In vitro adduct formation of phosgene with albumin and hemoglobin in human blood.
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.
Noort
D
D
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
Hulst
A G
AG
Fidder
A
A
van Gurp
R A
RA
de Jong
L P
LP
Benschop
H P
HP
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
D000418:Albumins; D006454:Hemoglobins; D010705:Phosgene