The use of capillary gas chromatography-mass spectrometry for identification of radiation-induced DNA base damage and DNA base-amino acid cross-links.
Application of capillary gas chromatography-mass spectrometry (GC-MS) to isolation and identification of radiation-induced DNA base damage including DNA base-amino acid crosslinks is reported. gamma-Irradiated samples of thymine (thy), thymidine (dT), thymidine-5'-monophosphate (pdT), cytosine(cyt),2'-deoxycytidine (dC), 2'-deoxycytidine-5'-monophosphate (pdC), and mixtures of thy, dT and pdT with tyrosine were used as model systems. Trimethylsilylation was used as the derivatization method. Samples containing nucleosides and nucleotides were first subjected to hydrolysis with formic acid or hydrochloric acid to remove sugar or sugar-phosphate moieties, then trimethylsilylated and analyzed by GC-MS. Trimethylsilyl derivatives of radiation-induced monomeric and dimeric products of the model systems mentioned above were shown to have excellent GC properties and easily interpretable mass spectra. The presence of the molecular ion (M+.) and a characteristic (M-CH3)+ ion in the mass spectra facilitated structural elucidation. The complementary use of tert.-butyldimethylsilyl derivatives was also demonstrated. These derivatives provided an intense characteristic (M-57)+ ion in their mass spectra, which may be used to corroborate the molecular weight and to monitor the corresponding compounds in a complex mixture by means of selected-ion monitoring. All gas chromatograms and mass spectra obtained are discussed in detail.
D000596:Amino Acids / Q000528:radiation effects*; D001482:Base Composition / Q000528:radiation effects; D055598:Chemical Phenomena; D002621:Chemistry; D002849:Chromatography, Gas / Q000379:methods; D003596:Cytosine / Q000528:radiation effects; D004247:DNA / Q000032:analysis / Q000528:radiation effects*; D003841:Deoxycytidine / Q000528:radiation effects; D005561:Formates / Q000528:radiation effects; D005720:Gamma Rays; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006851:Hydrochloric Acid / Q000528:radiation effects; D006868:Hydrolysis; D013936:Thymidine / Q000528:radiation effects; D013938:Thymidine Monophosphate / Q000528:radiation effects; D013941:Thymine / Q000528:radiation effects; D014443:Tyrosine / Q000528:radiation effects
D000596:Amino Acids; D005561:Formates; D003841:Deoxycytidine; C030544:formic acid; D013938:Thymidine Monophosphate; D014443:Tyrosine; D003596:Cytosine; D004247:DNA; D013941:Thymine; D006851:Hydrochloric Acid; D013936:Thymidine