Plasma glycosaminoglycans (GAGs) were isolated and purified by chromatographic procedures in healthy humans and horses before and after physical exercise. A weak anion exchange resin was used to separate polyanions. Humans exercised on a cycloergometer, while horses were exercised on a treadmill and in show jumping competition. Some GAGs were isolated from untreated plasma and operationally defined as native proteoglycans (PGs), while the total GAG amount was isolated from mild alkali treated plasma. Plasma GAG concentrations in terms of galactosamine and galactose, the respective components of the cartilage polysaccharides chondroitin sulphate and keratan sulphate, significantly increased only when the subject was exercised intensively, in both humans and horses. Only native PGs fraction contributed to GAG levels changes. The increases were transient, since preexercise values were reached in 30 min in horses. These changes with exercise are suggested to be a tool to measure the effect of physical exercise on cartilage PG metabolism.