Assay of keratan sulfate as anion-exchanger bound hexose.
An assay for keratan sulfate in papain digests of human intervertebral disc and other tissues has been developed. The digest is applied to the acetate form of a tertiary amine acrylic anion-exchange resin, the oligosaccharide hexose is removed by washing the resin with 0.2 M sodium acetate buffer pH 5.0, then the keratan sulfate is eluted quantitatively with 1.0 M pyridinium sulfate pH 2.5 and assayed for hexose by the anthrone reaction. The keratan sulfate content of human intervertebral disc tissues ranged from 7 to 78 mumole galactose equivalents/g fresh weight; the root mean square error was 2 mumole/g; 10-25 mg of tissue were required. The separation of oligosaccharides from keratan sulfate was confirmed by gel permeation chromatography, sugar composition, ester sulfate analysis, and nuclear magnetic resonance.
Bebault
G M
GM
Department of Pathology, Faculty of Medicine, University of British Columbia, Vancouver, Canada.
Pearce
R H
RH
D000085:Acetates; D019342:Acetic Acid; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006601:Hexoses / Q000032:analysis*; D006801:Humans; D007202:Indicators and Reagents; D007403:Intervertebral Disc / Q000737:chemistry; D007632:Keratan Sulfate / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D009844:Oligosaccharides / Q000032:analysis; D010206:Papain; D011726:Pyridinium Compounds
D000085:Acetates; D006601:Hexoses; D007202:Indicators and Reagents; D009844:Oligosaccharides; D011726:Pyridinium Compounds; D007632:Keratan Sulfate; D010206:Papain; D019342:Acetic Acid