Annona vepretorum is endemic from the Brazilian Caatinga biome and is used in human nutrition. The present study aimed to investigate the toxic effects of the ethanolic extract from the leaves of this species. The leaves of A. vepretorum were collected, dried, pulverized, and macerated with ethanol to yield the crude ethanol extract of A. vepretorum. HPLC-diode array detection was used to determine the fingerprint chromatogram of the extract. In toxicity studies, the acute toxicity experimental group was administered a single dose of the ethanol extract of A. vepretorum (1 g/kg), while in the subacute toxicity experimental group, the ethanol extract of A. vepretorum was administered orally, daily for 30 days, at doses of 100 and 400 mg/kg. Death and signs of toxicity were observed and at the end, the animals were anesthetized, and blood and organs were then collected. The presence of the flavonoid rutin in the extract was confirmed using HPLC-diode array detection. In the evaluation of acute and subacute toxicity, there were no behavioral and physiological changes or signs of toxicity, and no occurrences of mice deaths were registered. The organs had normal color and preserved architecture, and no statistical variations in weight were observed. The results of the hematological and biochemical parameters after the administration of the ethanol extract of A. vepretorum showed no significant change, except in the count of the number of leukocytes and triglycerides. The histopathologic analysis of the liver, kidneys, and stomach indicated architecture with normal aspects. Thus, the toxicity study indicates low toxicity of the ethanol extract of A. vepretorum. Such information will be helpful in future clinical studies.