Nadolol is a blocking agent with activity in β-adrenergic receptors. The objective of this research was to develop and validate an HPLC and circular dichroism method for the quantification of four nadolol stereoisomers in capsules. The HPLC method was validated using a Chiralcel OD column (250 × 4,6 mm, 10 μm). The mobile phase consisted of hexane-ethanol-diethylamine-acetic acid (86 + 14 + 0.3 + 0.3, v/v/v/v), with a flow rate of 0.7 mL/min and temperature of 25 ± 1°C and UV detection carried out at 220 nm. Solutions were prepared in ethanol containing 200 μg/mL nadolol. The method proved to be precise, selective, accurate, and robust and was successfully applied for the determination of the two homologs of four nadolol stereoisomers in capsules.