution were determined. Matrix composition did not interfere significantly with the determination of the pesticides. Enhanced anti-ChE activities leading to low detection limits (ppb levels) were observed for the chlorine-treated extracts that were spiked with chlorpyrifos, diazinon, fenitrothion, and isoxathion. This combination of oxidative derivatisation and ChE inhibition assays was used successfully to monitor and perform semi-quantitative determination of ChE inhibitors in apple, tomato, cucumber, and strawberry samples.

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