title: In vitro adduct formation of phosgene with albumin and hemoglobin in human blood.
abstract: The development of procedures for retrospective detection and quantitation of exposure to phosgene, based on adducts to hemoglobin and albumin, is described. Upon incubation of human blood with [(14)C]phosgene (0-750 microM), a significant part of radioactivity (0-13%) became associated with globin and albumin. Upon Pronase digestion of globin, one of the adducts was identified as the pentapeptide O=C-(V-L)-S-P-A, representing amino acid residues 1-5 of alpha-globin, with a hydantoin function between N-terminal valine and leucine. Micro-LC/tandem MS analyses of tryptic as well as V8 protease digests identified one of the adducts to albumin as a urea resulting from intramolecular bridging of lysine residues 195 and 199. The adducted tryptic fragment could be sensitively analyzed by means of micro-LC/tandem MS with multiple-reaction monitoring (MRM), enabling the detection in human blood of an in vitro exposure level of >/=1 microM phosgene.
authors.affiliation: Department of Chemical Toxicology and Department of Analysis of Toxic and Explosive Substances, TNO Prins Maurits Laboratory, P.O. Box 45, 2280 AA Rijswijk, The Netherlands. noortd@pml.tno.nl
authors.forename: D
authors.initials: D
authors.lastname: Noort
authors.forename: A G
authors.initials: AG
authors.lastname: Hulst
authors.forename: A
authors.initials: A
authors.lastname: Fidder
authors.forename: R A
authors.initials: RA
authors.lastname: van Gurp
authors.forename: L P
authors.initials: LP
authors.lastname: de Jong
authors.forename: H P
authors.initials: HP
authors.lastname: Benschop
mesh_terms: D000418:Albumins / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004781:Environmental Exposure; D004784:Environmental Monitoring / Q000379:methods; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D010705:Phosgene / Q000378:metabolism* / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects
chemical_list: D000418:Albumins; D006454:Hemoglobins; D010705:Phosgene