HPLC-MS Characterization of Tissue-Derived Heparan Sulfate and Chondroitin Sulfate

Glycosaminoglycans (GAGs) are built up of repeating disaccharide units resulting in long, linear polysaccharide chains. In most classes of GAGs, sulfation and epimerization complicate the structure of the chain and influence biochemical functions. The most widespread way of their investigation by instrumental analytical techniques is to degrade them into the constituent disaccharide building blocks, followed by capillary electrophoresis or high-performance liquid chromatography (HPLC) separation. The analysis of GAG disaccharides with varying sulfation degrees poses a real challenge both from chromatographic and mass spectrometric (MS) points of view. This necessitates the constant improvement of their analytical methodology. In this chapter, an optimized workflow will be discussed for the sample preparation and subsequent HPLC-MS characterization of tissue-derived chondroitin sulfate and heparan sulfate.