Rapid high-performance liquid chromatographic analysis of amiodarone and N-desethyl-amiodarone in serum

Abstract

A rapid and reliable high-performance liquid chromatographic (HPLC) assay has been developed for the measurement of the antiarrhythmic drug, amiodarone (A), and its metabolite, N-desethyl-amiodarone (NDA), in serum. The procedure involves addition of absolute ethanol to serum, previously adjusted to pH 6, followed by centrifugation. The supernatant is analyzed by reverse-phase HPLC with ultraviolet–visible detection. There is no intereference from endogenous substances or other commonly used antiarrhythmic agents. The mean overall accuracy of the HPLC assay is 96.9 ± 9.1 (SD)% (n = 37) for A over the concentration range 0.10–10.00 μg/mL, and 99.8 ± 4.8 (SD)% (n = 32) for NDA over the concentration range 0.10–7.50 μg/mL. The within-day coefficient of variation is less than 7% for A and NDA. The lower limit of quantitative sensitivity is 0.10 μg/mL for A and NDA, and the lower limit of qualitative detection is 1.0 ng for both compounds.