Objective: The main objective of present study was to develop and validate a reverse phase chiral high performance liquid chromatographic method for R & S enantiomeric separation of Lercanidipine hydrochloride in API as well as tablet dosage form. Methods: The R-Isomer of Lercanidipine Hydrochloride was resolved on a Chiral OJ-H (150 x 4.0mm, 5um ) column using mobile phase system containing 10mM Ammonium acetate and Acetonitrile in the ratio of (35:65 v/v). The mobile phase was set flow rate of 1.0ml and the volume injected was 10ul for every injection. The detection wavelength was set at 240nm. Results: The proposed method was productively applied for the quantitative determination of (R)-Isomer in Lercanidipine hydrochloride in API as well as tablet dosage form. The linear regression analysis data shows good linear relationship over a concentration range 0.5ug/ml to 4ug/ml for (R)-Isomer. The mean value of the correlation coefficient was 0.998 for (R)-Isomer. The method was validated as per the ICH guidelines. The detection limit (LOD) was about 0.05ug/ml and quantisation limit (LOQ) was about 1.0ug/ml for (R)-Isomer. The relative standard deviation was found to be 2.0% for (R)-Isomer of Lercanidipine Hydrochloride. Conclusion: The method is specific, rapid, precise and accurate for the separation and determination of (R)-isomer in (S) - Lercanidipine hydrochloride in API & tablet dosage form. The developed and validated HPLC method and the statistical analysis showed that the method is repeatable and selective for when compared to earlier published research articles for the estimation of (R)-Isomer of the Lercanidipine Hydrochloride drugs substances as well as in tablet dosage form.