Preparation And Evaluation Of Open Tubular Electrochromatography Column With The BSA/RNA-PAMAM Dendrimers As Bonded Stationary Phases

Capillary electrochromatography (CEC) has been regarded as a very promising electrokinetic separation technique which combines the high efficiency of capillary zone electrophoresis (CZE) and the high selectivity of high-performance liquid chromatography (HPLC). OT-CEC has a great deal of advantages over packed capillary including ease of preparation, variety and availability of surface modification and immobilization chemistries, no bubble formation and higher compatibility with smaller internal diameter capillries. Moreover, the capillary of small diameters can be applied under the high voltage in CEC without remarkable Joule heating.Based on our previous research, the polyamidoamine star-burst dendrimers (PAMAM) were bonded to the interior wall of capillaries through alkylation reaction with 3-aminopropyltriethoxysilane, the Michael addition reation with methylarylate and amination reaction with athylenediaine.The BSA-PMAMA dendrimer was bonded to inner wall of capillary as staionary phases for open tubular capillary electrochromatography (OT-CEG). The obtained columns were characterized by labelling BSA with Fluorescein isothiocyanate (FITC). Under the optimized condition, the resolution of DL-Trp was gradually improved with the increase of PAMAM generation. By OT-CEC column modified with BSA-G3.0-PAMAM, the baseline separation of DL-Trp can be obtained. The separation of ovalbumin was studied by obtained CEC column.RNA ligand was coupled to terminal amido of OT-CEC column modified with different generation of PAMAM. Under the optimization condition, the lysozyme/ribonuclease A andβ-casein/α-lactalbumin were successfully separated by BSA-G3.0-PAMAM affinity electrochromatography column.