Size exclusion chromatography of polysaccharides with reverse phase liquid chromatography

This work describes the use of a reverse phase (RP) column for size-based separation of hydrophilic polysaccharides. The entire separation window (before and after void volume) is used to provide unique separation of polysaccharides from other components in a glycol-conjugate vaccine or complex media of fermentation broth. The technique has also been applied to the separation of polysaccharides of different sizes. The effect of chromatographic parameters including type of packing material (CN, C8 and C18), pore size (80 and 300A) of stationary phase, concentration of organic solvent on the separation was investigated. In addition, characterization of size-based separation was evaluated by coupling of RP column with multi-angle laser light scattering (MALLS) detector. (C) 2013 Elsevier B.V. All rights reserved.