Abstract: Garcinia mangostana-Linn (GM) or commonly known as Manggis by Indonesian, is a tropical tree native to Southeast Asia. It produces a fruit, whose pericarp contains tricyclic isoprenylated polyphenol or xanthones. The toxicity’s evaluation of the ethyl acetate fraction from GM pericarp extract was needed before therapeutic use. The purpose of this article is to examine the oral acute toxicity test. The acute toxicity test was conducted in female sprague-dawley rats as per standard protocol. One group of rats administered single dose of 8 mg kg–1 b.wt. and the second group with a single dose of 18 mg kg–1 b.wt., of ethyl acetate fraction of GM extract, administrated orally and one control group. Body weight, behavioral changes and mortality were observed for 14 days. Dietary and water intake were also noted. At the end of the study, rats were sacrificed by decapitation. The result of this study showed that the Lethal Dose (LD50) was found to be >15.480 mg kg–1 b.wt. There was a significant weight increase (p<0.05). Neither mortality nor behavioral changes were noted during 14 day study periods. This study demonstrated that there is a wide margin of safety for ethyl acetate of GM extract and there is no significant toxicity effect on the rats. In conclusion, ethyl acetate fraction GM may be considered for the therapeutic use in pharmaceutical formulation.