To obtain a clue for the metabolic disorder of glycosaminoglycans (GAG) in connective tissue diseases (CTD), a device of screening test for excretion pattern of urinary GAG was achieved. The crude GAG were separated from voluntary 100-20 ml urine by precipitation with cetylpyridinium chloride, followed by digestion with pronase P. The crude GAG were then separated by one-dimensional Separax (cellulose acetate membrane) electrophoresis in 0.2 M aqueous calcium acetate. GAG, in terms of hexosamine, in each GAG-band were quantitated after extraction from the strip with water. All the urines examined gave three regular bands, bands 1, 2 and 3, but urines from most of the pupils at puberty and from several adults gave irregular bands, besides the regular ones. As for the regular bands, a significant difference was observed in percent distribution of bands 1, 2 and 3 between the samples from male and female of the normal youth aged 20-29, and excretion pattern of three regular bands reached to an adult type much younger in female than in male. GAG in the regular bands were identified as follows: band 1, partially desulfated chondroitin sulfates A and C; band 2, dermatan sulfate plus low-sulfated chondroitin sulfates (LSC) in most cases; band 3, heparan sulfate plus LSC in most cases. Usefulness in clinical field of the screening test is indicated.