Establishment of an HPLC Method for Simultaneous Determination of Chondroitin Sulfate and Hyaluronic Acid in Cartilage Hydrolysate
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Abstract:
To accurately and rapidly determine the contents of chondroitin sulfate and hyaluronic acid in cartilage hydrolysate, galactosamine and glucosamine were used as the target substances. A determination method based on high-performance liquid chromatography was developed by optimizing the chromatographic conditions and methodological tests. The ZORBAX C18 column was used for separation, with acetonitrile and ammonium acetate solution as the mobile phase for gradient elution. The flow rate was 1.00 mL/min, and the measurement wavelength was 250 nm. Methodological tests showed that the mass concentration of chondroitin sulfate and hyaluronic acid followed a good linear relationship with the peak area in the range of 100~500 mg/L. The limit of detection of chondroitin sulfate was 0.948 mg/L, its repeatability deviation was 0.089%, average recovery rate was 99.71%, and relative deviation was 0.35%. Meanwhile, the limit of detection of hyaluronic acid was 0.027 mg/L, its repeatability deviation was 0.045%, average recovery rate was 97.74%, and relative deviation was 0.92%. Compared with the conventional method, the proposed method offers the advantages of being less affected by interference from impurities and having high accuracy, excellent repeatability, high recovery rate, and high precision. Using this method, the contents of chondroitin sulfate and hyaluronic acid in cartilage hydrolysate can be detected rapidly and accurately.
