Stereochemical resolution of racemates, in HPLC, using a chiral stationary phase based upon immobilized α-chymotrypsin.: I.: Structural chiral separations

alpha-Chymotrypsin was immobilized on an epoxide derivatized silica gel by an in situ immobilization process. Several racemates were resolved by a structural recognition mechanism. The immobilization process and the stability of this alpha-chymotrypsin stationary phase have been studied. The mobile phase parameters including the ionic strength, pH and the effects of organic modifiers have been also investigated. The retention, efficiency and stereoselectivity of the solutes appear to be related to their molecular structure, hydrophobicity and electrostatic interactions. These relationships determine the recognition mechanism and the position of each enantiomer in the active site.