A simple, rapid and robust LC method was developed and validated for the enantiomeric separation of dexrabeprazole in bulk drug and formulation. The enantiomers of dexrabeprazole were resolved on a Chiralpak AD-RH (amylose based stationary phase) column using a mobile phase consisting of water: acetonitrile (50:50, v/v) at a flow rate of 0.5 ml min¹. The resolution between the enantiomers was found to be more than 1.5 in optimized method. The developed method was extensively validated and proved to be robust. The calibration curve for (S)-enantiomer showed excellent linearity over the concentration range of 0.05 µg ml¹ (LOQ) to 1 µg ml¹. The limit of detection and limit of quantification for (S)-enantiomer were 0.015 µg ml¹ and 0.05 µg ml¹, respectively. The percentage recovery of the (S)-enantiomer ranged between 97 to 101 % in bulk drug samples of dexrabeprazole. The proposed method was found to be suitable and accurate for quantitative determination of (S)-enantiomer in bulk drug substance.