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Enantiomeric analysis of chiral drugs is very significant, as their enantiomers display different pharmacological or toxicological behavior towards living systems. Among these drugs, β-blockers are available as racemates, where their enantiomers display different pharmacological effects. Herein, we report enantioselective separation of two β-blockers, namely, atenolol and sotalol, using a derivatization approach. The analytes were derivatized with "(S)-1-[1H-benzo(d)(1,2,3)triazol-1-yl]-2-[6-methoxynaphthalen-2-yl-propan-1-one]" {(S)-BTMNP} in a straightforward derivatization step. The resulting diastereomers were separated on a reverse-phase HPLC C18 column with a mobile phase composed of acetonitrile and TEAP buffer (75:25, v/v, pH = 3.5) and detection at 230 nm. This method achieved successful enantiomer separation for both drugs within 20 min, yielding resolution values greater than 3.8. The detection limits were determined to be 6.4 and 4.6 ng mL -1 for atenolol and sotalol, respectively, which indicated sensitivity and effectiveness of the method for the analysis of two β-blockers from their dosage formulations., (© 2025 Wiley Periodicals LLC.)