Further Studies on Biochemical Active Substances in Urine, Increasing With Fatigue Due to Physical Exercise

Abstract

Abstract-P3and P4 substances were collected from fresh urine of normal male subjects by the techniques of saturation with acetone and precipitation with lead acetate. In addition, FII was fractionated out chromatographically from P3, and was found to have a strong activity in producing cardiac arrest in the frog. It was possible to obtain evidence that the activity of FII is the essence of that of P3 and that FII consists of a single or nearly single substances as chromatographically shown. FII fraction appears to be a peptide of low molecular weight with lysine and tyrosine among the constituents.P4 had no such activity as FII or P3 but suggested the glucosidetype bond. Treatment with acids and heat showed that P4 originates from the same substances as glucuronic acid. Partial hydrolysis of FII resulted in two different ninhydrin-positive components in addition to lysine and tyrosine. Two ninhydrinpositive substances were found among the partial hydrolytes of P4. It is suggested that there is a resemblance in basic structure between FII and P4.There is a variety of in vivo metabolites excreted in the urine. During the course of a disease or following drug administration substances not found physiologically are seen. There is also a ratio variation between the different constituents.It is a well established fact that there are some biochemical active substances present in ordinary human urine and that these can be precipitated by addition of acetone. Acetone insoluble substances (AP) include 2 substances, one precipitated by addition of saturated lead acetate solution (P3) and the other by lead subacetate solution (P4). These biochemical active substances in urine and serum increase during physical exercise. These substances arrest the cardiac beat in the frog. In a previous study (1) an attempt was made to elucidate the essence of these substances with the result that these were found to be proteins of low molecular weight. Using the same procedure, an attempt was made for a refinement utilizing a different technique of analysis. The results obtained are discussed herein. METHODS AND RESULTS1-1. Five liters of fresh urine from males was mixed with 10 1 of acetone. The precipitate was obtained by centrifugation (3,000 r.p.m.), then washed once with acetone. The acetone precipitate (AP) was treate d as shown in Table 1 in order to obtain fraction P3