Characterization of cytosine nucleosides by mass spectrometry

The mass spectra of most nucleosides of cytosine exhibit molecular ions of low abundance, rearranged ions base + 2H in greater abundance than the companion species base + H, and a characteristic and abundant ion which corresponds to base + C2HO from the sugar. Deuterium, oxygen-18 and substituent labeling showed the presence of C-1′, 2′, H-2′ and O-4′ in the latter ion, which led to a proposed mechanism of formation involving opening of the sugar ring and recyclization between O2 and C-2′ with expulsion of water. Primary driving force for the reaction is due to resonance stabilization promoted by unshared electrons from N4, which results in diminished abundance in the case of imino isomers and derivatives which contain electron-withdrawing groups at N4, and enhanced abundance resulting from methylation at N4. Data for 24 nucleosides are presented and discussed in terms of the determination of nucleoside structure.