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.
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.
Noort
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D
Department of Chemical Toxicology, Rijswijk, The Netherlands.
Fidder
A
A
Hulst
A G
AG
de Jong
L P
LP
Benschop
H P
HP
D000418:Albumins / Q000032:analysis* / Q000737:chemistry; D002250:Carbon Radioisotopes; D018929:Cell Culture Techniques; D002853:Chromatography, Liquid; D003937:Diagnosis, Differential; D006454:Hemoglobins / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D007633:Keratins / Q000032:analysis* / Q000737:chemistry; D013058:Mass Spectrometry; D009151:Mustard Gas / Q000506:poisoning*; D011041:Poisoning / Q000175:diagnosis; D012016:Reference Values; D012867:Skin / Q000187:drug effects / Q000473:pathology
D000418:Albumins; D002250:Carbon Radioisotopes; D006454:Hemoglobins; D007633:Keratins; D009151:Mustard Gas