It is an analytical requirement to identify and determine high molecular weight APIs in pharmaceutical forms. For this reason, it is aimed to develop and validate an analytical method for the determination of hyaluronic acid in pharmaceutical form in this study. 0.1 M Na2SO4 was prepared as mobile phase. The separation of compound was performed with a OHpak Shodex SB-806M HQ (13 µm, 8,0 x 300mm, Japan) analytical column. Guard column which is OHpak SB-G, (8 µm, 100 Å, 6.0 x 50 mm, Japan) was integrated to the system before the analytical column. UV detection at 198 nm was used to monitor the eluent and flow rate of the mobile phase was set to 1 mL/min. The method has been validated in terms of system suitability, linearity, limits of detection (LOD) and quantity (LOQ), precision, accuracy, specificity, selectivity, and stability. The obtained findings showed that the analytical method has linearity higher than 0.99, accuracy, precision, selectivity and stability. The method was found to be precise, accurate and specific during the study.