Background: Traditional medicine and supplements including Phaleria macrocarpa or Mahkota Dewa are extensively utilised. Rape fruit of phaleria macrocarpa contains flavonoid as a secondary metabolite that has a beneficial effect on endometriosis treatment and other illnesses. The purpose of this study was to look into the subacute toxicity of flavonoid from phaleria macrocarpa fruit extract on the brain and heart of BALB/c female mice. Method: The BALB/c female mice aged 6-8 weeks with a body weight of 20-30gr were divided into four groups and treated for 28 days. The control group received simply water, whereas the treatment group received flavonoid at doses of 500mg/kg/day, 1000mg/kg/day, and 2000mg/kg/day. At the end of the experiment, the mice were sacrificed and important organs were extracted and assessed. The organ weight was assessed using macroscopy, and the brain and heart were assessed using microscopy (histopathological abnormalities). Result: The BALB/c female mice treated with oral administration flavonoid from phaleria macrocarpa fruit extract showed no clinical abnormalities and no change in organ weight compared to the control groups (p>0,5) in subacute toxicity experiments. There were no abnormalities in the organs of the treated mice at a dose of 500mg/kg/day, according to histopatological testing. Conclusion: As a result, flavonoid had no harmful impact in brain and heart mice after subacute therapy up to a level of 500 mg/kg/ body weight. High doses of 1000 and 2000 mg/kg/day are not recommended for long-term use.