Separation and Identification of Glycan Anomers Using Ultrahigh-Resolution Ion Mobility Spectrometry and Cryogenic Ion Spectroscopy

The analysis of carbohydrates, or glycans, is challenging for established structure-sensitive gas-phase methods. The multitude of possible stereo-, regio- and structural isomers make them substantially more complex to ana-lyze than DNA or proteins, and no one method is currently able to fully resolve them. While the combination of tandem mass spectrometry (MS) and ion mobility spectrometry (IMS) have made important inroads in gly-can analysis, in many cases this approach is still not able to identify the precise isomeric form. To advance the techniques available for glycan analysis we employ two important innovations. First, we perform ultrahigh-resolution mobility separation using structures for lossless ion manipulations (SLIM) for isomer separation and pre-selection. We then complement this IMS-MS stage with a cryogenic IR spectroscopic dimension, since a glycan's vibrational spectrum provides a fingerprint that is extremely sensitive to the precise isomeric form. Using this unique approach in conjunction with oxygen-18 isotopic labelling, we show on a range of disaccha-rides how the two  and  anomers that every reducing glycan adopts in solution can be readily separated by mobility and identified based on their IR spectra. In addition to highlighting the power of our technique to detect minute differences in the structure of isomeric carbohydrates, these results provide the means to determine if and when anomericity is retained during collision-induced dissociation (CID) of larger glycans.


Published in:
Journal of the American Society for Mass Spectrometry
Year:
2019
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 Record created 2019-09-05, last modified 2019-12-05

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