Synthesis and mass spectrometric identification of the major amino acid adducts formed between sulphur mustard and haemoglobin in human blood.

Autor: Noort, Daan, Hulst, Albert G., Trap, H. C., de Jong, Leo P. A., Benschop, Hendrik P.
Zdroj: Archives of Toxicology; Jan1997, Vol. 71 Issue 3, p171-178, 8p
Abstrakt: As part of a program to develop methods for the verification of alleged exposure to sulphur mustard, we synthesized and characterized three amino acid adducts presumably formed by alkylation of haemoglobin: 4-(2-hydroxyethylthioethyl)- l-aspartate, 5-(2-hydroxyethylthioethyl)- l-glutamate and N1- and N3-(2-hydroxyethylthioethyl)- l-histidine. Suitable derivatization methods for GC/MS analysis were developed for these adducts as well as for the cysteine and the N-terminal valine adduct. Incubation of human blood with [35S]sulphur mustard in vitro followed by acidic hydrolysis of isolated globin and derivatization with Fmoc-Cl afforded three major radioactive peaks upon HPLC analysis, one of which coeluted with the synthetic Fmoc derivative of N1/N3-(2-hydroxyethylthioethyl)- l-histidine. After pronase digestion of globin the adducts of histidine, glutamic acid, aspartic acid, cysteine and N-terminal valine could be tentatively identified and quantitated. Final identification was obtained from GC/MS analysis. The most abundant adduct, N1/N3-(2-hydroxyethylthioethyl)- l-histidine, could not be sensitively analysed by GC/MS. A convenient LC-tandem MS procedure was developed for this compound, enabling the detection of exposure of human blood to 10␣μM sulphur mustard in vitro. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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