Red fruit (Pandanus conoideus Lamk) empirically has been used Papuapeople , Indonesia as a natural medicine to treat a variety of diseases including cancer, HIV / AIDS, herpes and diabetes. The information about the toxicity of this plant is very important considering this plant is potential as medicine. The aim of this study is to determine acute toxicity of red fruit oil and its effect on hepatic enzyme level (AST and ALT). The oil was extracted by heat extraction method. Acute toxicity testing conducted based on OECD 423 guideline. In acute toxicity study, the oral dose of red fruit oil was administrated to 3 group (300, 2000 and 5000 mg/kg BW) in single dose. The general behaviour, adverse effect and clinical symptom was observed every hour in first 4 hours, 24 hours,48 hours, and continue to observe for 14 days after administration of red fruit oil. No animals showed toxic symptoms in 300 and 2000 mg/kg dosing group. One animal in 5000 mg/kg BW dosing group had diarrhea one hour after administration. No animal dead in this experiment after 14 days observation. AST and ALT mean value for rats on 300 mg/kg BW, 2000 mg/kg BW, and 5000 mg/kg BW dosing groups are 22.70±1:05 IU/L , 24.15±8.89 IU/L and 24.54 ± 6.26 IU/L and 18.04 ± 0.77 IU / L,19.69 ± 3.08 IU/L, 16.78±1.60 IU / L, respectively. No statistically significant difference of the value of AST and ALT levels in each treatment group (p>0.05). Based on the 2001 OECD acute toxicity, red fruit can be categorized as Category 5 GHS (Globally Harmonized System for Chemical Classification Subtances and Mixtures) as practically non-toxic materials.