The objective of the present study was to optimise the conditions for extraction of bioactive compounds from drumstick (Moringa oleifera) seeds to determine their potential as a source of nutraceuticals. A central composite face centred design was applied to determine the effects of ethanol concentration (%), extraction temperature (°C), extraction time (min) and particle size (mm) on total phenolic content, total flavonoid content, total antioxidant activity, ferric reducing power and hydrogen peroxide scavenging activity. A second-order polynomial model was used for predicting the responses. The optimum extraction conditions were found to be ethanol concentration, 49.8 %, extraction temperature, 80 °C, extraction time, 45 min and particle size, 0.62 mm. LC–MS analysis under the optimum conditions led to the identification of 12 bioactive compounds. Major compounds were identified as 1H-pyrazol-4-yl methanol, 1-(2-methyl-5-nitro-1H-imidazol-1-yl)propan-2-ol, 1,4-naphthoquinone, luteolin-6-c-glucoside and hesperidin, which possess different biological activities.