Curcuma aeruginosa is the common name of temu hitam in Indonesia, and the rhizome parts of the plant have several pharmacological activities. Generally, pharmacological activities are associated with bioactive content in the extract of medicinal plants. Several factors can in luence the bioactive extraction from medicinal plants such as solvent types, extraction time, extraction technique, and liquid-to-solid ratio. In this research, the extraction factors for extraction yield and cytotoxic activity of C. aeruginosa rhizome were optimized using the Box-Behnken experimental design. Effect of ethanol concentration, the ratio of liquid to solid, and extraction time for the maceration process was studied. The cytotoxic activity was determined by the brine shrimp lethality test. The optimum value that maximizes the extraction yield was 70% ethanol, 300:15 ml/g liquid to solid ratio, and 1-day extraction time. The optimum value that maximizes the cytotoxic activity was 70% ethanol, 150:15 ml/g liquid to solid ratio, and 2-day extraction time. The predicted extraction yield and cytotoxic activity at these projected values are 14.78% and 78.26 mg/l, respectively. In this model, Adeq Precision (10.35 and 4.16), R-Squared (0.86 and 0.79), and F-value (7.92 and 2.04) is rational to it the model for extraction yield and cytotoxic activity from C. aeruginosa rhizome.