Eurycomanone has been identified as the major bioactive compound contributing to Eurycoma longifolia (EL) aphrodisiac activity, however, its mechanism of action remains obscured. Presently, eurycomanone was isolated from EL root extract and its molecular structure was identified. The human neuroblastoma SH-SY5Y cell line was differentiated into human dopaminergic neuron-like cells. Exogenous dopamine levels from the differentiated SH-SY5Y cells were quantified following the treatment of 5, 10, 15 μM of eurycomanone and 10 μM clorgyline as positive control. Dopamine secretion was significantly increased in a dose-dependent manner, compared to the vehicle control (p