Structural elucidation of zwitterionic sugar cores from glycosphingolipids by nanoelectrospray ionization-ion-trap mass spectrometry

The use of electrospray ionization (ESI)-ion-trap mass spectrometry (ITMS) for analysis of zwitterionic, glycolipid-derived sugar cores of glycosphingolipids is described. The capability of the method to perform multiple steps of fragmentation (MSn) allows structural characterization of these compounds. No derivatization of the released oligosaccharides is necessary when using nano-ESI with sample solution flow rates of about 30 nL/min. Investigations of positive as well as negative ions in fragmentation experiments up to MS4 permit determination of the sequence of sugar units, their linkage positions, and the exact location of the substituents phosphocholine and phosphoethanolamine, In the case of phosphocholine, chemical cleavage of this substituent was necessary to obtain all the linkage information. Approximately 150-250 ng of sample was needed for each analysis. (C) 2000 Academic Press.