Improved capillary electrophoretic separation and mass spectrometric detection of oligosaccharides
Hyun Joo An1, Andreas H Franz, Carlito B Lebrilla
1Department of Chemistry, University of California, Davis, CA 95616, USA.
Abstract:
We have developed a CE method for the separation of structural isomers of oligosaccharides labeled with N-quaternized benzylamine. Oligosaccharides with reducing ends were derivatized with benzylamine by reductive amination followed by quaternization to yield a fixed cation label. The benzylamine-derivatized oligosaccharides were analyzed by CE-UV in ammonium acetate buffer and off-line matrix-assisted laser desorption ionization (MALDI) MS. The method was applied to a 1 nmol sample of a model oligosaccharide (LNDFH 1). From this sample a 38 fmol diluted standard was detected. The quaternization of benzylamine-labeled products significantly improved CE separation of neutral oligosaccharides along with several structural isomers. Two hexasaccharide isomers (LNDFH I and LNDFH II) were baseline resolved using an ammonium acetate buffer. This method was also applied successfully to the profiling of oligosaccharides released from the glycoprotein RNase B. The release of 6 pmol of glycans followed by workup showed the detection of all components, with one component corresponding to 100 fmol. Micropreparative collection of CE enabled successful off-line CE-MALDI-MS without additional sample clean up. This report provides a simple and rapid method to separate and analyze oligosaccharides.
Related Concept Videos
Oligosaccharide Assembly
Multiple sugar molecules that may or may...
Surface Tension, Capillary Action, and Viscosity
The various IMFs between identical molecules of a substance are examples of cohesive forces. The molecules within a liquid are surrounded by other molecules and are attracted equally in all directions by the cohesive forces within the liquid. However, the molecules on the surface of a liquid are attracted only by about one-half as many molecules. Because of the unbalanced molecular attractions on the surface molecules, liquids contract to form a shape that minimizes the number...
Capillary Beds
Capillaries connect arterioles, small branches of arteries, to venules,...
Capillary Exchange
Atomic Mass
Capillaries and Their Types


