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

The proteins cellulase and bovine serum albumin (BSA) have in this stu dy been cross-linked with glutaraldehyde to forma gel which has been u sed in capillary affinity gel electrophoresis (CAGE) to resolve enanti omeric pairs of beta-adrenergic blockers. Both proteins have earlier b een 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. Sin ce it was difficult to obtain a stable gel with cellulase alone, we co polymerized 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 capillar ies have an inner diameter of 75 mu m, a total length of 23,5 cm and a n 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 e lectrokinetically introduced and separated at a voltage of 3-3,5 kV. W ith this study we want to propose a new model based on copolymerizatio n of proteins, as chiral selectors, to create a gel which has the pote ntial to resolve different types of chiral compounds in capillary affi nity gel electrophoresis.