Capillary electrophoresis of N-acetylneuraminic acid polymers and hyaluronic acid:: Correlation between migration order reversal and biological functions

High-resolution analysis of polymers of N-acetylneuraminic acid and hyaluronic acid was performed by capillary electrophoresis in a buffer containing a neutral polymer. Both polysaccharides having more than 100 monosaccharide residues were well separated into their molecular species by capillary electrophoresis using a combination of a chemically modified capillary and a buffer containing poly(ethylene glycol) as an additive. During optimization of the separation conditions, small oligomers of both polysaccharides were observed to migrate in the reverse order of their molecular masses on the electropherograms. However, oligomers larger than pentamer and decamer for N-acetylneuraminic acid polymers and hyaluronic acid, respectively, migrated in the order of their molecular masses. We propose that these unusual migration patterns are closely related to the stereochemical structures and the oligomer migrating the fastest is the minimum unit that forms the regular three-dimensional structure required for the: biological function.