Probing the microheterogeneity of O-specific chains from Yersinia ruckeri using capillary zone electrophoresis/electrospray mass spectrometry.

Abstract

The analysis of underivatized oligosaccharides arising from mild acid hydrolysis of endotoxins from Yersinia ruckeri serotype O2 was achieved using on-line capillary zone electrophoresis-electrospray mass spectrometry (CZE-ESMS). This technique provided unparalleled resolution of the different glycans obtained from purified fractions of the native endotoxins or from hydrolysis of lipopolysaccharides from Y. ruckeri. Electrophoretic conditions enabling the separation of anionic and cationic analytes were developed to determine possible sites of heterogeneity on either the core or the O-chain glycans. Structural characterization of underivatized oligosaccharides identified in the ion electropherograms was achieved using tandem mass spectrometry under low-collision energy conditions.