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They are ubiquitous molecules, which are involved in a wide range of biological processes. Despite being structurally simple at first glance, with a repeating backbone of alternating hexuronic acid and hexosamine dimers, GAGs display a highly complex structure, which predominantly results from their heterogeneous sulfation patterns. The commonly applied method for compositional analysis of all GAGs is \u201cdisaccharide analysis.\u201d In this process, GAGs are enzymatically depolymerized into disaccharides, derivatized with a fluorescent label, and then analysed through liquid chromatography. The limiting factor in the high throughput analysis of GAG disaccharides is the time\u2010consuming liquid chromatography. To address this limitation, we here utilized trapped ion mobility\u2010mass spectrometry (TIM\u2010MS) for the separation of isomeric GAG disaccharides, which reduces the measurement time from hours to a few minutes. A full set of disaccharides comprises twelve structures, with eight possessing isomers. Most disaccharides cannot be differentiated by TIM\u2010MS in underivatized form. Therefore, we developed chemical modifications to reduce sample complexity and enhance differentiability. Quantification is performed using stable isotope labelled standards, which are easily available due to the nature of the performed modifications.<\/jats:p>","DOI":"10.1002\/chem.202400783","type":"journal-article","created":{"date-parts":[[2024,4,17]],"date-time":"2024-04-17T05:48:37Z","timestamp":1713332917000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Ion Mobility Mass Spectrometry\u2010Based Disaccharide Analysis of Glycosaminoglycans"],"prefix":"10.1002","volume":"30","author":[{"given":"Lukasz","family":"Polewski","sequence":"first","affiliation":[{"name":"Institute of Chemistry and Biochemistry Freie Universit\u00e4t Berlin  14195 Berlin Altensteinstra\u00dfe 23a Germany"},{"name":"Department of Molecular Physics Fritz-Haber-Institut der Max-Planck-Gesellschaft  14195 Berlin Faradayweg 4\u20136 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