The aim of the study was to evaluate the experimental acute and sub-chronic toxicities of Operculina macrocarpa with plant extract and powder, respectively, in male and female mice. Phytochemical prospection was performed with extract and administration in single doses by intraperitoneal route to six groups and control at 1230, 970, 700, 350, 120 and 30 mg/kg doses and distilled water (0.1 ml/10 g), respectively. Sub-chronic doses of 1230, 700, 30 mg/kg/day and satellite (1230 mg/kg/day) were administered orally in feed. Major endpoints included alterations in the central and autonomic nervous system, water and food intake, body weight, hematological and biochemical parameters. Phytochemical screening identified compounds: Alkaloids, flavonoids, xanthones, leucoanthocyanidins and tannins condensate. In the acute study, mortality was observed with toxicity signs to the central nervous system (CNS) at LD 50 of 270 mg/kg. There were no significant changes in water and food intake, body weight, haematological and biochemical parameters, and histopathological examination in the subchronic study (p value). Results indicate that the oral administration of O. macrocarpa powder in feed is less toxic and relatively safer.