Abstract
Rapid and simple achiral and chiral HPLC assays have been developed for the determination of verapamil and its metabolites in serum samples. Two achiral reversed-phase columns, Hisep C 18 (150×4.6 mm) and NovaPak C 18 (150×3.9 mm) were used for the simultaneous separation of all analyzed compounds. An α 1-AGP column (100×4.0 mm) was recommended for successful chiral separations of verapamil and its seven metabolites. All analyses were realised with fluorescence detection at λ ex=276 nm and λ em=310 nm. Limits of quantitation were in the range 1.0 to 5 ng/ml for all compounds. Both off-line SPE (SepPak C 18 cartridges) and the on-line SPE with a semipermeable surface SDS C 8 pre-column, (10×4.6 mm) were used for the clean-up and sample preconcentration. Extraction recoveries for all analyzed compounds were 87.7±5.8 to 92.7±4.0% for off-line SPE and 94.3±4.2 to 98.2±5.1% for on-line SPE. The complete assay could be applied for achiral and chiral monitoring verapamil and all its metabolites in serum samples.
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