LIQUID-CHROMATOGRAPHIC SEPARATION OF PENICILLAMINE ENANTIOMERS DERIVATIZED WITH OPA/2-ME ON A BETA-CYCLODEXTRIN BONDED PHASE

A high performance liquid chromatographic method has been developed for the resolution of penicillamine enantiomers on a beta-cyclodextrin (beta-CD) column. Fluorogenic derivatives of the D,L-penicillamine were first formed by using o-phthaldehyde/2-mercaptoethanol (OPA/2-ME) as derivatizating reagent. These chiral derivatives were then chromatographed on a commercially available chiral stationary phase of beta-CD with 50/50 ethanol/1% triethylammonium acetate (pH 4.5) as a mobile phase, and detected fluorimetrically at 450 nm (lambda(ex) = 355 nm). After careful optimization of the classical chromatographic parameters good resolution was achieved between the D and L enantiomers. The method proposed here is simple and rapid and can detect the presence of 0.5% of the L-enantiomer in D-penicillamine pharmaceutical preparations (tablets).