Fragmentation of negative ions from N-linked carbohydrates. Part 5: Anionic N-linked glycans

The negative ion CID spectra of singly ([M-H](-)) and doubly ([M-2H](2-)) charged sialylated bi-, tri- and tetra-sialylated N-glycans are described with particular reference to ions that define the linkage of the sialic acids. The singly charged ions generally gave prominent deprotonated molecular ions but, because the proton mainly arose from the sialic acid moieties, the fragmentation was not as diagnostic as that of neutral glycans where deprotonation occurs from one of several hydroxyl groups. Nevertheless, ions defining most structural features such as the location of fucose residues and differentiation between triantennary isomers were still present. Ions in the high mass end of the spectra of glycans with alpha 2 -> 6-linked-sialic acids were generally dominated by (0.2)A(7) cleavage products rather than the (2.4)A(7) ions that dominate the spectra of the neutral glycans. The ion at m/z306((0.4)A(2)-CO2) was most useful in defining the alpha 2 -> 6-linkage. The antenna-specific D and [D-18](-) ions were usually present but of low intensity. Glycans with alpha 2 -> 3-linked-sialic acids produced spectra that more closely resembled those of the neutral compounds. Doubly charged ions fragmented mainly to singly charged products following loss of sialic acids. Neutralization by linkage-specific derivatization and consequent formation of [M+anion](-) ions generally restored the abundant antenna-specific fragments. Singly charged sulfated glycans fragmented in a similar manner to the sialylated compounds but the lower end of the spectra were dominated by B-1 (sulfated GaINAc) and B-2 fragments rather than the cross-ring fragments common to the corresponding spectra of the sialylated compounds. (C) 2010 Elsevier B.V. All rights reserved.