Structural Characterization of Oligosaccharides Using Maldi-TOF/TOF Tandem Mass Spectrometry

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Abstract

Extensive cross-ring fragmentation ions, which are very informative of the linkages of the monosaccharide residues constituting these molecules, were readily observed in the MALDI-TOF/TOF/MS/MS spectra of oligosaccharides. These ions, in some cases, were more intense than the commonly observed Y and B ions. The A-type ions observed for the simple oligosaccharides allowed the distinction between alpha(1-4)- and alpha(1-6)-linked isobaric structures. The distinction was based not merely on the differences in the type of ions formed, but also on the ion intensities. For example, both alpha(1-4)- and alpha(1-6)-linked isobaric structures produce ions resulting from the loss of approximately 120 m/z units, but with different intensities, as a result of the fact that they correspond to two different ions (i.e., 0,4A- and 2,4A-ions), requiring different energies to be formed. Abundant A- and X-type ions were also observed for high-mannose N-glycans, allowing the determination of linkages. In addition, the high resolution furnished by MALDI-TOF/TOF allowed determination of certain ions that were commonly overlooked by MALDI-TOF or MALDI-magnetic sector instruments as a result of their lower resolution. Moreover, as a result of the fact that MS/MS spectra for parent ions and all fragment ions are acquired under the same experimental conditions, accurate determination of the molar ratios of isomeric glycans in a mixture analyzed simultaneously by MALDI-TOF/TOF tandem MS becomes possible.

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