Response surface methodology was used to optimize experimental conditions for ultrasound-assisted extraction of functional components from Medicago sativa L. (Fabaceae) aerial parts. The procedure was performed according to the Box-Behnken design. The independent variables were ethanol concentration (X1), temperature (X2), and pH (X3), while the responses were extraction yield, DPPH radical scavenging activity of the extracts (RSA IC50), content of total phenols (TP), total flavonoids (TF) and total phenolic acids (TPA). The response values of the extracts prepared at optimum conditions (X1, X2, X3) were: yield (162 mg/ml, 20 %, 60 °C, 5.5), TP (6.63 mg/ml, 100 %, 60 °C, 5.5), TF (0.292 mg/ml, 100 %, 23 °C, 5.5), TPA (0.275 mg/ml, 100 %, 23 °C, 8.5) and RSA IC50 (3.97 mg/ml, 40 %, 60 °C, 7.0). The predicted values matched well with the observed ones indicating the suitability of the selected models for extraction of bioactive compounds from M. sativa.