Stereochemical resolution of racemates, in HPLC, using a chiral stationaryphase based upon immobilized alpha-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 str uctural 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 organi c modifiers have been also investigated. The retention, efficiency and ster eoselectivity of the solutes appear to be related to their molecular struct ure, hydrophobicity and electrostatic interactions. These relationships det ermine the recognition mechanism and the position of each enantiomer in the active site.