Objectives: A sensitive, selective and rapid HPLC method was developed and validated for sodium hyaluronate quantification. Methods: Chromatographic separation was achieved SCHARLAU C₁₈ column (4.6 x 250 mm, 5µm) at 50°C. The mobile phase consisted of 50 mM phosphate buffer (pH = 7) (100 %, v/v) and at a flow rate of 0.5 mL/min. The column eluent was monitored at 205 nm. The retention time (R_t) of sodium hyaluronate was at 3.17 min. Results: The calibration curve was linear over the concentration range of 80 – 320 µg/mL. The intra- and inter-day reproducibility studies demonstrated accuracy and precision according to ICH guidelines. Conclusion: The developed method can be applied to the analysis of sodium hyaluronate in Hycarenol^® eye drops.