HPLC Determination of EDTA in Hyaluronic Acid

HPLC was applied to the determination of EDTA in hyaluronic acid. ZORBAX Eclipse XDB-C18 column (4.6 mm×150 mm, 5 μm) was used as stationary phase and a mixture of acetonitrile, 100 g·L-1 tetrabutyl ammonium hydroxide of pH 6.5 and water (20 to 20 to 60 by vol.) was used as mobile phase. UV-detection at the wave-length of 260 nm was adopted. Linear relationship between values of peak area and mass concentration of EDTA was kept in the range of 0.037 2-1.86 mg·L-1, with detection limit (3S/N) of 4.745 μg·L-1. The proposed method was used in the analysis of sample hyaluronic acid, values of recovery found were in the range of 87.2%-93.6% with RSD′s (n=5) less than 5%.