title: GC-MS and LC-MS analysis of nerve agents in body fluids: intra-laboratory verification test using spiked plasma and urine samples.
abstract: The purpose of this study was to check the applicability of different analytical methods for the identification of unknown nerve agents in human body fluids. Plasma and urine samples were spiked with nerve agents (plasma) or with their metabolites (urine) or were left blank. Seven random samples (35% of all samples) were selected for the verification test. Plasma was worked up for unchanged nerve agents and for regenerated nerve agents after fluoride-induced reactivation of nerve agent-inhibited butyrylcholinesterase. Both extracts were analysed by GC-MS. Metabolites were extracted from plasma and urine, respectively, and were analysed by LC-MS. The urinary metabolites and two blank samples could be identified without further measurements, plasma metabolites and blanks were identified in six of seven samples. The analysis of unchanged nerve agent provided five agents/blanks and the sixth agent after further investigation. The determination of the regenerated agents also provided only five clear findings during the first screening because of a rather noisy baseline. Therefore, the sample preparation was extended by a size exclusion step performed before addition of fluoride which visibly reduced baseline noise and thus improved identification of the two missing agents. The test clearly showed that verification should be performed by analysing more than one biomarker to ensure identification of the agent(s).
authors.affiliation: Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, D-80937 München, Germany. mariannekoller@bundeswehr.org
authors.forename: Marianne
authors.initials: M
authors.lastname: Koller
authors.forename: Christian
authors.initials: C
authors.lastname: Becker
authors.forename: Horst
authors.initials: H
authors.lastname: Thiermann
authors.forename: Franz
authors.initials: F
authors.lastname: Worek
mesh_terms: D002091:Butyrylcholinesterase; D002619:Chemical Warfare Agents / Q000032:analysis* / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods*; D005459:Fluorides; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D009943:Organophosphorus Compounds / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000652:urine; D015203:Reproducibility of Results; D052616:Solid Phase Extraction
chemical_list: D002619:Chemical Warfare Agents; D009943:Organophosphorus Compounds; D002091:Butyrylcholinesterase; D005459:Fluorides