Urospatha sagittifolia is a medicinal herb commonly used by the indigenous communities of the Amazon. It is known to be effective in treating venomous snake bites, as well as hemorrhagic and inflammatory conditions. This study aimed to know the chemical composition of U. sagittifolia tubers at different growth stages, using LC-MS and GC-MS metabolomics and multivariate analysis. Growth stages of U. sagittifolia were categorized as: seedlings (0.20 m-height), juveniles (0.20-1.15 m-height), and adults (> 1,15 m-height with reproductive organs). We identified 19 metabolites with authentic standard (level 1 identification). Also, we tentatively identified 24 metabolites using public spectra libraries in GNPS (level 2 identification). Additionally, GC-MS allowed the tentative identification of 7 metabolites. Based on peak height, fatty acids such as methyl palmitate and methyl stearate were found to be the most abundant in lower growth stage, while organic acids such as allantoic acid and palmitic acid prevailed as the plant matured. We also found that juvenile´s tubers contain mayor metabolites, and its relative concentration is more than adult or seedling tubers. This is the first report about differences in secondary metabolite composition at different growth stages of U. sagittifolia which can be beneficial when used in a particular treatment.