DEVELOPMENT OF A FLAVOPROTEIN COLUMN FOR CHIRAL SEPARATION BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A new chiral stationary phase using flavoprotein, a glycoprotein present in chicken egg-white, was developed for high-performance liquid chromatography. This column could achieve baseline separations of acidic (ketoprofen), weakly acidic (warfarin) and neutral (benzoin) compounds. In the flavoprotein-conjugated silica gel column, the capacity factor and enantioselectivity for several model drugs were greatly influenced by the pH and the concentration of organic solvents and salts in the mobile phase, and the optimum conditions for chiral separation varied from compound to compound. However, this column shows good stability to pH variation and to organic solvents, and it should be applicable for the chiral separation of many compounds in the reversed-phase mode. This column can directly separate optical isomers with an aqueous mobile phase, so it should be very useful in the fields of pharmacokinetics and clinical chemistry.