Box-Behnken design-based optimization for deproteinization of crude polysaccharides in Lycium barbarum berry residue using the Sevag method

Deproteinization of crude polysaccharides in the residue from Lycium barbarum berries (LBBs) was conducted using the Sevag method. A Box-Behnken design based on single-factor experiments was employed to optimize the deproteinization technology. The results showed that the deproteinization conditions had significant effects on the extraction yield of polysaccharides and the residual protein content in Lycium barbarum polysaccharides (LBP). The experimental data were fitted to a second-order polynomial equation, using multiple regression analysis with a high coefficient of determination (R²) value. The optimal conditions were as follows: a ratio of raw material to water extract concentrate from the residual LBBs of 0.15 g/mL; a ratio of chloroform to n-butyl alcohol of 2.17 mL/mL; and a ratio of water extract concentrate from residual LBBs to Sevag reagent of 0.50 mL/mL; with a maximum polysaccharide yield of 0.49% and minimum residual protein content of 0.087%. The results were confirmed through validation experiments. GPC analysis indicated that deproteinized LBP molecules became much more homogeneous. X-ray diffraction indicated that the hydrogen bonding of deproteinized LBP was weakened. This optimization of LBP should be a useful method for purification of crude LBP.