An UPLC-HR-MS metabolomics approach was used to study the effects of a 49-days oral supplementation with Polygonum cuspidatum extract in healthy rats. Multivariate analysis allowed to observe significant differences in the excretion of several markers between treated animals and control group. Among the others, the amounts of N-methyl-2-pyridone-5-carboxamide (2PY) and phenylacetylglycine (PAG) were reduced in the treated group compared to control. These compounds have been previously considered as markers of aging. Furthermore, the excretion of 3-hydroxysebacic acid and 4,6-dihydroxyquinoline was also changed following supplementation, although not significantly. Despite the relatively short time of treatment (7 weeks), the significant changes in the urinary levels of aging markers observed at day 49 suggests a potential role of this type of studies as a new approach in the evaluation of the anti-aging effects of plant extracts.