Evidence of exposure to organophosphorus toxicants by detection of the propionylated butyrylcholinesterase-derived nonapeptide-adduct as a novel biomarker. (June 2021)

• Record Type: Journal Article
• Authors: John, Harald; Richter, Annika; Siegert, Markus; Eyer, Florian; Thiermann, Horst
• Is Part Of: Forensic science international. Volume 323(2021)
• Publication Date: 2021-06
• Subjects: BChE-adducts -- Derivatization -- LC MS/MS -- Nerve agents -- Pesticides -- Propionic anhydride -- Verification; ACN acetonitrile -- BChE butyrylcholinesterase -- BioPT bioproficiency test of the OPCW -- CID collision-induced dissociation -- CVX Chinese VX -- d3-Atr triple deuterated atropine -- DEP diethyl phosphate -- DMP dimethyl phosphate -- ESI electrospray ionization -- FA formic acid -- GA tabun -- GB sarin -- GD soman -- GF cyclosarin -- HR high-resolution -- HSA human serum albumin -- ICU intensive care unit -- IMS immunomagnetic separation -- IMPA isopropyl methyl phosphonic acid -- µLC micro liquid chromatography -- LOD lower limit of detection -- LOI lower limit of identification -- MS mass spectrometry -- MS/MS tandem-mass spectrometry -- NA nerve agent -- NP nonapeptide -- OP organophosphorus -- OPCW Organisation for the Prohibition of Chemical Weapons -- PA-NP propionylated nonapeptide -- Q1 – Q4 qualifying product ions 1–4 -- rf molecular response factor -- RT room temperature -- RVX Russian VX -- SRM selected reaction monitoring -- UF ultrafiltration; Medical jurisprudence -- Periodicals; Chemistry, Forensic -- Periodicals; Forensic Medicine -- Periodicals; Médecine légale -- Périodiques; Chimie légale -- Périodiques; Gerechtelijke geneeskunde; Gerechtelijke chemie; Gerechtelijke psychiatrie; Chemistry, Forensic; Medical jurisprudence; Electronic journals; Periodicals; Electronic journals; 614.1
• DOI: 10.1016/j.forsciint.2021.110818
• Languages: English
• ISSNs: 0379-0738

Abstract

Abstract: Organophosphorus (OP) nerve agents represent a class of highly toxic chemical warfare agents banned by the Chemical Weapons Convention. Nevertheless, in the past few years they have been used repeatedly for warfare, assassination and attempted murder. In addition, the chemically related OP pesticides were frequently used for suicide and may be deployed for terroristic attacks. Therefore, sensitive and selective bioanalytical methods are indispensable to investigate biological specimens as pieces of evidence to prove poisoning. OP agents form long-lived covalent reaction products (adducts) with endogenous proteins like human serum albumin (HSA) and butyrylcholinesterase (BChE). The adducted nonapeptide (NP) obtained by proteolysis of the BChE-adduct is one of the most sensitive and important biomarkers. We herein present a novel class of NP-adducts propionylated at its N-terminal phenylalanine residue (F 195 ). The biomarker derivative is produced by addition of propionic anhydride to the NP-adduct inducing its quantitative conversion in aqueous buffer within 5 min at room temperature. Afterwards the mixture is directly analyzed by micro-liquid chromatography-electrospray ionization tandem-mass spectrometry (µLC-ESI MS/MS). The sensitivity of the method is comparable to that of the non-derivatized NP-adduct. These characteristics make the method highly beneficial for forensic analysis especially in cases in which the OP agent does not form adducts with HSA that are typically targeted as a second biomarker of exposure. This novel procedure was successfully applied to nerve agent-spiked samples sent by the Organisation for the Prohibition of Chemical Weapons (OPCW) as well as to plasma samples of real cases of pesticide poisoning.