Comparison of separation modes of high-performance liquid chromatography for the analysis of glycoprotein- and proteoglycan-derived oligosaccharides.
Porous graphitised carbon (PGC) has been explored for high-performance liquid chromatography (HPLC) of mono- and di-saccharides released from proteoglycans and of fluorescently labelled oligosaccharide derivatives for high-sensitivity detection. Sulphated oligosaccharides show good retention and separation behaviour on PGC-HPLC, and compared to anion-exchange or reversed-phase ion-pair chromatography the chromatography is carried out in the absence of salt. Due to their poor retention on PGC-HPLC the analysis of single uronic acids has been optimised with high pH anion-exchange chromatography. Fluorescent labelled derivatives formed by reductive amination of neutral oligosaccharides with 2-aminobenzamide have been chromatographed on PGC-HPLC and by BioGel P4 gel filtration.
Davies
M J
MJ
Department of Biochemistry and Molecular Biology, University College London, UK.
Hounsell
E F
EF
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange; D005456:Fluorescent Dyes; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification*; D006108:Graphite; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification*; D011509:Proteoglycans / Q000737:chemistry / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D005456:Fluorescent Dyes; D006023:Glycoproteins; D009844:Oligosaccharides; D011509:Proteoglycans; D006108:Graphite