PROTEIN-BASED CAPILLARY AFFINITY GEL-ELECTROPHORESIS FOR CHIRAL SEPARATION OF BETA-ADRENERGIC BLOCKERS

The proteins cellulase and bovine serum albumin (BSA) have in this study been cross-linked with glutaraldehyde to forma gel which has been used in capillary affinity gel electrophoresis (CAGE) to resolve enantiomeric pairs of beta-adrenergic blockers. Both proteins have earlier been used as chiral selectors, especially in HPLC. We have utilized the major quantitatively cellulase, cellobiohydrolase I (CBH I) produced by the fungus Trichoderma reesei, in CAGE to separate enantiomers. Since it was difficult to obtain a stable gel with cellulase alone, we copolymerized it with BSA. With this cellulase/BSA gel we could resolve the optical isomers of the beta-blocking drugs, rac-propranolol, rac- metoprolol, rac-pindolol and partially rac-atenolol. The used capillaries have an inner diameter of 75 mu m, a total length of 23,5 cm and an effective length of 15,5 cm filled with gel to the detection window. The buffer used for separation was, 50 mM potassium phosphate buffer at pH 6.8 with 1% 2-propanol added as organic modifier. Samples were electrokinetically introduced and separated at a voltage of 3-3,5 kV. With this study we want to propose a new model based on copolymerization of proteins, as chiral selectors, to create a gel which has the potential to resolve different types of chiral compounds in capillary affinity gel electrophoresis.