TY - JOUR
T1 - Disaccharide analysis of glycosaminoglycan mixtures by ultra-high-performance liquid chromatography-mass spectrometry
AU - Yang, Bo
AU - Chang, Yuqing
AU - Weyers, Amanda M.
AU - Sterner, Eric
AU - Linhardt, Robert J.
N1 - Funding Information:
The authors are grateful for funding from the National Institutes of Health in the forms of grants GM38060 , GM090127 , HL096972 , HL101721 and the NY State Stem Cell Foundation N08G-264 .
PY - 2012/2/17
Y1 - 2012/2/17
N2 - Glycosaminoglycans are a family of polysaccharides widely distributed in all eukaryotic cells. These polyanionic, linear chain polysaccharides are composed of repeating disaccharide units that are often differentially substituted with sulfo groups. The diversity of glycosaminoglycan structures in cells, tissues and among different organisms reflect their functional an evolutionary importance. Glycosaminoglycan composition and structure also changes in development, aging and in disease progression, making their accurate and reliable analysis a critical, albeit, challenging endeavor. Quantitative disaccharide compositional analysis is one of the primary ways to characterize glycosaminoglycan composition and structure and has a direct relationship with glycosaminoglycan biological functions. In this study, glycosaminoglycan disaccharides, prepared from heparan sulfate/heparin, chondroitin sulfate/dermatan sulfate and neutral hyaluronic acid using multiple polysaccharide lyases, were fluorescently labeled with 2-aminoacridone, fractionated into 17 well-resolved components by reverse-phase ultra-performance liquid chromatography, and analyzed by electrospray ionization mass spectrometry. This analysis was successfully applied to cell, tissue, and biological fluid samples for the picomole level detection of glycosaminoglycan composition and structure.
AB - Glycosaminoglycans are a family of polysaccharides widely distributed in all eukaryotic cells. These polyanionic, linear chain polysaccharides are composed of repeating disaccharide units that are often differentially substituted with sulfo groups. The diversity of glycosaminoglycan structures in cells, tissues and among different organisms reflect their functional an evolutionary importance. Glycosaminoglycan composition and structure also changes in development, aging and in disease progression, making their accurate and reliable analysis a critical, albeit, challenging endeavor. Quantitative disaccharide compositional analysis is one of the primary ways to characterize glycosaminoglycan composition and structure and has a direct relationship with glycosaminoglycan biological functions. In this study, glycosaminoglycan disaccharides, prepared from heparan sulfate/heparin, chondroitin sulfate/dermatan sulfate and neutral hyaluronic acid using multiple polysaccharide lyases, were fluorescently labeled with 2-aminoacridone, fractionated into 17 well-resolved components by reverse-phase ultra-performance liquid chromatography, and analyzed by electrospray ionization mass spectrometry. This analysis was successfully applied to cell, tissue, and biological fluid samples for the picomole level detection of glycosaminoglycan composition and structure.
KW - Chondroitin sulfate
KW - Disaccharide analysis
KW - Fluorescently labeling
KW - Heparin
KW - Hyaluronan
KW - LC-MS
UR - http://www.scopus.com/inward/record.url?scp=84856229594&partnerID=8YFLogxK
U2 - 10.1016/j.chroma.2011.12.063
DO - 10.1016/j.chroma.2011.12.063
M3 - Article
C2 - 22236563
AN - SCOPUS:84856229594
SN - 0021-9673
VL - 1225
SP - 91
EP - 98
JO - Journal of Chromatography A
JF - Journal of Chromatography A
ER -