Preparation And Application Of Novel Ion Exchange Chromatographic Stationary Phases

Unstirred in-situ polymerization method was employed to directly prepare poly(GMA-co-EDMA) monoliths within the confines of chromatographic column in the presence of porogens. Effects of key variables such as temperature, composition of the pore-forming solvent mixture, and content of cross-linking divinyl monomer on porous structure of monoliths were determined. Bimode micro porous and penetrability was investigated by mercury intrusion method and scanning electron micrograph. Weak anion exchange monoliths, anion exchange perfusive chromatographic columns and cation exchange perfusive chromatographic columns were achieved by the modification of epoxide groups of the monoliths, which reacted with diethylamine and iminodiacetic acid to afford l-N,N-diethylamino-2-hydroxypropyl and carboxyl group functionalities , respectively. Effects of some variables such as temperature, buffer concentration and pH on the binding capacity and breakthrough curves of the columns were examined. Dynamic binding capacity of Bovine Serum Albumin (BSA) and lysozyme on the anion and cation columns was 36.8mg/mL and 12.5mg/mL, respectively. Preliminary results confirmed the excellent transfer performance of the columns, thus, they were suitable for the quick separation of biopolymers. The columns were applied to separate a homologous series of proteins and oligonucleotides with very good results.