Novel highly hydrophilic zwitterionic monolithic column for hydrophilic interaction chromatography

A novel zwitterionic hydrophilic porous poly(SPV-co-MBA) monolithic column was prepared by thermal co-polymerisation of 1-(3-sulphopropyl)-4-vinylpyridinium-betaine (4-SPV) and N,N'-methylenebisacrylamide (MBA). An HILIC/RP dual separation mechanism was observed on this optimised poly(SPV-co-MBA) monolithic column and the composition of the mobile phase corresponding to the transition from the HILIC to the RP mode was around 30% ACN in water. Higher hydrophilicity was achieved on this novel monolithic column compared to the poly(N,N-dimethyl-N-methacryloxyethyl-N-(3-sulphopropyl)ammonium betaine-co-ethylene dimethacrylate) monolithic column. Permeability studies showed slight swelling and/or shrinking with mobile phases of different polarity. As might be anticipated, a weak electrostatic interaction for charged analytes was also observed by studying the influence of mobile phase pH and salt concentration on their retention on the poly(SPV-co-MBA) monolithic column. The final optimised poly(SPV-co-MBA) monolith showed comparable selectivities to commercial ZIC-pHILIC phases for polar test analytes. Fast separation of five pyrimidines and purines was achieved in less than 1 min due to the high permeability of the monolithic column. Additionally, baseline separation of nine benzoic acid derivatives was also observed using either a pH or ACN gradient.