The present study was designed to explore the total phenols and flavonoids content in leaf fractions of Plantago australis L., and their radical scavenging activity by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The free radical scavenging abilities of fractions were determined as: ethyl acetate fraction > crude extract > butanolic fraction > chloroformic fraction. Besides having the highest radical scavenging activity, the ethyl acetate fraction also obtained the highest total phenols and flavonoids contents. Chemical profile of this fraction was obtained by high performance liquid chromatography (HPLC) with photodiode array detector. The comparison with the literature for the ultraviolet espectra for the chromatographic peaks allowed finding compounds similar to neolignins, cinnamic acids derivates, flavonoid (luteolin 7-glycoside) and anthocyanic pigments.