High-performance liquid chromatographic analysis of galactosamine, glucosamine, glucosaminitol, and galactosaminitol.
Both N-acetylgalactosamine and N-acetylglucosamine covalently link oligosaccharides to peptide in glycoproteins. In order to identify the N-acetylhexosamine involved in this linkage, the corresponding hexosaminitol generated by alkaline borohydride treatment must be determined. An HPLC method modified from the Waters PICO-TAG amino acid analysis procedure is described. Phenylisothiocarbamyl derivatives of galactosamine, glucosamine, glucosaminitol, galactosaminitol, and the internal standard, p-aminophenyl-beta-D-galactoside, are eluted from the Waters PICO-TAG column at 3.9, 4.3, 6.9, 8.1, and 10.1 min, respectively. The standard curves for the hexosamines and hexosaminitols are linear between 1 and 75 nmol. In addition to p-aminophenyl-beta-D-galactoside, several synthetic hexosamines and hexosaminitols can be employed as internal standard. These include 3-allosamine, 3-glucosamine, allosamine, 3-allosaminitol, mannosaminitol, and allosaminitol, which are eluted at 3.1, 3.4, 4.8, 6.4, 7.4, and 7.8 min, respectively. The analysis time is 15 min but can be shortened to 10 min if only hexosamines are to be analyzed and either 3-glucosamine or 3-allosamine is used as the internal standard. This rapid method is superior to previous methods for the analysis of hexosamines in glycoconjugates and hexosaminitols generated from glycoconjugates following alkaline borohydride treatment.
Department of Pediatrics, University of North Carolina, Chapel Hill 27514.
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Cheng
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005688:Galactosamine / Q000032:analysis; D005944:Glucosamine / Q000031:analogs & derivatives / Q000032:analysis; D006595:Hexosamines / Q000032:analysis*; D006801:Humans; D009077:Mucins / Q000032:analysis
D006595:Hexosamines; D009077:Mucins; C020862:2-amino-2-deoxyglucitol; D005688:Galactosamine; D005944:Glucosamine