Preparation of sub‐micron skeletal monoliths with high capacity for liquid chromatography

A novel kind of poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate)-based monolithic column was developed for LC by directing supramolecular self-assembly of high internal phase emulsion. Mercury intrusion porosimetry characterization and scanning electron microscope pictures showed that these monoliths presented micrometer-sized throughpores, unique sub-micron skeletons and relatively large specific surface area. Additionally, porosity of monoliths could be adjusted while skeletons remained in the size range of 100.0–1000.0 nm. The new monoliths demonstrated not only better column efficiency, but also larger binding capacity. Dynamic binding capacity for protein (BSA) was evaluated to be 42.5 mg/mL, above two times higher than that of the general monoliths (19.1 mg/mL) and higher than that of commercial “Convective Interaction Media” monolithic columns (30.0 mg/mL). Moreover, their chromatographic behaviors were also evaluated in detail by chemical stability and swelling characterization of the monolithic column. Separation of proteins mixture (cytochrome c, myoglobin, ribonuclease A, lysozyme and BSA) on the monolith was achieved within 4 min at velocity of 1440.0 cm/h. Those unique properties made the novel monolithic column a promising alternative to commercially available monolithic supports in LC applications.