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Kudingcha (KDC) is an important tea substitute containing abundant antioxidants. Herein, a ultrasonic-assisted extraction (UAE) technique based on deep eutectic solvents (DESs) was applied to optimize the total phenolic/total flavonoid content (TPC/TFC) from the KDC extracts. Results indicated that DES composed of L-proline and glycerol (Pro-Gly) had excellent extraction performance for TPC, TFC, ABTS^•+ and FRAP, which were significantly better than other solvents. Response surface methodology (RSM) was used to obtain optimal extraction parameters for simultaneously maximizing the TPC, TFC and antioxidant activity. Results revealed that water content in Pro-Gly, liquid to solid ratio (L/S), ultrasonic temperature and extraction time were the major influence factors of the TPC, TFC, ABTS^•+ and FRAP of the KDC extracts. The optimal conditions included water content in Pro-Gly of 46.4%, L/S of 25:1 (mL/g), ultrasonic temperature of 55 °C and extraction time of 50 min. Meanwhile, HPLC-MS/MS was adopted to identify the KDC extracts, which revealed the presence of major phytochemicals, including 5-chlorogenic acid, 4,5-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, kaempferol 3-rutinoside, myricetin and isorhamnetin. Moreover, UAE–Pro-Gly achieved further higher individual phenolics contents, TPC, TFC, ABTS^•+ and FRAP than other methods. In conclusion, UAE–Pro-Gly is a highly efficient method for extraction of phenolic antioxidants from KDC. Full article