This study aimed to evaluate the presence of bioactive antioxidant molecules in chestnut outer shells. For this proposal, several extraction procedures were used, specifically different solvents (water, ethanol:water (80:20) and acetone:water (80:20)) combined with two extraction methods (stirring and ultrasounds (US)) and times of extraction. Several chemical assays were applied to evaluate the flavonoids, hydrolysable and condensed tannins contents, and the antioxidant activity by total reducing capacity, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and reducing power. Some individual compounds were identified by HPLC-UV detection. Furthermore, the antimicrobial activity of the obtained extracts was screened using different microorganisms, namely Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Enterococcus faecalis) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Proteus mirabilis, Klebsiella pneumoniae), as well as fungi (Candida albicans). The mixture acetone:water (80:20) was the best solvent to extract condensed tannins and flavonoids. It was with this mixture that the best total reducing capacity, (DPPH) radical scavenging activity and reducing power were determined. The highest extraction yield was also obtained with this mixture. Only for the hydrolysable tannins, the most suitable extraction solution was ethanol:water (80:20). The presence of gallic and tannic acids was detected by HPLC-UV. Concerning the extraction methods, only the extracts obtained by the US exhibited antimicrobial activity. The ethanolic extract showed antimicrobial activity against the Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Enterococcus faecalis) and the extracts with acetone:water (80:20) and water showed antimicrobial activity against Proteus mirabilis. The results of this study demonstrated that the chestnut outer shell is a promising source of bioactive compounds. FCT, Portugal [CIMO: UID/AGR/00690/2019]; Portugal 2020 [Go_ClimCast Project: PDR2020- 1.0.1-FEADER-032060] info:eu-repo/semantics/publishedVersion