Retention Characteristics of Protein-Based Chiral HPLC Columns

Abstract

The retentivity and enantioselectivity of chiral HPLC columns containing immobilized stereoselective proteins as the stationary phase were investigated by varying the parameters of sample size, column temperature, mobile phase pH, mobile phase organic modifier and inorganic buffer concentration. Columns containing bovine serum albumin, ovomucoid, and alpha(l)-acid glycoprotein were evaluated and retention characteristics were compared by separating acidic and basic test compounds. Enantioselectivity of each of these protein-based chiral columns is affected somewhat differently by the chromatographic parameters; chiral separations were optimized by adjustment of the appropriate parameters.