A high-performance liquid chromatographic (HPLC) method for the measurement of amiodarone (AM) and its metabolite(s) in serum and tissues was developed. The method uses a 5-micron silica column, methanol containing 0.02% perchloric acid at pH 4 as the mobile phase, and ultraviolet detection at 240 nm. The standard curves for AM and desethylamiodarone (DAM) were linear for serum (range 0.025-6.0 microgram/ml) and tissues (range 0.1-0.5 micrograms for 10-25 mg wet weight). There was a significant decrease as a function of time in AM and DAM concentrations in patients' sera left at ambient temperature in the presence of light. This HPLC method was applied to studies on serum AM elimination kinetics in patients and on tissue uptake during chronic AM administration to rabbits. The elimination half-life (5.8 h) of AM after a 5 mg/kg intravenous dose to a patient was similar to that after acute oral doses. AM, being lipophilic, accumulated maximally in the fat tissue (56 micrograms/g wet weight), followed by lung and liver in rabbits injected with AM for six weeks. The latter two tissues also contained nearly equal quantities of DAM. The high concentrations of AM and DAM in the liver and lungs may be related to the hepatic toxicity and pulmonary fibrosis associated with chronic AM therapy. Two new metabolites were found in the lung and bile of AM-treated rabbits, but these have not yet been identified.