Liquid-chromatographic determination of hyaluronic acid in pleural and ascitic fluids

Abstract

A previously described method for determination of hyaluronic acid is modified for the analysis of pleural and ascitic fluids. After 20 μL of the acellular supernate is precipitated with ethanol, the precipitate is digested overnight with chondroitinases and chondroitin sulfatases. The content of hyaluronic acid-derived disaccharide is then analyzed by 'high-performance' liquid chromatography. The high sensitivity (detection limit, <0.1 μg of hyaluronate-derived uronic acid per millimeter) and reproducibility (SD = 4% of the mean) enable accurate determination of hyaluronic acid, even in effusions from patients without signs of mesothelioma. The range of values obtained for pleural and ascitic fluids from patients without signs of this kind of tumor is the same as that found by using other techniques, being slightly higher in those patients showing signs of tissue destruction. The concentrations of hyaluronic acid were considerably increased in most, but not all, of the cases of mesothelioma.