Constitutional difference of the acidic glycosaminoglycans (AGAG) in the three layers, the intima, media and adventitia, of normal human aorta was investigated by electrophoretic characterization and enzymatic analyses with other chemical determinations. The anticoagulant activity of the AGAG in the three layers of human aorta was compared by thrombelastography. 2) The analytical data indicated that the AGAG in the three layers out consisted of chondroitin 4-sulfate, chondroitin 6-sulfate, dermatan sulfate, heparan sulfates and hyaluronic acid. It was found that the proportion of dermatan sulfate and heparan sulfates was increased in order from the intima to the adventitia. 3) The predominant anticoagulant actihan in the inner layers referred to the concentration of glucuronic acid on thrombelastogram. The phenomenon was evidenced to be related with the constitutional difference of the AGAG in the three layers.