[The determination of propranolol enantiomers in rat plasma by high-performance liquid chromatography with fluorescence detecion; development and validation.]
De bepaling van propranolol enantiomeren in ratteplasma met behulp van hoge-druk vloeistofchromatografie en fluorescentie detectie; ontwikkeling en validatie
Stolker AAM , Bode W , Toet AE , Wemer J , Ginkel LA van
RIVM Report 318601001
This report describes a method for the determination of the enantiomers of the beta-blocker propranolol. Propranolol is isolated by solid phase extraction. The propranolol enantiomers are derivatized to diastereomeric compounds by reaction with the chiral reagent (-)-menthylchloroformate. The diastereoisomers are separated by reversed-phase high-performance liquid chromatography (HPLC) on an octadecylcolumn, using methanol-water (875/125 ; v/v) as eluent. The diastereoisomers are detected by fluorescence detection. Standard curves for both R-(+)-propranolol and S-(-)-propranolol are linear in the 0,050-1,50 mg/l range. The limit of detection is 0,010 mg/l for each enantiomer. The limit of determination is 0,050 mg/l for each enantiomer based on a 50 mul plasma. The extraction recoveries are between 95-100%. Within-laboratory reproducibility (n=20) is 5,5% for S-(-)-propranolol and 5,9% for R-(+)-propranolol at 0,352 mg/l. The method has been used for pharmacokinetic studies of propranolol enantiomers in rats.