The extracellular conditioned medium of MCF-7 breast cancer cells treated by DMSO and cow urine DMSO fraction (CUDF) (50 µg/ml) final concentration was collected as per the previously published procedure . Then, 750 µl of extracellular conditioned medium was mixed with 4X loading buffer (0.5 M Tris, pH 6.8, and Glycerol). Next, the conditioned medium along with the loading buffer was loaded on the vertical tube gel electrophoresis (VTGE) purification system with a matrix of 15% polyacrylamide gel (acrylamide: bisacrylamide, 30:1) . The fractionated extracellular metabolites were collected in the running buffer (96 mM glycine, pH 8.3). The detailed procedure was adopted from previously published in-house VTGE-assisted purification of metabolites . Furthermore, LC-HRMS analyses of VTGE-purified extracellular metabolite elutes were performed by Agilent TOF/Q-TOF Mass Spectrometer station Dual AJS ESI ion source. During LC separation, RPC18 Hypersil GOLD C18 100 x 2.1 mm-3 µm and mobile phase of 100% Water (0.1% FA in water) and 100% Acetonitrile (90% ACN +10% H2O+ 0.1% FA) were used in the proportion of 95% and 5% (40). Mass spectrometry was performed in a positive mode and analyzed as per the procedure adopted from previously reported methodology .