This study is an experimental in vitro research, aimed to evaluate antimicrobial potential and cytotoxicity of plant species Argemone mexicana L. (Papaveraceae) with a treatment of infected wounds perspective. Crude ethanolic extract was obtained after the solution concentration in a rotary evaporator and submitted to a fractionation by classical liquid chromatography column, resulting in four fractions: hexane, chloroform, ethyl acetate and methanol. A phytochemical screening was performed in crude ethanolic extract. Antibacterial activity was determined by the Minimum Inhibitory Concentration (MIC), where fractions were tested against major infectioncausing bacteria in wound and to evaluate the samples cytotoxicity was conducted the MTT (3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium reduction assay. In the studied samples, it was observed the presence of steroids, coumarins, anthrones and alkaloids, which is the majority metabolite. The cytotoxicity of the samples was observed at concentrations of 1000 μg and 100 μg. Among these, chloroform fraction at a concentration of 1000 μg and methanol fraction at a concentration of 100 μg showed no cytotoxicity with cell viability of 87 % and 90, 2 %, respectively. The antibacterial activity was observed in fractions against S. epidermidis and in methanol fraction front S. aureus, which showed a MIC of 166,66 μg/mL. This study may be useful to guide new investigations, depth and systematic, about the chemical compounds of this species and serves as subsidies for further herbal medicine development with perspective in treatment of infected wound.