Enantioselective determination of felodipine in human plasma by chiral normal-phase liquid chromatography and electrospray ionisation mass spectrometry.

ABSTRACT

An analytical method was developed for the determination of the enantiomers of felodipine, a dihydropyridine-type calcium antagonist, in human blood plasma. Felodipine was extracted from plasma using toluene as extraction solvent. The enantiomers were separated on a cellulose tris(4-methyl benzoate) stationary phase (Chiralcel OJ-R) using 2-propanol-iso-hexane (1189) as mobile phase. Post-column addition of ammonium acetate in ethanol-water (955) allowed sensitive detection of the ammonium adduct by electrospray ionisation and selected reaction monitoring. Deuterated felodipine racemate was used as internal standard. Within-run repeatability was determined and a coefficient of variation below 2% was achieved at 22 nmol/l and below 10% at 0.27 nmol/l. Between-day precision was evaluated and a coefficient of variation of 3.6% at 4 nmol/l plasma was obtained. Limit of quantification (LOQ) was set at 0.25 nmol/l (0.10 microg/l). The method proved adequate for pharmacokinetic studies of R- and S-felodipine after oral administration of therapeutic doses of felodipine.