Ranunculus macrophyllus Desf. is a multipurpose edible medicinal plant, whose roots are used to treat feminine infertility and to increase weight. The aim of this work was to study for the first time the acute toxicity and anti-inflammatory activity of their n-hexane extract. Gas chromatography was used to study the chemical composition of the extract, while xylene induced ear edema was used to assess its anti-inflammatory activity. In silico methods were used to determine the possible anti-inflammatory mechanism of action and pharmacokinetic behavior of the major compounds. Gas chromatographic analysis led to identification of 19 fatty acids, including linoleic, ¿-linolenic and palmitic acids as major compounds. The n-hexane extract did not induce any visible signs of toxicity or mortality to mice at 2000 mg/kg body weight, and successfully inhibited the xylene induced ear inflammation (67% at 200 mg/kg). This activity may be attributed to linoleic and ¿-linolenic acids that were found to be potent inhibitors of cyclooxygenase 2 and phospholipase A2 in the in silico study. The n-hexane extract exhibited strong anti-inflammatory activity with no acute toxicity, which opens the doors for the use of R. macrophyllus roots and their n-hexane extract in food, nutraceutical and pharmaceutical industries.