Bioactive natural compounds derived from plants are the source for the development of new drugs. Numerous in vitro studies have explored the anti-inflammatory effect of eurycomanone and fargesin, derived from Eurycoma longifolia and Flosmagnoliae, respectively. However, before anti-inflammatory investigation is conducted, it is important to obtain the safe doses of these compounds to ensure the validity of the anti-inflammatory results. Therefore, the present study was aimed to investigate the cytotoxicity of eurycomanone and fargesin towards macrophage RAW 264.7. cells to determine the safe doses of these compounds. Different concentrations of eurycomanone and fargesin were subjected to RAW 264.7 cells. The cytotoxicity of the compounds was evaluated by MTT assay and 50% inhibitory concentration (IC50) of these compounds was determined. Morphological changes of RAW 264.7 cells upon exposure to these compounds were also observed. Eurycomanone exhibited its cytotoxic effect by reducing RAW 264.7 cell viability dose-dependently with the IC50 of 94.17 µM. Meanwhile, fargesin had slight cytotoxicity towards RAW 264.7 cells with the IC50 of 173.5µM. Eurycomanone was more cytotoxic towards RAW 264.7 cells compared to fargesin. In conclusion, eurycomanone and fargesin at concentration up to 25 µM, was not toxic to the RAW 264.7 murine macrophages cells and the findings can be applied in the future anti-inflammatory study.