Liquid chromatographie separation of penicillamine enantiomers derivatized with OPA/2-ME on aβ-cyclodextrin bonded phase

Abstract

A high performance liquid Chromatographic method has been developed for the resolution of penicillamine enantiomers on a β-cyclodextrin (β-CD) column. Fluorogenic derivatives of the D,L-penicillamine were first formed by usingo-phthaldehyde/2-mercaptoethanol (OPA/2-ME) as derivatizating reagent. These chiral derivatives were then chromatographed on a commercially available chiral stationary phase of β-CD with 50/50 ethanol/1% triethylammonium acetate (pH 4.5) as a mobile phase, and detected fluorimetrically at 450 nm (λex = 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).