title: Analysis of glycosaminoglycan-derived, precolumn, 2-aminoacridone-labeled disaccharides with LC-fluorescence and LC-MS detection.
abstract: Glycosaminoglycans (GAGs) possess considerable heterogeneity in average molecular mass, molecular mass range, disaccharide composition and content and position of sulfo groups. Despite recent technological advances in the analysis of GAGs, the determination of GAG disaccharide composition still remains challenging and provides key information required for understanding GAG function. Analysis of GAG-derived disaccharides relies on enzymatic treatment, providing one of the most practical and quantitative approaches for compositional mapping. Tagging the reducing end of disaccharides with an aromatic fluorescent label affords stable derivatives with properties that enable improved detection and resolution. HPLC with on-line electrospray ionization mass spectrometry (ESI-MS) offers a relatively soft ionization method for detection and characterization of sulfated oligosaccharides. GAGs obtained from tissues, biological fluids or cells are treated with various enzymes to obtain disaccharides that are fluorescently labeled with 2-aminoacridone (AMAC) and resolved by different LC systems for high-sensitivity detection by fluorescence, and then they are unambiguously characterized by MS. The preparation and labeling of GAG-derived disaccharides can be performed in ∼1-2 d, and subsequent HPLC separation and on-line fluorescence detection and ESI-MS analysis takes another 1-2 h.
authors: Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
authors: Nicola
authors: N
authors: Volpi
authors: Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
authors: Fabio
authors: F
authors: Galeotti
authors: Departments of Chemistry and Chemical Biology, Chemical and Biological Engineering and Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York, USA.
authors: Bo
authors: B
authors: Yang
authors: Departments of Chemistry and Chemical Biology, Chemical and Biological Engineering and Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York, USA.
authors: Robert J
authors: RJ
authors: Linhardt
mesh_terms: D000609:Aminoacridines / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004187:Disaccharides / Q000032:analysis* / Q000378:metabolism; D005453:Fluorescence; D006025:Glycosaminoglycans / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D013194:Staining and Labeling
chemical_list: D000609:Aminoacridines; D004187:Disaccharides; D006025:Glycosaminoglycans; C072515:2-aminoacridone