Analysis of sialic acid-containing mucin oligosaccharides from porcine small intestine by high-temperature gas chromatography-mass spectrometry of their dimethylamides.
Acetylation of sialic acid-containing oligosaccharides lactonises the sialic acid residue quantitatively for all oligosaccharides studied except for 6'-sialyl-lactose. The modified, unsulphated, sialylated and sulphated oligosaccharides can then be fractionated by anion-exchange chromatography. Ammonolysis of the lactones followed by methylation yielded the dimethylamides, which are amenable to g.l.c.-m.s. and give intense and informative mass spectra. This approach has been used to characterise the sialic acid-containing O-linked oligosaccharides obtained from the mucin glycopeptides of the small intestine of the pig. At least 28 structures were found, having NeuAc or NeuGc 6-linked to the HexNAc attached to the peptide core or to a Hex 3-linked to HexNAc. Four different disialylated oligosaccharides were found having NeuAc or NeuGc on the Hex residue 3-linked to HexNAc and 6-linked to HexNAc.
Hansson
G C
GC
Department of Medical Biochemistry, University of Göteborg, Sweden.
Bouhours
J F
JF
Karlsson
H
H
Carlstedt
I
I
D000818:Animals; D002240:Carbohydrate Sequence; D004123:Dimethylamines / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry; D007421:Intestine, Small / Q000737:chemistry; D008969:Molecular Sequence Data; D009077:Mucins / Q000737:chemistry*; D019158:N-Acetylneuraminic Acid; D009844:Oligosaccharides / Q000737:chemistry*; D012794:Sialic Acids / Q000737:chemistry*; D013552:Swine
D004123:Dimethylamines; D009077:Mucins; D009844:Oligosaccharides; D012794:Sialic Acids; C034516:dimethylamine; D019158:N-Acetylneuraminic Acid