Chromatographic Performance of Weak Cation Exchanger Synthesized by a Simple Method

The column materials in the form of monodisperse and porous particles were synthesized with regard to test their column performances in Ion Exchange Chromatography (IEC) mode. Particles in the form of poly(glycidil methacrylate-coethyleneglycol dimethacrylate) poly(GMA-co-EDM) copolymer used for starting materials in the synthesis, were maintained by “multi-stage microsuspension polymerization”. Particle characterization was assessed by Scanning Electron Microscopy (SEM), BET surface area and porosity measurement system and FTIR spectroscopy. Weak cation-exchange carboxyl (- COOH) group on monodisperse and porous poly(DHPM-co-EDM) particles, used for ion-exchange chromatography, was synthesized by a simple method. The particles were hydrolyzed with trifloroacetic acid (TFA) in order to generate carboxyl groups on their surfaces. By use of this approach, a new column type having small ion-exchange ligands, are maintained. The investigation of column performances concluded that weak cation-exchanged column, synthesized by acidic hydrolysis (200- 350 µm), showed low and stable theoretical plate heights and high peak resolution values compared to literature knowledge. Protein recovery data in ion-exchange column, which is synthesized by acidic hydrolysis (AH) (94-99%) polymerization technique, was determined as similar as quantitative values for all proteins. Reproducibility tests of analytes of poly(MAA) column was assessed “during analysis and day by day”. The BSS values in column, synthesized by acidic hydrolysis, were determined as 2.51% and 1.75%, respectively. Results showed that synthesized column material can be successfully used in ion-exchange chromatography