A liquid chromatography(HPLC)method with UV detection was developed for determination of sodium hyaluronate in pharmaceutical formulation.Sodium hyaluronate is a polymer of disaccharides,composed of D-glucuronic acid and D-N-acetylglucosamine,linked via alternatingβ-1,4 andβ-1,3glycosidic bonds.Being a polymer compound it lacks a UV absorbing chromophore.In the absence of a UV absorbing chromophore and highly polar nature of compound,the analysis becomes a major challenge.To overcome these problems a novel method for the determination of sodium hyaluronate was developed and validated based on size exclusion liquid chromatography(SEC)with UV detection.An isocratic mobile phase consisting of buffer 0.05 M potassium dihydrogen phosphate,pH adjusted to 7.0using potassium hydroxide(10%)was used.Chromatography was carried out at 25 1C on a BioSep SEC S2000,300 mm 7.8 mm column.The detection was carried out using variable wavelength UV–vis detector set at 205 nm.The compounds were eluted isocratically at a steady fow rate of 1.0 mL/min.Sodium hyaluronate retention time was about 4.9 min with an asymmetry factor of 1.93.A calibration curve was obtained from 1 to 38 g/mL(r40.9998).Within-day%RSD was 1.0 and between-day%RSD was 1.10.Specifcity/selectivity experiments revealed the absence of interference from excipients,recovery from spiked samples for sodium hyaluronate was 99–102.The developed method was applied to the determination of sodium hyaluronate in pharmaceutical drug substance and product. A liquid chromatography (HPLC) method with UV detection was developed for sodium hyaluronate in pharmaceutical formulation. Sodium hyaluronate is a polymer of disaccharides, composed of D-glucuronic acid and DN-acetylglucosamine, linked via alternating β-1,4 and β-1 , 3 glycosidic bonds. Being a polymer compound it lacks a UV absorbing chromophore. In the absence of a UV absorbing chromophore and highly polar nature of compound, the analysis becomes a major challenge. To overcome these problems a novel method for the determination of sodium hyaluronate was developed and validated based on size exclusion liquid chromatography (SEC) with UV detection. An isocratic mobile phase consisting of buffer 0.05 M potassium dihydrogen phosphate, pH adjusted to 7.0 using potassium hydroxide (10%) was used. Chromatography was carried out at 25 1C on a BioSep SEC S2000, 300 mm 7.8 mm column. The detection was carried out using variable wavelength UV-vis detector set at 205 nm. The compounds were eluted isocratically at a steady fow rate of 1.0 mL / min. Sodium hyaluronate retention time was about 4.9 min with an asymmetry factor of 1.93. A calibration curve was obtained from 1 to 38 g / mL (r 40.9998) .Within-day% RSD was 1.0 and between-day% RSD was 1.10. Specificity / selectivity experiments revealed the absence of interference from excipients, recovery from spiked samples for sodium hyaluronate was 99-102. The developed method was applied to the determination of sodium hyaluronate in pharmaceutical drug substance and product.