Preparation Of A Silica-microspheres-encapsulated/Hydrophilic Strong Cation-exchange Monolithic Column

Micro-column liquid chromatography is a high efficient separation technique, it hasgrown in importance for proteomics research due to its high resolving power, wildlyapplication, excellent reproducibility. Nontheless, current one-dimensional liquidchromatographic techniques aren’t yet satisfactary for the separation of peptide and proteinmixtures because of its quite higher complexities. Therefore, an orthogonal two-dimensional(2-D) or multi-dimensional separation system is required since it can provide a higher peakcapacity. Hydrophilic strong cation-exchange chromatography is an important dimension for2-D or multi-dimensional liquid chromatography because it can provide a suitableenvironment for biologically active compounds and is highly orthogonal to other separationmodes such as reversed phase chromatography.In the present study, a silica-microspheres-encapsulated/hydrophilic strongcation-exchange monolithic column was prepared by direct in-situ encapsulation of silicamicrospheres with organic polymers of3-sulfopropyl methacrylate potassium salt andpoly(ethyleneglycol) diacrylate in a530μm i.d. fused silica capillary. The composition ofporogen, optimal total concentration of monomers and time of polymerization have beendetermined by comparing the permeability, scanning electron micrographs, curves ofbackpressure vs flow-rate of mobile phase and curves of Van Deemter obtained with thecolumns prepared under different conditions. The prepared monolithic capillary column wasevaluated in terms of pore structure, permeability and anti-swelling.