STRATEGY FOR THE INVESTIGATION OF O-LINKED OLIGOSACCHARIDES FROM MUCINS BASED ON THE SEPARATION INTO NEUTRAL, SIALIC-ACID-CONTAINING AND SULFATE-CONTAINING SPECIES

A method for the separation of O-linked oligosaccharides into neutral, sialic acid-containing and sulfated species was applied to oligosacch arides released by alkaline borohydride from mucin glycopeptides from porcine small intestine. The released mixture of reduced oligosacchari des was applied to an anion exchange column, and the neutral oligosacc harides were collected as the unretarded fraction. A mixture of dimeth yl sulfoxide and iodomethane was passed through the column to convert the sialic acid-containing oligosaccharides into methyl esters that we re eluted and converted to methyl amides by methyl amine. Finally the sulfated oligosaccharide fraction was eluted with salt, The neutral an d the derivatized sialic acid-containing oligosaccharides were analyse d by gas chromatography-mass spectrometry a after permethylation and t he sulfated oligosaccharide fraction was analysed by high performance anion exchange chromatography.