While caloric restriction (CR) is associated with a prolonged lifespan in multiple species by regulating metabolism, a comprehensive profile of metabolism under CR conditions remains largely unclear. Therefore, in this study we aimed to characterize the metabolomic profiling associated with CR using a rat model. Rapid resolution liquid chromatography/electrospray ionization quadrupole-time of flight mass spectrometry (RRLC/ESI-Q-TOFMS) was employed to analyze metabolomic profiling of urine samples from aging rats who underwent caloric restriction (CR; n=7) or were provided a normal diet (N; n=8) for 12 weeks time. Multivariate data analysis was performed on the mass data of metabolomic profiles to uncover the differences between the CR and N groups. CR treatment led to manifest metabolic changes in aging rats, and fifteen urinary metabolites including hypoxanthine, hippurate, dimethylglycine and creatinine were significantly different in the rat groups. Our study demonstrates the high reliability of the HPLC-based metabolomic approach towards the study of anti-aging effects induced by CR, while the urinary metabolites we identified may become potential biomarkers of aging.