The present study reports the molecular dereplication and the evaluation of anti-Trypanosoma cruzi activity of volatile oils from inflorescences, leaves, branches, and roots of Saururus cernuus L. (Saururaceae). Chemically, the oils showed the predominance of sesquiterpenes in inflorescences (64.04%) and branches (63.82%), with β-sesquiphellandrene (25.50%) and (E)-caryophyllene (22.40%) corresponding to the main constituents of each oil. Furthermore, it was possible to detect safrole as the most predominant compound in the leaves oil (49.09%). On the other hand, the oil from roots was mainly composed by monoterpenes (84.60%), with limonene in higher concentration (38.41%), followed by α-pinene (20.19%), and camphene (14.71%). The oils from inflorescences and branches displayed higher antitrypanosomal potency with EC50 values of 7.1 and 8.8 μg mL-1, respectively, followed by the oils from the roots and inflorescences, with EC50 values of 17.3 and 30.4 μg mL-1, respectively. Additionally, branches and inflorescences oils displayed no toxicity in mammalian NCTC cells (CC50 > 200 μg mL-1). Using two PLS-DA methods, it was possible to suggest that the anti-T. cruzi activity of the tested oils could be associated with the presence of β-sesquiphellandrene, safrole, β-elemene, and α-zingiberene whereas threo-austrobailignan-5, β-sesquiphellandrene, α-humulene, germacrene D, and bicyclogermacrene play a role in the cytotoxicity against NCTC cells.