Moringa oleifera is considered one of the most useful plants in the world because it's rich in bioactive substances, which employing on modern medical treatment, also can be used for many fields: pharmaceutical, food and cosmetics purposes. In this study, the response surface methodology (RSM) based on a Box–Behnken design (BBD) was employed to optimize the extraction time (X1: 20–60 min), extraction temperature (X2: 15–45 °C) and solvent-solid ratio (X3: 5–7 ml/g), to obtain a high crude of flavonoids yield from Moringa oleifera Leaves by ultrasonic-assisted extraction technique (UAE). The optimum conditions were an extraction time 23 (min), extraction temperature 44 (°C) and solvent-solid ratio 5.05 (ml/g). Under these conditions, the experimental yield was 72.65 (mg ER/g), well matched with the predicted yield 74.34 (mg ER/g) with the coefficients of determination (R2= 0.9861), thus indicating the suitability of response surface methodology in optimizing the ultrasound-assisted extraction of flavonoids from Moringa oleifera Leaves.