In-source fragmentation and analysis of polysaccharides by capillary electrophoresis/mass spectrometry.
In order to develop a robust and easy-to-use technique for characterization of bacterial polysaccharides, a pseudo-hydrolysis strategy was investigated. Based on in-source collision-induced dissociation, polysaccharide molecular ions were fragmented within the orifice-skimmer region of an electrospray ionization (ESI) mass spectrometer. The fragment ions thus generated were then analyzed similarly to the conventional ESI mass spectrometry approach. MS/MS scanning was applied to obtain product-ion spectra of the primary fragments for sequencing. To further improve the sensitivity and separation of polysaccharides from other components in the samples, a pressure-assisted capillary electrophoresis/mass spectrometry (CE/MS) system was employed. Using bacterial polysaccharides as model compounds, the mass spectra obtained for polysaccharide repeating units generated through chemical hydrolysis and in-source fragmentation were directly compared, both in positive and negative ion modes. With the additional separation of impurities provided by CE, the success of this technique has been demonstrated for structural analysis of O-chain polysaccharides (O-PS) and capsular polysaccharides (CPS). In-source fragmentation was applied to promote the formation of structurally relevant repeating units of heterogeneous CPS that would remain undetected using conventional ESI conditions. This approach was proven to be particularly useful for probing the subtle structural differences in monosaccharide composition and functionalities arising across bacterial serotypes.
Li
Jianjun
J
Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario. Jianjun.Li@nrc-cnrc.gc.ca
Wang
Zhan
Z
Altman
Eleonora
E
D000333:Aeromonas / Q000737:chemistry; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D019075:Electrophoresis, Capillary; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011135:Polysaccharides, Bacterial / Q000032:analysis* / Q000737:chemistry*
D011135:Polysaccharides, Bacterial