The principal goal of this work was to investigate the liquid chromatographic retention behaviour of fluoxetine and norfluoxetine using HPLC with respect to mobile phase composition, pH, flow rate and the amount of the native β-cyclodextrin (β-CD) or the β-hydroxypropyl-cyclodextrin (HP-β-CD), added to the mobile phase. Further, it was of interest to evaluate the effectiveness of β-CD and HP-β-CD to enhance fluorescence detection of these compounds.