Determination of Verapamil and Norverapamil in Serum by High-Performance Liquid Chromatography

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Abstract
A simple, rapid, sensitive and specific high-performance liquid chromatographic method is developed for the determination of verapamil and its active metabolite norverapamil in human serum. The sample (100 μl) is vortex-mixed with methanol (200 μl) and after centrifugation 100 μl of the supernatent is used for the analysis of a C8 reverse-phase column (10 cm × 5 mm) with methanol - 0.1 M ammonium acetate (55:45 v/v) as the mobile phase. The compounds are detected fluorimetrically with a detection limit of 5 μ/l using an excitation wavelength of 278 nm and an emission wavelength of 320 nm. The method is applicable to pharmacokinetic studies and to therapeutic monitoring of the drug and its active metabolite.