Chitooligosaccharides with degrees of polymerization (DPs) 2-12 were successfully separated and characterized using LC-MS/MS analysis with an amine column and a quadrupole ion trap analyzer. Major MS peaks for each chromatographic peak originated from deacetylated chitooligosaccharides while N-acetylated chitooligosaccharides were observed as minor peaks. The ratio of acetylation per glucosamine unit for each chitooligomer was higher for oligomers with higher DPs. The MS/MS spectra of chitooligosaccharides were used to characterize the chitooligosaccharide fragmentation patterns. To the best of our knowledge, this is the first report of on-line LC-MS/MS analysis of chitooligosaccharides with DP higher than 6.
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