Novel Support Material with Polymeric Ligand for Ion Exchange Chromatography: Synthesis, Characterization and Chromatographic Performance

Authors : Berna SARAÇOĞLU KAYA, Ali TUNCEL

Pages : 173-183

View : 22 | Download : 9

Publication Date : 2019-12-23

Article Type : Research Paper

Abstract :Within the scope of the study, polymeric ligands with weak cation exchange carboxyl insert ignore into journalissuearticles values(-COOH); groups formed on the surface, designed for High Performance Liquid Chromatography insert ignore into journalissuearticles values(HPLC);, monodisperse and porous polyinsert ignore into journalissuearticles values(2,3-dihydroxypropyl methacrylate-co-ethylene dimethacrylate);, polyinsert ignore into journalissuearticles values(DHPM- co-EDM); particles were tested in Ion Exchange Chromatography insert ignore into journalissuearticles values(IEC); mode for chromatographic separation of protein mixtures as a weak cation exchange column-material. The synthesis of the particles obtained for the design of the column support material, which is the first step of the study carried out in three stages, takes place in four different sections. Firstly, monodisperse polystyrene microgels to form the skeletal structure were synthesized by “Dispersion Polymerization”, then monodispersed-porous polyinsert ignore into journalissuearticles values(glycidyl methacrylate-co-ethyleneglycol dimethacrylate);, polyinsert ignore into journalissuearticles values(GMA-co-EDM); microspheres with epoxy groups were produced by using “ multi-stage microsuspension polymerization ”. The particles were hydrolyzed in acidic medium to form the surface in diol form and polyinsert ignore into journalissuearticles values(2,3-dihydroxypropyl methacrylate-co-ethylene dimethacrylate);, polyinsert ignore into journalissuearticles values(DHPM-co-EDM); particles were obtained which were suitable for chromatographic use. The particles obtained in the last step were used in the “Free Radical Grafting Polymerization” insert ignore into journalissuearticles values(FRGP); method for the production of polymeric ligand containing carboxyl group. Poly insert ignore into journalissuearticles values(DHPM-co-EDM); particles with –OH groups on the surface are silanated with 3-trimethoxysilylpropyl methacrylate insert ignore into journalissuearticles values(TMSPM); and derivatized with methacrylic acid insert ignore into journalissuearticles values(MAA); to form polyinsert ignore into journalissuearticles values(MAA); ligand on the surface of polyinsert ignore into journalissuearticles values(DHPM-c-EDM); particles were obtained for ion-exchange chromatography. The characterization and chromatographic performance of the particles were investigated in the second and third stages of the study. For particles synthesized by the FRGP method, the average-pore size was 33 nm and the specific-surface area was 21 m 2 /g. Theoretical plate height values ​​between 200-320 µm were obtained for weak cation exchange column. The peak resolution values ​​in the chromatogram were found to be above 1.4 for separation as short as 15 minutes. Protein recovery values ​​were found to be between 90-100% for all proteins. For the produced polyinsert ignore into journalissuearticles values(MAA);-grafted-polyinsert ignore into journalissuearticles values(DHPM-co-EDM); column, the BSS values ​​of the analytes from analysis to analysis and from day to day reproducibility tests were below 1%. These results showed that synthesized column packed materials can be successfully used in ion exchange chromatography.  
Keywords : Free Radical Grafting Polymerization FRGP, High Performance Liquid Chromatograpy HPLC, Ion Exchange Chromatography