Preparation Of Series Sulfated Oligosaccharides And Their Structural And Sequential Analysis

Carrageenan, a kind of sulfated-galactan, which extracted from marine red alga, and heparin, which extracted from animal tissue, a kind of acidic mucopolysaccharide, have many activities, such as, anticoagulant, antiviral and anticancer activities etc. All the two kinds of sulfated polysaccharides regarded as model biomolecular used in the carbohydrate-based drug research and development. In this paper, the kappa-carrageenan was hydrolyzed by dilute HC1, and heparins from bovine lung and porcine mucous were hydrolyzed by heparmase I, and the oligosaccharides mixture was separated by Low-pressure gel permeation chromatography (LPGPC) and strong-anion-exchange-HPLC. Twenty-five kappa-carrageenan derived oligosaccharides and fifteen kinds of heparin-derived unsaturated oligosaccharides were acquired. All the structure and sequence of these oligosaccharides were confirmed by 'H-NMR, 13C-NMR, 1H 'H-COSY, *H 13C-HMQC, HMBC, and also FAB/MS and ESIMS techniques. The oligosaccharides prepared from kappa-carrageenan were beta-carribiose, and different sulfate content carrageenan-derived trisaccharide, pentasaccharides, heptasaccharides, nonasaccharides, undecasaccharides, tridecasaccharides, pentadecasaccharides, heptasaccharides, nonadecasasaccharides, and henicosasaccharides. The heparin-derived oligosaccharides prepared by heparinase I from bovine and porcine were unsaturated fully sulfated and under sulfated heparin-derived disaccharide, tetrasaccharides, hexasaccharides, octasaccharides, decasaccharide,dodecasaccharides, tetradecasaccharide, hexadecasaccharides, octadecasaccharide, and eicosasaccharides. Except for the beta-carribiose, all other kappa-carrageenan derived oligosaccharides were new compounds.Based on the structure and sequential analysis of kappa-carrageenan oligosaccharide, the mild acid hydrolysis proved only occurred at a -1, 3-glycoside linkage and form odd number oligosaccharides which reducing end is galactose-4-sulfate and non-reducing end was galactose or galactose-4-sulfate. Owning to the ESIMS and FACE analysis of kappa-carrageenan derived oligosaccharide, the original carrageenan should be linear sulfated galactan with no branch. All these structural determined sulfated oligosaccharides offered basic data and will be further used, from the molecular lever, in the structure -activity study.