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Advances in Derivatization Techniques Enabled by DABCO for Novichok Agent Analysis in Biofluids Using LC-MS

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ANALYTICAL CHEMISTRY
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AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.3c02775

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A novel method for analyzing NV hydrolysates using LC-MS/MS enabled by PFB derivatization followed by reaction with DABCO was developed, resulting in improved limit of detection compared to previous methods. Simple pretreatment using DABCO and filtration allowed for LC-MS/MS analysis after reaction with PFBBr. The use of PAM significantly enhanced the detection of NV hydrolysates, although PAM is not a proven antidote for NV.
The characterization of Novichoks (NVs), a new group of nerve agents that have been implicated in two recent poisonings, has not been extensively conducted. Here, we present a novel method for analyzing NV hydrolysates using liquid chromatography-tandem mass spectrometry (LC-MS/MS) enabled by pentafluorobenzyl (PFB) derivatization followed by reaction with 1,4-diazabicyclo[2.2.2]octane (DABCO). This approach enabled efficient, simultaneous screening of six NV hydrolysates, with 1-2 orders improvement in the limit of detection in relation to that achieved through previous methods. A straightforward pretreatment using DABCO and filtration was employed for biological samples, mitigating instrument damage and allowing LC-MS/MS after a reaction with highly hydrophobic PFB bromide (PFBBr). In addition, the use of pralidoxime (PAM) significantly enhanced the detection of NV hydrolysates from NV-surrogate-spiked serum. While PAM is not a proven NV antidote, its effectiveness as an analytical reagent to aid in the detection of NV hydrolysates was demonstrated for the first time. Understanding the proposed mechanism of DABCO-mediated derivatization reagent removal in this research could broaden the range of compounds amenable to derivatization LC, thereby enhancing the capabilities of conventional derivatization techniques.

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