To elucidate precise nature of urinary glycosaminoglycans (GAG), GAG in the acidic subfractions obtained previously were quantitated by digestion with mucopolysaccharidases, nitrous acid treatment and by quantitative cellulose acetate membrane (Separax) electrophoresis. Hyaluronic acid, chondroitin sulfates and condroitin, dermatan sulfate and dermatan, heparan sulfate, keratan sulfate and acidic glycopeptide were found in wide distribution in these subfractions. The results suggested the presence of various degree of the depolymerization products of each GAG and of wide range of the desulfation products of each sulfated GAG. Distribution map of each GAG in the subfractions showed more detailed polydispersity of GAG in normal human urine than that reported previously.