Determination of (R)- and (S)-propranolol in plasma by high-performance liquid chromatography using N-benzoxycarbonylglycyl-L-proline as chiral selector in the mobile phase.

A normal-phase chromatographic method for the determination of (R)- and (S)-propranolol in plasma is described. The chiral separation is performed by adding an optically active complexing agent, N-benzoxycarbonylglycyl-L-proline, to the mobile phase (dichloromethane). The solid phase is LiChrosorb DIOL. After adjustment of the pH of the plasma, the propranolol enantiomers are extracted into hexane-dichloromethane-n-butanol (72:18:10). The organic phase is evaporated and the residue dissolved in the mobile phase before injection on to the column. Quantifications are performed by using internal standardization, giving a precision of better than 2% (coefficient of variation). The method employs 1-ml plasma samples and has linear calibration graphs (r = 0.999) over the concentration range studied, 9.2-288 nmol/l. injections of sample solutions with a composition different from that of the mobile phase gave system peaks that might affect the shape of the solute peaks. Several possibilities for avoiding these disturbing system peaks in the chromatogram by changing the mobile phase composition are discussed.