Olive tree pruning biomass (OTP) and olive mill leaves (OML) are the main residual lignocellulosic biomasses that are generated from olive trees. They have been proposed as a source of value-added compounds and biofuels within the biorefinery concept. In this work, the optimization of an ultrasound-assisted extraction (UAE) process was performed to extract antioxidant compounds present in OTP and OML. The effect of the three parameters, ethanol/water ratio (20, 50, 80% of ethanol concentration), amplitude percentage (30, 50, 70%) and ultrasonication time (5, 10, 15 min), on the responses of total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activities (DPPH, ABTS and FRAP) were evaluated following a Box-Behnken experimental design. The optimal conditions obtained from the model, taking into account simultaneously the five responses, were quite similar for OTP and OML, with 70% amplitude and 15 min for both biomasses and a slight difference in the optimum concentration of ethanol. (54.5% versus 51.3% for OTP and OML, respectively). When comparing the antioxidant activities obtained with OTP and OML, higher values were obtained for OML (around 40% more than for OTP). The antioxidant activities reached experimentally under the optimized conditions were 31.6 mg of TE/g of OTP and 42.5 mg of TE/g of OML with the DPPH method, 66.5 mg of TE/g of OTP and 95.9 mg of TE/g of OML with the ABTS method, and 36.4 mg of TE/g of OTP and 49.7 mg of TE/g of OML with the FRAP method. Both OTP and OML could be a potential source of natural antioxidants.