An HPLC Method for the Determination of Diltiazem and Three of its Metabolites in Serum

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Abstract
A high performance liquid chromatographic method was developed for the determination of diltiazem and three of its metabolites in serum. Serum samples are extracted with methyl-tert.-butyl ether followed by back-extraction into 0.05N sulfuric acid. Doxepin hydrochloride was used as internal standard. A cyanopropylsilane column in the reverse-phase mode was utilized with a mobile phase of acetonitrile-phosphate buffer-triethylamine (pH 3.5). Using UV detection at 237nm, the lower limit of sensitivity was 1.5–3.0 ng/ml. Recovery and reproducibility results and possible interferences from other compounds are presented and discussed. The assay procedure was applied to a chronic oral dosing study in humans to assess the disposition of diltiazem and its primary metabolites.

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