Black nighstshade (Solanum nigrum L.) is a plant proven to be beneficial towards human health in which one of it is its potential for antioxidant activity. In this research, the optimization of blanching temperature and time on antioxidant activity of black nightshade fruit was observed. The fruit was extracted using 80 % ethanol food grade and by microwave assisted extraction with different temperature ranged from 70 °C up to 90 °C and different time ranged from 3 up to 10 minutes. The combination was calculated using Central Composite Design by Response Surface Methodology (RSM). The effect of different blanching temperature and time conditions on the yield, total phenolic content, total flavonoid content and antioxidant activity using DPPH radical scavenging assay of the black nightshade fruit were observed. Blanching temperature and time have no significant effects on total phenolic and total flavonoid compound. There was a siginificant effect on antioxidant activity, therefore the optimization was able to be obtained by RSM. The highest antioxidant activity of black nightshade fruit was obtained at 80.4 °C for 3.3 minutes in which the predicted result was 87.82 ppm while the challenge test's result was 88.10 ppm. It showed 99.6 % accuracy therefore the optimization's model was suitble. The yield, phenolic and flavonoid content of black nightshade fruit were also known to have negative correlation with its antioxidant activity.