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Anal Bioanal Chem ; 410(3): 1039-1051, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28971225

RESUMO

Organophosphorus nerve agent (OPNA) adducts formed with human butyrylcholinesterase (HuBuChE) can be used as biomarker of OPNA exposure. Indeed, intoxication by OPNAs can be confirmed by the LC/MS2 analysis of a specific HuBuChE nonapeptide on which OPNAs covalently bind. A fast, selective, and highly sensitive online method was developed to detect sarin and soman adducts in plasma, including immunoextraction by anti-HuBuChE antibodies, pepsin digestion on immobilized enzyme reactors (IMER), and microLC/MS2 analysis of the OPNA adducts. The potential of three different monoclonal antibodies, covalently grafted on sepharose, was compared for the extraction of HuBuChE. The online method developed with the most promising antibodies allowed the extraction of up to 100% of HuBuChE contained in plasma and the digestion of 45% of it in less than 40 min. Moreover, OPNA-HuBuChE adducts, aged OPNA adducts, and unadducted HuBuChE could be detected (with S/N > 2000), even in plasma spiked with a low concentration of OPNA (10 ng mL-1). Finally, the potential of this method was compared to approaches involving other affinity sorbents, already described for HuBuChE extraction. Graphical abstract Online coupling of immunoextraction, digestion, and microliquid chromatography-tandem mass spectrometry for the analysis of organophosphorous nerve agent adducts formed with human butyrylcholinesterase.


Assuntos
Butirilcolinesterase/metabolismo , Substâncias para a Guerra Química/farmacocinética , Inibidores da Colinesterase/sangue , Sarina/sangue , Soman/sangue , Butirilcolinesterase/sangue , Substâncias para a Guerra Química/metabolismo , Inibidores da Colinesterase/metabolismo , Cromatografia de Afinidade/métodos , Cromatografia Líquida/métodos , Humanos , Sarina/análogos & derivados , Sarina/metabolismo , Soman/análogos & derivados , Soman/metabolismo , Espectrometria de Massas em Tandem/métodos
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