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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

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Author: Noort, D. · Fidder, A. · Hulst, A.G. · Jong, L.P.A. de · Benschop, H.P.
Type:article
Date:2000
Institution: Department of Chemical Toxicology, TNO Prins Maurits Laboratory, PO Box 45, 2280 AA Rijswijk, Netherlands Department of Analysis of Toxic and Explosive Substances, TNO Prins Maurits Laboratory, PO Box 45, 2280 AA Rijswijk, Netherlands
Source:Journal of Applied Toxicology, Suppl. 1, 20, S187-S192
Identifier: 346139
Keywords: Adducts · Albumin · Dosimetry · Haemoglobin · Keratin · Mass spectrometry · Sulfur mustard · Albuminoid · Carbon · Diagnostic agent · Hemoglobin · Keratin · Mustard gas · Article · Chemistry · Culture technique · Differential diagnosis · Drug effect · Human · Intoxication · Liquid chromatography · Mass spectrometry · Pathology · Reference value · Skin · Albumins · Carbon Radioisotopes · Cell Culture Techniques · Chromatography, Liquid · Diagnosis, Differential · Hemoglobins · Humans · Mass Spectrometry · Poisoning · Reference Values · Skin · Carbon Radioisotopes · Keratins, 68238-35-7 · Mustard Gas, 505-60-2

Abstract

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°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: ≥15 pg absolute and 1 μM 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. Copyright © 2000 John Wiley & Sons, Ltd.