Objectives: This study was performed to observe the toxicological changes caused by a single exposure to benzo[a]pyrene. Methods: Based on the results of a preliminary study, 300 mg/kg was set as the middle dose. A highest dose of 2,000 mg/kg and a lowest dose of 50 mg/kg were selected based on GHS guidelines. Benzo[a]pyrene was orally administered once to female and male SD rats at dose levels of 50, 300, and 2,000 mg/kg (body weight). All animals were monitored daily for clinical signs and mortality over 14 days. Hematological and biochemical values were examined as well. Results: There were neither dead animals nor significant changes in body weights during the experimental period. In addition, no differences were found between the control and treated groups in clinical sign, hematology, serum biochemical, and histopathological analysis. Conclusion: Compared with the control group, we could not detect any toxic alteration in all treated groups. These studies indicate that the acute toxicity of benzo[a]pyrene is relatively low.