Abstract: The extraction effects of six deep eutectic solvents on burdock polyphenols (BPs) were investigated, and the response surface methodology was used to optimize the process conditions of ultrasonic-assisted deep eutectic solvent extraction of BPs, moreover, the total reducing power, the DPPH• scavenging capacity, the inhibition of α-glucosidase and cholic acid binding capacity in vitro were studied, and its composition were analyzed by HPLC. The results showed that DES 5 (the molar ratio of choline chloride to ethylene glycol was 1:4) was the best solvent system for the extraction of BPs. The optimum technological conditions were as follows, the water content of deep eutectic solvent was 40%, the ratio of material to liquid was 55:1 mg/mL, the extraction temperature was 49 ℃, and the extraction time was 41 min, under the above conditions, the yield of BPs was 1.19%±0.11%. The biological activity analysis showed that the IC₅₀ of the total reducing power and DPPH• scavenging capacity of burdock polyphenol were 0.93 mg/mL and 7.03 μg/mL, respectively, indicating that it had good antioxidant capacity, but weaker than V_C. The IC₅₀ of α-glucosidase inhibition was 0.13 mg/mL, which was higher than acarbose, indicating that it possessed excellent hypoglycemic activity. In addition, its cholic acid binding rate was 33.23% when the mass concentration was 1.5 mg/mL, implied that it had a certain hypolipidemic activity. HPLC analysis showed that BPs contained chlorogenic acid, caffeic acid and ferulic acid. The results would provide a theoretical basis for the green preparation of burdock polyphenol and its development and utilization as a functional component of antioxidation, hypoglycemia and blood lipids.