Accession Number : ADP008768


Title :   Mass Spectrometric Detection of Sulfur Mustard Adducts to Proteins and DNA: Dosimetry of Exposure to Sulfur Mustard


Corporate Author : PRINS MAURITS LABORATORIUM TNO RIJSWIJK(NETHERLANDS)


Personal Author(s) : Fidder, A. ; Noort, D. ; Hulst, A. G. ; de Jong, L. P. ; Benschop, H. P.


Full Text : http://www.dtic.mil/dtic/tr/fulltext/u2/p008768.pdf


Report Date : 13 MAY 1993


Pagination or Media Count : 9


Abstract : Immunochemical detection methods have been developed for analysis of protein and DNA adducts of sulfur mustard within our studies performed in collaboration with TNO-Medical Biological Laboratory. In order to validate immunochemical determinations, mass spectrometric detection methods are being developed as an independent, highly sensitive technique having an almost absolute specificity. A method has been developed for specific cleavage of the alkylated N-terminal valine in the alpha-chain of human hemoglobin exposed to sulfur mustard, by using the modified Edman reagent pentafluorophenyl isothiocyanate. After subsequent reaction with heptafluorobutyric anhydride, the derivatized alkylated valine is determined by gas chromatography/negative chemical ionization-mass spectrometry (GC/NCI-MS). In a second approach, a procedure for analysis of the major amino acid adduct formed upon alkylation of hemoglobin with sulfur mustard will be developed. To this end, the protein is alkylated with 35S-sulfur mustard and is subsequently digested by a proteolytic enzyme. After appropriate derivatization of the (alkylated) amino acids in the digest, HPLC analyses with radiometric detection and spiking with synthesized reference adducts and GC/MS analyses are performed in order to show which alkylated amino acids have been formed, and survive digestion. Furthermore, adducted peptides were identified by LC-MS Analysis in a trypsin digest of the alkylated globin.


Descriptors :   *PROTEINS , *DEOXYRIBONUCLEIC ACIDS , *MUSTARD AGENTS , *EXPOSURE(PHYSIOLOGY) , *IMMUNOCHEMISTRY , *DOSIMETRY , DETECTION , HUMANS , CHEMICALS , PEPTIDES , ENZYMES , ANHYDRIDES , SPECTROMETRY , CHAINS , IONIZATION , ALKYLATION , SULFUR , HEMOGLOBIN , DETERMINATION , GAS CHROMATOGRAPHY , TRYPSIN , AMINO ACIDS , CHROMATOGRAPHY , CLEAVAGE , PEPTIDE HYDROLASES , MASS SPECTROMETRY , HYDROLASES , TERMINALS , APPROACH , BIOLOGICAL LABORATORIES , PROTEINS(SIMPLE).


Subject Categories : BIOCHEMISRTY
      TOXICOLOGY
      CHEMICAL, BIOLOGICAL AND RADIOLOGICAL WARFARE


Distribution Statement : APPROVED FOR PUBLIC RELEASE