Key WordsColumn liquid chromatography c~-Chymotrypsin immobilized on silica In situ immobilization process Structural chiral separations Summary ct-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 ct-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.

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