Ocimum gratissimum is used in popular medicine to treat painful diseases. The antinociceptive properties of O. gratissimum essential oil (OgEO) and two of its active principles (eugenol and myrcene) were tested in classic models of pain (hot plate test and formalin test). Adult male C57BL/6 J mice acutely received corn oil (control group, p.o.), morphine (positive control group, 5 mg/kg, i.p.), OgEO (10, 20, or 40 mg/kg, p.o.), eugenol or myrcene (both at 1, 5, or 10 mg/kg, p.o.). The highest doses of all tested drugs significantly increased the latency to lick the paw(s) in the hot plate test compared with the control group. OgEO at a dose of 40 mg/kg and eugenol and myrcene at a dose of 10 mg/kg were effective in minimizing animal pain in the first and second phases of the formalin test. The antinociceptive effect shown by all drugs tested in hot plate test was reverted by naloxone administration (1 mg/kg), indicating opiod system participation. These results demonstrate the beneficial effects of OgEO and its active principles against neurogenic and inflammatory pain. Our findings demonstrate that OgEO and its isolated active principles exhibited antinociceptive activity in murine pain models. Copyright © 2012 John Wiley & Sons, Ltd.