Quantification of hyaluronan and chondroitin/dermatan sulfates in the tissue sections on glass slides

The method for the determination of hyaluronan and chondroitin/dermatan sul fates in the tissue sections on a glass slide, which were prepared by histo logical technique, was established by applying to porcine skin. The degrada tion of these glycosaminoglycans to the unsaturated disaccharides in porcin e skin sections on a glass slide was achieved by chondroitinase ABC and ACI I in the presence of highly purified bacterial collagenase. Subsequently, t he resulting unsaturated disaccharides were determined by HPLC with fluorom etric postcolumn derivatization using a-cyanoacetamide as a reagent. So far , the determination of the glycosaminoglycans in the tissues has taken up m ore than 5 days, whereas the determination of the glycosaminoglycans in the frozen sections by the present method was completed within a day. In addit ion, applications of the present method to the serial polyester wax section s processed with a small surgical knife made it possible to determine the g lycosaminoglycans in a local part in the tissue section. The present method should open a way for the clinical analysis of glycosaminoglycans in the p athological tissue samples. (C) 1999 Academic Press.