It is well known that serum albumin, γ-G-globulin and glycoprotein increase in urine after physical load. There are two ways, the Donaggio reaction and ASPRO method (a new acid soluble glycoprotein assay using coomassie brilliant blue G-250), to measure glycoprotein and other acid soluble protein for estimating the physical load. This investigation was performed to identify the composition of protein detected by the Donaggio reaction (DRPSu) and of perchloric acid soluble protein detected by the ASPRO method in urine (ASPROu). In DRPSu, albumin, α1-acidglycoprotein, α2-HS-glycoprotein, Zn-α2-glycoprotein, α1-antitrypsin, α1-antichymotrypsin and transferrin were recognized by double diffusion, single radial immunodiffusion and immunoelectrophoresis. In ASPROu, the same proteins as above were recognized according to the same immunological methods. However, a different ratio of protein components was recognized between DRPSu and ASPROu by SDS-PAGE. The perchloric acid soluble protein in urine before and after exercise from high school boys was measured by the ASPRO method, and a significant difference was recognized between before and after exercise.