Due to its therapeutic action, the phytochemical study of the essential oil of Aniba canelilla H.B.K Mez. (Lauraceae) and its toxicological preclinical trial were evaluated. The essential oil extracted from the bark (OEAC) yielded 1.0% and the major constituents were 1-nitro-2-phenylethane (52.9%) and methyl eugenol (38.7%), both identified by GC/ MS and GC-FID. The observed LD 50 (oral) was 720 ± 66.4 mg/kg. The doses used in the subacute toxicity study (1/10 and 1/20 of the LD 50 ) were administered in rats for 30 days, and the biochemical (glucose, urea, creatinine, triglycerides, cholesterol, glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase and alkaline phosphatase) and hematological (erythrocytes, leukocytes and platelets counts, blood hemoglobin concentration, hematocrit and mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration) parameters were observed. The results suggested that the A. canellila oil presented low toxicity in rats, according to the used dosages.