The anxiolytic effect of Eurycoma longifolia Jack in mice was examined. Fractions of E. longifolia Jack extract produced a significant increase in the number of squares crossed (controls=118.2±10.2 squares), but significantly decreased both the immobility (controls=39.4±4.0 sec) and fecal pellets (controls=12.3±2.1 fecal pellets) when compared with control mice in the open-field test; they significantly increased the number of entries (controls=6.7±0.5 entries) and time spent (controls=42.9±0.1 sec) in the open arms, but decreased both the number of entries (controls=13.2±0.7 entries) and time spent (controls=193.4±0.7 sec) when compared with the control mice in the closed arms of the elevated plusmaze test. Furthermore, fractions of E. longifolia Jack extract decreased the fighting episodes significantly (controls=18.0±0.4 fighting episodes) when compared with control mice. In addition, these results were found to be consistent with anxiolytic effect produced by diazepam. Hence, this study supports the medicinal use of this plant for anxiety therapy.