SYNTHESIS AND CHARACTERIZATION OF SILICA-IMMOBILIZED SERUM-ALBUMIN STATIONARY PHASES FOR HPLC

Chiral HPLC stationary phases based on immobilized biopolymers (i.e., proteins, enzymes, antibodies etc.) are of growing importance for the separation and purification of enantiomeric compounds, especially in t he areas of biological, pharmaceutical, and clinical chemistry. In the work described in this paper, several silica-immobilized serum albumi n phases were synthesized and characterized by a variety of techniques . Low-temperature nitrogen adsorption isotherms were measured and empl oyed to evaluate the surface characteristics of the stationary phases as a function of both the immobilization matrix and the surface modifi cation chemistry. Elemental analysis and high resolution thermogravime try were used to evaluate the surface coverages at each stage of modif ication. Based on these results, surface parameters such as the specif ic surface area, pore size distribution, bonding density, etc. were ca lculated, as well as the changes of these parameters as a function of the modification chemistry were analyzed. Different silicas of varying mean pore diameter from 30 nm to 400 mm were used.