Title
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
Author
Noort, D.
Fidder, A.
Hulst, A.G.
de Jong, L.P.A.
Benschop, H.P.
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
Publication year
2000
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.
Subject
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
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TNO identifier
346139
ISSN
0260-437X
Source
Journal of Applied Toxicology, 20 (Suppl. 1), S187-S192
Document type
article