Abstract
Abstract An isocratic, rapid, sensitive and selective reversed phase high-performance liquid chromatographic (HPLC) assay with ultraviolet detection has been used to quantify the in vitro nimodipine metabolism in rat liver microsomes. (±) Nimodipine and its metabolites were separated on a C18 HPLC column maintained at 40°C using a mobile phase consisting of ammonium acetate 0.05M pH 6.6 and methanol (40:60 v/v). The within- and between-run coefficients of variation (CVs) were < 4.2 % in the concentration range of 0.5–50 μM. The limit of detection for nimodipine and ist metabolites was in the range of 30 - 80 nM. The formation of nimodipine metabolites may be described by a sigmoid Vmax model according to Hill equation corresponding to enzyme kinetics associated with positive allosteric effect. The assay was accurate, selective and may be used in studies investigating the interaction of drugs, and substances found in daily food e.g. flavonoids, with nimodipine metabolism.
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