Analysis of the Enantiomers of Fluoxetine and Norfluoxetine in Plasma and Tissue Using Chiral Derivatization and Normal-Phase Liquid Chromatography

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Abstract
A stereospecific HPLC assay has been developed for the determination of the enantiomers of fluoxetine and an active metabolite, norfluoxetine, in plasma and tissue. Following derivatization with R-1-(1-napthyl)ethyl isocyanate, the diasteriomeric derivatives are resolved on a 5 μ.m silica column using a iso-octane/tetrahydrofuran mobile phase with fluorescence detection. The assay has a sensitivity of 5 ng/ml in plasma and 25 ng/gm in tissue. The calibration curves are linear over the range of 5–1000 ng/ml. The assay appears to be readily applicable to the study of enantioselective fluoxetine pharmacokinetics in animals and humans.