Extraction of rutin from Ruta graveolens L. leaves was performed in choline chloride based deep eutectic solvents (DESs). Firstly, rutin extraction was performed in five different DESs; pure DESs and with different amounts of water added (25% and 50% v/v), for different times (1 h, 3 h and 6 h) in order to find the best DES for further optimization. Choline chloride plus citric acid (1 : 1) was shown to be best DES and optimization of rutin extraction was performed using response surface methodology (RSM) based on the selected solvent and applying different extraction parameters: extraction time, temperature and water content. The rutin content was determined by HPLC and varied from 0.55 g/100 g of plant to 1.81 g/100 g of plant depending on the applied conditions. The best conditions for rutin extraction were found to be: a temperature of 70°C, an extraction time of 52 min and a water content of 27 % for the extraction of 1.88 g /100 g plant.