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In vitro adduct formation of phosgene with albumin and hemoglobin in human blood

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Author: Noort, D. · Hulst, A.G. · Fidder, A. · Gurp, R.A. van · Jong, L.P.A. de · Benschop, H.P.
Institution: Prins Maurits Laboratorium TNO
Source:Chemical Research in Toxicology, 8, 13, 719-726
Identifier: 235658
doi: doi:10.1021/tx000022z
Keywords: Toxicology · Albumins · Alpha globin · Amino acid sequence · Environmental exposure · Humans · Protein analysis · Protein binding · Protein blood level · Protein determination · Protein synthesis · Radioactivity · Chromatography, High Pressure Liquid · Environmental monitoring · Hemoglobins · Mass Spectrometry · Phosgene · Protein Binding · Hemoglobins · Phosgene, 75-44-5


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 [14C]phosgene (0-750 μM), 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 α-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 μM phosgene.