Nebivolol is an antihypertensive drug, adrenergic β1 block which is commercialized in the racemic form as RSSS and SRRR-nebivolol. These both enantiomers have distinct pharmacological properties, which suggests a different pharmacokinetic for each one. Therefore, a method to the enantiosselective analysis of nebivolol by capillary electrophoresis was developed and optimized using a derivatized cyclodextrin (Carboxymethyl-β-cyclodextrin – CM-β-CD) as chiral selector. Many parameters were evaluated, such as cyclodextrin and buffer concentration, pH, temperature and voltage applied, in order to optimize the resolution and analysis time. The analyses were performed in a Capillary Electrophoresis system (Beckman Counter - P/ACE™ MDQ) equipped with a DAD operating on the wavelength of 280 nm. After the evaluation of the analytical conditions, it was possible to select the most adequate ones to the resolution of nebivolol enantiomers, which were an electrolyte solution composed by Sodium Acetate buffer 50 mmol L-1, pH 4.0, CM-β-CD 12.5 mmol L-1 and 1% Acetonitrile at 25° C, applied voltage of 25 kV and a 50 µm i.d and 38.0 cm to the detector length capillary. The resolution under these conditions was 1.27 and the migration times for RSSS e SRRR- nebivolol, respectively, were 7.65 and 7.73 min.