SEPARATION OF ENANTIOMERS USING CELLULASE (CBH-I) SILICA AS A CHIRAL STATIONARY PHASE

A new chiral stationary phase for high-performance liquid chromatography based on a glycoprotein (cellulase, CBH I) isolated from a culture filtrate of a fungus, Trichoderma reesei, was prepared. Chiral acidic and basic drugs were resolved into their enantiomers on this phase. Compared with other similar chiral phases, high enantioselectivity was obtained for beta-blocking agents despite low capacity factors. As much as 200 nmol of propranolol in a single injection could be separated into its enantiomers on an analytical column (250 x 5.0 mm I.D.) without loss of resolution. No significant decrease in enantioselectivity was observed after daily use of the phase during a period of 4 months with varying mobile phase compositions. The retention and enantioselectivity of amines increased with increasing pH of the mobile phase, whereas the opposite effect was observed for acids. Addition of organic solvents to the mobile phase both decreased the retention and increased the enantioselectivity for the analytes.