Blueberry residue is usually discarded as waste,but has a high anthocyanins content. The extraction methodof anthocyanins from blueberry residue with ultrasonicassisted dual-aqueous phase system was optimized. Interms of the principle of central group and design (CCD)experimental design, three-factor and five-level responsesurface analysis was adopted to optimize the extractionconditions with the extraction rate of anthocyanins. Theoptimum extraction rate of anthocyanin was12.372 ± 0.078 mg/g. Anthocyanin extract could protectthe pBR322 DNA oxidative damage induced by Fentonreagent, increase the superoxide dismutase(SOD) and glutathioneperoxidase (GSH-Px) enzyme activities, anddecrease the H2O2-induced cell apoptosis of human normalliver cell (LO2 cell). The study indicates that the extractionrate of anthocyanin was increased by optimized ultrasonicassisted dual-aqueous phase system. The anthocyaninextract could protect DNA and LO2 cell from oxidativedamage.