Response surface methodology along with Box-Behnken design was firstly applied to optimize the extraction conditions for the polysaccharides from the roots of Isatis tinctoria L. The experimental data obtained were fitted to a second-order polynomial equation using multiple regression analysis. The 3-D response surface and the contour plots derived from the mathematical models were applied to determine the optimal conditions. The optimal conditions were extraction temperature (X1) 99.5 °C, extraction time (X 2) 3.75 h and ratio of water to raw...