The contribution is aimed at the development of methodology that allows to consider green analytical chemistry criteria during optimization of liquid chromatographic separation with design of experiment. The objectives of the optimization are maximization of peak areas of five non-steroid anti-inflammatory drugs, maximization of resolution between peaks, with simultaneous shortening of chromatographic separation time and minimization of mobile phase environmental impact. This is obtained with design of experiment to consider many experimental conditions and Derringer's desirability function to combine many optimization objectives. The possibilities of introduction different green analytical chemistry metrics are discussed and the methodology of mobile phase greenness assessment is proposed. The optimal response for all objectives is obtained for 0.96 mL min(-1) of mobile phase flow rate, 61% of MeOH content, temperature of 25 . 0 and pH equal to 4.5. The separation takes less than 9 min.