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We propose a novel, bi-dimensional analysis of partially  N -acetylated chitosan oligosaccharides based on gas phase Infra-Red spectroscopy integrated to mass spectrometry (MS). By providing simultaneously MS and IR fingerprints, this approach combines the advantages of MS with the refined structural detail offered by gas phase spectroscopy and provides robust signatures for the rapid discrimination of the patterns of  N -acetylation. Four mono- N -deacetylated and two doubly- N -deacetylated chitosan tetramer standards with well-defined patterns of acetylation were produced and analyzed by IR integrated to MS. We show that each sequence displays a unique combination of MS and IR fingerprints, thus offering a rapid diagnostic for the pattern of acetylation without the need for reducing end labeling.

Discrimination of patterns of N -acetylation in chitooligosaccharides by gas phase IR spectroscopy integrated to mass spectrometry