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We applied multiple relayed COSY and 2D homonuclear Hartman-Hahn spectroscopy to globoside, a glycolipid purified from human red blood cells. The subspectra corresponding to individual sugar components were extracted even from overlapping proton resonances by taking the cross sections of 2D spectra parallel to the F 2axis at anomeric proton resonances, so that unambiguous assignments of sugar proton resonances were accomplished.

We applied ID homonuclear Hartmann-Hahn (1D-HOHAHA) and difference NOE experiments to determine the chemical structure of Forssman's antigen, a glycolipid purified from sheep red blood cells. The subspectra corresponding to the individual sugar components were extracted from overlapping proton resonances by selective excitation of the anomeric proton resonances, so that unambiguous assignments of the sugar proton resonances were accomplished. Then, difference NOE experiments were performed to determine the linkage of the sugar units. The present procedure was found to be useful for the structure determination of glycoconjugates and also reduces the amount of samples and machine time.