Phenolic compounds of Morus alba leaves are bioactive compounds with beneficial properties for human health. Therefore, in this study, an optimization of ultrasonic assisted extraction by Box-Behnken design was used for the first time to optimize factors such as the percentage of ethanol, ratio solvent/sample (v/w) and extraction time to reach the highest phenolic compound amounts (evaluated by HPLC-MS) while also evaluating in vitro antioxidant activity using DPPH, ABTS and FRAP assays. The optimal extraction conditions were 40% ethanol, 1/400 (w/v) and 35 min. Applying these optimal conditions, which were identified and quantified by HPLC-MS, resulted in the extraction of 21 phenolic compounds. According to these results, the main phenolic compounds in Morus alba leaves are the phenolic glycoside and phenolic acid named protocatechuic acid-glucoside and caffeoylquinic. In addition, Morus alba leaf extract contains flavonols such quercetin-3-O-6-acetylglucoside and rutin, which represent more than 7% of its total phenolic content.
Keywords: Box-Behnken; HPLC-MS; Morus alba leaves; antioxidant activity; phenolic compounds.