Reverse phase HPLC fractionation of the oligosaccharide alditols isolated from an I-active ovarian cyst mucin glycoprotein

We have used reverse phase high pressure liquid chromatography (HPLC) to separate the reduced oligosaccharides produced by alkaline borohydride degradation of ovarian cyst blood group substances. From a single cyst, six oligosaccharides, ranging from two to seven residues in length, have been isolated by preparative HPLC on C-18 stationary phases using water for elution. The purity of the products and their structures were determined by high field proton NMR spectroscopy in conjunction with exo-and endoglycosidase digestion. All the chains isolated terminated inN-acetylgalactosaminitol which was substituted at the 3-postion by galactose and in some cases at the 6-postion byN-acetylglucosamine. The largest identified oligosaccharide was a heptasaccharide alditol containing a single -linked fucose in a Lewis blood group structure (Le^a).