Diagnosis and dosimetry of exposure to sulfur mustard: development of a standard operating procedure for mass spectrometric analysis of haemoglobin adducts: exploratory research on albumin and keratin adducts.

Autor: Noort D; Department of Chemical Toxicology, Rijswijk, The Netherlands., Fidder A, Hulst AG, de Jong LP, Benschop HP
Jazyk: angličtina
Zdroj: Journal of applied toxicology : JAT [J Appl Toxicol] 2000 Dec; Vol. 20 Suppl 1, pp. S187-92.
DOI: 10.1002/1099-1263(200012)20:1+<::aid-jat676>3.0.co;2-f
Abstrakt: Experiments were carried out to develop a standard operating procedure for analysis of sulfur mustard adducts to the N-terminal valine in haemoglobin and to explore adduct formation with albumin and keratin. In the first approach, gas chromatography-negative chemical ionization/mass spectrometry (GC-NCI/MS) of the thiohydantoin sample subsequent to the modified Edman degradation was performed using a thermodesorption/cold trap (TCT) injection technique (detection limit for in vitro exposure of human blood to sulfur mustard: 30 nM). In the second approach, the crude thiohydantoin sample was purified by solid-phase extraction procedures. In the third approach, the procedure was shortened significantly by performing the Edman degradation for 2 h at 60 degrees C. Upon exposure of human blood to various concentrations of [14C]sulfur mustard, ca. 20% was covalently bound to albumin. One of the tryptic fragments (T5 containing an alkylated cysteine (HETE-(A-L-V-L-I-A-F-A-Q-Y-L-Q-Q-C-P-F-E-D-H-V-K); MW 2536 Da) could be detected sensitively with liquid chromatography/tandem mass spectrometry analysis (detection limits: > or =15 pg absolute and 1 microM for in vitro exposure of human blood). Upon exposure of human callus (suspensions in 0.9% NaCl; 500 mg ml(-1)) to various concentrations of [14C]sulfur mustard we found 15-20% of the added radioactivity covalently bound to keratin. Upon incubation with base, 80% of the bound radioactivity was split off as [14C]thiodiglycol. This result opens the way for sensitive mass spectrometric detection of sulfur mustard exposure of skin by gas chromatography/mass spectrometry of (derivatized) thiodiglycol.
Databáze: MEDLINE
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