Background: Colorectal cancer (CRC) ranks as the third most prevalent malignancy globally, presenting a formidable early diagnostic challenge. An effective biomarker with high sensitivity and specificity can help diagnose CRC and improve the chances of successful treatment. Methods: 100 healthy controls and 95 CRC patients( 25 Stage 0/I,30 stage II and 40 stage III based on Clinical stages) were recruited. Subsequently, 195 urine samples were subjected to UPLC-MS analysis. Comparative analysis was employed to elucidate noteworthy metabolic variances, and pathway analysis was conducted to unveil perturbed metabolic functions. Ultimately, metabolic panels for CRC diagnosis were constructed. Result: A total of 82 metabolites exhibited statistical significance between CRC patients and healthy controls. Moreover, pathway analysis revealed that they were associated with Steroid hormone biosynthesis, Nitrogen metabolism, and D-Glutamine and D-glutamate metabolism. A composite panel consisting of Retinol, L-β-aspartyl-L-glycine and 21-Deoxycortisol showed AUCs of 0.933/0.93 in the discovery/validation group. The panel also showed commendable efficacy in indifferent CRC stages, with an AUC of 0.918 for stages 0/I, 0.862 for stage II, and 0.845 for stage III. Conclusions: Urine metabolomecould distinguish CRC from healthy control and reflect the changes in different stages of CRC. Potential biomarkers might be developed by targeted metabolomic analysis.