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The chestnuts are a group of nine species of deciduous trees and shrubs in the genus Castanea, in the beech family Fagaceae. Due to the nutritional properties, chestnuts have historically been used for human and animal consumption. Several by-products are produced during chestnut industrial processing, such as chestnut wood, flowers, leaves, shells, barks, and burs. Such by-products are a significant source of antioxidant compounds and can be used as food additives to be incorporated in food products to optimize nutritional and quality characteristics and to prevent oxidation processes. The aim of this research work was to optimize the conditions for the extraction of phenolic compounds from male chestnut flowers using ultrasonic-assisted extraction in order to develop an extract rich in phenolic compounds and, study it potential as a natural ingredient with potential industrial application. Time (t), ultrasound potencial (P) and solvent (S) were the conditions optimized by response surface methodology using a 5-level experimental design. The responses used as criteria were the quantification of the individual phenolic compounds identified by HPLC-DAD-ESI/MS and the extraction yield of the obtained residue. Based on their chromatographic, UV-vis and mass spectra characteristics, fourteen compounds were identified of which seven were hydrolysable tannins and seven were flavonoids being trigalloyl-HHDP-glucoside the major compound found. The conditions that maximized the total phenolic content was found at t= 23.47±2.90 min, P= 258.78±16.09 W and S= 50.51±7.11% ethanol producing an extract with 21.86±8.84 mg of phenolic compounds per g of extract. In this way, this study allowed to define the best conditions for the extraction of bioactive compounds and confirm the potential of the extract of male chestnut flowers as a natural ingredient for functional foods, providing health benefits.