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Hancornia speciosa Gomes is a fruit tree, commonly known as the mangaba tree, which is widespread throughout Brazil. The leaves of this plant are used in traditional medicine for medicinal purposes. Thus, the objective of this study was to perform a physicochemical characterization, identify the lipophilic antioxidants and fatty acids, and determine the microbiological quality and safety of H. speciosa leaves. In addition, the antioxidant, antimutagenic, and inhibitory activities of the ethanolic extract of H. speciosa leaves (EEHS) against enzymes related to neurodegenerative diseases, inflammation, obesity, and diabetes were investigated. Furthermore, this study aimed at assessing the in vivo effects of the EEHS on the glycemia of normoglycemic and diabetic Wistar rats. Physicochemical characterization was performed by colorimetry and gas-liquid chromatography with flame ionization detection (GC-FID). The total number of colonies of aerobic mesophiles, molds, and yeasts was determined. The total coliforms and Escherichia coli were counted using the SimPlates kit, and sulphite-reducing Clostridium spores were quantified using the sulphite-polymyxin-sulfadiazine agar method. Salmonella spp. were detected using the 1-2 Test. The antioxidant activity of the EEHS was measured by its inhibition of 2,2'-azobis(2-amidinopropane) dihydrochloride- (AAPH-) induced oxidative hemolysis of human erythrocytes. The antimutagenic activity was determined using the Ames test. The acetylcholinesterase, butyrylcholinesterase, tyrosinase, hyaluronidase, lipase, α -amylase, and α -glycosidase enzyme-inhibiting activities were assessed and compared with commercial controls. The in vivo effects of the EEHS were assessed using the oral glucose tolerance test in normoglycemic Wistar rats and measuring the blood glucose levels in diabetic rats. The results demonstrated physical-chemical parameters of microbiological quality and safety in the leaves of H. speciosa , as well as antioxidant and antimutagenic activities and inhibition of enzymes related to neurodegenerative diseases, inflammation, obesity, and diabetes. In in vivo assays, it was shown that the normoglycemic rats challenged with glucose overload show significantly decreased blood glucose levels when treated with the EEHS. Taken together, the results ensure the microbiological quality and safety as well as showing the contents of carotenoids and polyunsaturated fatty acids of H. speciosa leaves. Additionally, the antioxidant, antimutagenic, anti-inflammatory, anti-Alzheimer's disease, anti-Parkinson's disease, antiobesity, and antihyperglycemic activities of the EEHS were demonstrated.