Phytolacca americana is a species of the Phytolaccaceae family native to North America. P. americana is used as a folk medicine to treat inflammation, fungal infections, rashes and breast problems; moreover, recent publications support possible anticancer properties. The purpose of this study was to characterize and compare the phytochemical constituents of the leaves of P. americana, and determine their antifungal potential. For this, three temperatures were evaluated to dry the leaves: ambient temperature (12 ± 2 °C), 22 °C and 40 °C, then extracts of the leaves were obtained by ethanol extraction. FTIR and UV/VIS spectroscopy analysis of the extracts shows that the drying temperatures do not affect the stability of the biosynthetic groups. Main secondary metabolites such as flavonoids, saponins, coumarins and tannins were determined by phytochemical screening. Furthermore, extracts were analyzed by means of gas chromatography coupled to mass spectrometry (GC-MS), essential acids such as octadecanoic and some alcohols were identified. Finally, the antifungal potential of ethanolic extracts with concentrations of 60 to 300 mg/mL was evaluated in four types of strains: F. solani, A. brasiliensis, S. kudriavzevii and C. albicans. Results shows inhibition diameters of 18 and 19 mm for F. solani and C. albicans strains respectively.