Enantioselective Determination of Oxprenolol in Human Plasma Using Dialysis Coupled on-Line to Reversed-Phase Chiral Liquid Chromatography

A fully automated method for the determination of the enantiomers of oxprenolol in human plasma was developed, involving dialysis through a cellulose acetate membrane, clean-up and enrichment of the dialysate on a short precolumn and subsequent chiral liquid chromatographic (LC) analysis. All sample handling operations were executed automatically by a sample processor equipped with a robotic arm (ASTED system). The trace enrichment column (TEC) was packed with octadecylsilica. After conditioning of the TEC with the LC mobile phase and pH 3.0 acetate buffer. After the enrichment step, the analyte was transferred by the LC mobile phase to the analytical column by means of a switching valve. The influence of different parameters of the dialysis process on the recovery of oxprenolol was first investigated using achiral LC conditions. The volume as well as the aspirating and dispensing flow rates of the acceptor solution were the main parameters studied. Oxprenolol was separated on a C18 stationary phase used for the enantioseparation of oxprenolol was a Chiralcel OD-R column which contained cellulose tris (3,5-dimethylphenylcarbamate) coated on silica as chiral selector. The corresponding mobile phase consisted of a mixture of pH 6.0 phosphate buffer containing NaClO4 at 0.45 M concentration and acetonitrile (70:30 v/v). UV detection was performed at 273 nm. The method developed was validated. Recoveries for each enantiomer of oxprenolol were about 80%. The method was found to be linear in the 50-2500 ng ml-1 concentration range (r2 = 0.999 for both enantiomers) and good results with respect to intra- and inter-day reproducibility as well as accuracy were obtained.