Microanalysis of glycosaminoglycan-derived oligosaccharides labeled with afluorophore 2-aminobenzamide by high-performance liquid chromatography: Application to disaccharide composition analysis and exosequencing of oligosaccharides

A series of disaccharides derived from chondroitin sulfate and heparin/hepa ran sulfate were derivatized at their reducing ends with a fluorophore 2-am inobenzamide to develop a sensitive microanalytical method for glycosaminog lycans. The resulting labeled compounds derived from chondroitin sulfate or heparin/heparan sulfate were well-separated and quantified by HPLC equippe d with a fluorescence detector. The detection limit was a low picomole leve l. This method was applied to the analysis of the disaccharide composition of tetra- and hexasaccharides derived from chondroitin sulfate and heparin/ heparan sulfate as well as these glycosaminoglycan polysaccharides. The met hod was also successfully applied to the exosequencing of chondrohexasaccha rides, where the fluorophore-labeled oligosaccharides were degraded exolyti cally from the nonreducing ends using bacterial eliminases. The resultant l abeled fragments were identified by HPLC. (C) 1999 Academic Press.