The enantiomeric separation of the antidepressant drug fluoxetine and its main metabolite (norfluoxetine) was achieved by using capillary electrochromatography employing packed capillaries with a vancomycin stationary phase. The capillary (21, 23, or 25 cm×75 m ID), with frits at both ends, was connected to a commercially available high sensitivity UV detection cell to improve the detection limit. With the aim of optimizing the separation of the two pairs of enantiomers in the same run, several parameters were studied: these included the mobile phase composition (buffer pH, organic solvent ratio), the capillary temperature, the length of the packed capillary, and the packed stationary phase type. Experiments were performed using a mobile phase containing aqueous ammonium acetate pH 6 (5 mM final concentration) dissolved in 90% acetonitrile/methanol (MeCN/MeOH) with various organic modifier ratios. Increasing MeOH concentration improved the enantioresolution for both analytes. However, the two compounds were not separated under these conditions. The aqueous electrolyte of pH 6 was dissolved at the same concentration as described above in organic solvent (MeCN/MeOH, 70/20 v/v) and used for further studies in which the length of the stationary phase and its composition were changed. Capillaries of 21, 23, and 25 cm length were packed bed length with vancomycin-diol mixed with silica (3:1) or with vancomycin-diol only and tested for the separation of Flx and NFlx enantiomers. Limits of detection (LOD) and limits of quantification (LOQ) as low as 25 and 50 ng/mL, respectively, were observed for each racemic analyte.