The present study focuses on the optimization and validation of a simple, accurate, and precise Reverse Phase High-Performance Liquid Chromatographic (RP-HPLC) method for the quantification of betaxolol hydrochloride in bulk drug and nanoformulations. The chromatographic separation was achieved using a C18 column with a suitable mobile phase, maintaining an optimal flow rate and detection wavelength. The method was validated as per ICH guidelines for parameters including linearity, accuracy, precision, specificity, robustness, limit of detection (LOD), and limit of quantitation (LOQ). The method exhibited excellent linearity within the selected concentration range, with a correlation coefficient (R²) close to 1. Results indicated high recovery and low %RSD values, confirming the method's accuracy and precision. The developed RP-HPLC method is suitable for routine quality control and stability analysis of Betaxolol hydrochloride in both bulk drug and nanoformulation dosage form