The determination of the structure of blood group oligosaccharides from fully assigned 1H-n.m.r. spectra for solutions in non-aqueous solvents.

The fully assigned 1H-n.m.r. spectra of a blood group A tetrasaccharide and of a blood group H hexasaccharide in dimethyl sulfoxide and in pyridine by use of two-dimensional COSY and homonuclear Hartmann-Hann coherence transfer methods are reported. The 1H-n.m.r. spectra of both of these compounds in deuterium oxide had been previously assigned. Since the relative proton chemical shifts in the three solvents are quite different, resonances which overlap or are strongly coupled for one solvent may be well resolved for another, thus providing an extension of the method of complete proton assignments for determination of the structure of complex oligosaccharides. Although the rotational correlation times (tau c) of these oligosaccharides are similar to the reciprocal of the spectrometer frequency, either negative or positive n.O.e. values were measurable for both oligosaccharides in all three solvents in one-dimensional difference spectroscopy by taking advantage of the dependence of tau c on the solvent viscosity and, thus, on sample temperature. Whereas n.O.e. depend strongly on temperature and solvent viscosity, the ratios of the effects between protons on the same pyranoside ring and those on different rings were observed to be similar, suggesting that the oligosaccharide conformations are not strongly dependent on solvent or temperature.