Abstract
Recent events have shown that organophosphorus nerve agents (OPNAs) are a serious threat. Cholinesterase inhibition by OPNAs results in acetylcholine accumulation, a cholinergic crisis leading to death if untreated. Efficacy assessment of new medical countermeasures against OPNAs relies on translational animal models. We developed a swine model of percutaneous VX intoxication and a simple plate reader-based enzymatic method to quantify plasmatic VX over time. Juvenile pigs anesthetized with sevoflurane were poisoned with a single supralethal (n = 5; 1200 μg/kg) or sublethal (n = 6; 320 μg/kg) percutaneous dose of VX. These intoxicated animals were compared to 7 control animals. Repeated blood sampling was performed up to 6 h post-intoxication. Blood cholinesterase activities were measured using the Ellman assay. Nanomolar plasma concentrations of VX were measured by exogenous butyrylcholinesterase added to an aliquot of plasma. As expected, we observed a steady increase in plasma concentration of VX over time concomitant to a decrease in blood cholinesterase activities for all intoxicated pigs. Despite the simplicity of the enzymatic method, the results obtained are in good agreement with those of the liquid chromatography–mass spectrometry method. This method is also applicable to other OPNAs such as novichoks with minor adaptations.
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Abbreviations
- AChE:
-
Acetylcholinesterase
- ATC:
-
Acetylthiocholine
- BChE:
-
Butyrylcholinesterase
- BTC:
-
Butyrylthiocholine
- ChE:
-
Total cholinesterase
- DTNB:
-
5,5′-Dithiobis-2-nitrobenzoic acid
- E:
-
Enzyme
- EOPNA:
-
Inhibited enzyme
- hBChE:
-
Human butyrylcholinesterase
- IV:
-
Intravenous
- ki:
-
Bimolecular rate constant of inhibition
- LC–MS/MS:
-
Liquid chromatography–mass spectrometry
- OPNA:
-
Organophosphorus-nerve agents
- PC:
-
Percutaneous
- VX:
-
O-Ethyl S-diisopropylaminomethyl methylphosphonothiolate
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Acknowledgements
We are thankful to the French Ministry of Armed Forces for sustained funding of biomedical research under grants by the Direction Générale de l’Armement and Service de Santé des Armées (Grant PDH-2-NBC-4-C-4209 and PDH-2-NBC-4-C-4212).
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Fémy, F., Meesemaecker, G., Belverge, N. et al. Toxicokinetics of plasmatic VX in a swine model: comparison of a simple enzymatic titration method with a mass spectrometry method. Arch Toxicol 97, 421–428 (2023). https://doi.org/10.1007/s00204-022-03408-w
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DOI: https://doi.org/10.1007/s00204-022-03408-w