Determination of fluoxetine and norfluoxetine by high-performance liquid chromatography

A high-performance liquid chromatographic assay was developed for a recently introduced atypical antidepressant, fluoxetine and its demethylated metabolite, norfluoxetine. Prior to analysis, aliquots of alkalinized plasma were extracted with n-hexane and isoamyl alcohol, followed by back-extraction with diluted phosphoric acid. These extracts were injected into a 10 microns, reversed-phase C18 column with phosphate and acetonitrile as the mobile phase and detection at 214 nm. Peak height ratios were linearly correlated up to 800 micrograms/l. Acceptable coefficients of variation were demonstrated for both within-run and day-to-day studies. Selected drugs were checked for interference. The assay was used to monitor nine patients receiving 20 to 80 mg of fluoxetine per day. Plasma concentrations of fluoxetine and norfluoxetine ranged from 37 to 301 micrograms/l and 29 to 326 micrograms/l respectively.