[Analysis of hyaluronic acid in punctates in biochemical functional tests of joints]

The punctates of the biochemical joint function test contain hydroxyethyl starch which disturbs the assay of uronic acid and glucosamine for the estimation of hyaluronate. Therefore hyaluronate (0.03-0.75 g/l) was estimated by precipitation with Alcian blue on cellulose acetate membranes and measuring of the absorbance at 678 nm after dissolution in aqueous SDS (75 g/l). The gel-filtration HPLC separates the high-molecular-mass hyaluronate from low-molecular-mass impurities of biological samples. By comparing with a standard curve (0.5-2.5 g/l hyaluronate) the concentration can be determined. The results obtained by the two methods were significantly correlated (p less than 0.05). The Alcian blue technique is useful for rapid screening, the HPLC technique to confirm these values.