Objective: In the past decade, dissolution testing has emerged as a valauble tool for the characterization of drug product performance in the field of pharmaceuticals. During the development of new formulations, dissolutions tests assist in the evaulation of any changes in the formulation arising during manufacturing process, thereby assuring product quality and performance post manufacturing.Methods: In the present study, a simple high performance liquid chromatography (HPLC) method was developed and validated to quantitate the release of L‐Dopa from poly (D, L‐lactic‐co‐glycolic acid) (PLGA) nanoparticles. The chromatographic separation was performed with a reversed‐phase C18 column, using acetonitrile‐water containing 0.05% trifluoroacetic acid (5:95, v/v) as a mobile phase at 280 nm. The developed method was validated for its specificty, linearity, accuracy, and precision according to the ICH guidelines.Results: The developed method was shown to be linear (r2 ≥0.995) in the concentration range of 125-40 µg/mL. The mean % recoveries were found to be 102.59-98.70%, indicating an agreement between the true value and the detected value. Solution stability was guaranteed by the addition of an antioxidant. The analytical method was shown to be suitable for the evaluation of release of L‐Dopa from PLGA nanoparticles. In vitro release of L‐Dopa was studied using sample and separate (SS) and dialysis membrane (DM) methods. To compare SS and DM methods, difference (ƒ1) and similarity (ƒ2) factors were calculated. No significant differences were recorded in the release kinetics of L‐Dopa from nanoparticles using both methods (ƒ1<15 and ƒ2>50).Conclusion: Dissolution test methods were compared and procedure for an analytical method based on HPLC was optimizated and validated for the dissolution of L‐Dopa loaded nanoparticles.