Nadolol (SQ 11725) is a β-adrenergic blocker which has two chiral centers, this allows for the presence of two racemates known as racemate A (SQ 12181) and racemate B (12182) and a total of four enantiomers. A simple isocratic HPLC method is developed for the separation of nadolol racemate A (SQ 12181) to its corresponding enantiomers RSR-nadolol (SQ 12148) and SRS-nadolol (SQ 12150) and racemate B (SQ 12182) to its corresponding enantiomers RRS-nadolol (SQ 12149) and SSR-nadolol (SQ 12151). Hexane with different percentage of ethanol containing 0.4% diethylamine is used as a mobile phase.