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"Guava" (Acca sellowiana) is an unconventional edible plant from Brazil. It is used in traditional medicine as an anti-diabetic; however, pharmacological studies on this plant are scarce. This study aimed to evaluate the chemical and safety profile of an aqueous A. sellowiana peel extract (ASPE) and its effects on endothelial EA.hy926 cells under glucose overload and in vivo (Artemia salina). An ethanolic extract from A. sellowiana peels (ASPE etOH) was also produced and characterized. Results showed that ASPE did not present in vivo toxicity, and it was found to contain high phenolic content and redox capacity. ASPE (50 µg/mL; 24 h) prevented oxidative stress and mitochondrial dysfunction, besides positively modulating Sirtuins 1 and 3, and prevented the increase of COX-2 and NF-kβ expression levels in EA.hy926 cells under glucose overload. Chromatographic fractionation, metabolite profiling, spectroscopic and bioinformatics analyses revealed the presence of phenolic acids, flavan-3-ols, flavonols, flavones, flavanones, and anthocyanidins, displaying a diversity of compounds in the crude and fractionated ASPE etOH. This study provided evidence on the safety profile, chemical composition, and pharmacological activities of A. sellowiana. [Display omitted] • Acca sellowiana "guava" is an Unconventional Food Plant from Brazil. • Extract from guava peels (GP) is a source of flavonoids and phenolic acids. • GP improved mitochondrial function in EA.hy92 cells under high glucose. • GP regulated SIRT 1, SIRT 3, COX-2, and NF-kβ endothelial expressions. • GP aqueous extract is in vivo safe. [ABSTRACT FROM AUTHOR]