Extraction optimization of pepsin-soluble collagen from eggshell membrane by response surface methodology (RSM)

Response surface methodology (RSM) was used to investigate the effect of extraction-process variables on pepsin-soluble collagen (PSC) from eggshell membrane. A central composite design (CCD) was employed for experimental design and analysis of the results to obtain the best possible combination of NaOH concentration (X1: 0.4-1.2 mol/l), alkali treatment time (X2: 6-30 h), enzyme concentration (X3: 15-75 U/mg) and hydrolysis time (X4: 12-60 h) for maximum PSC extraction. The experimental data obtained were fitted to a second-order polynomial equation using multiple regression analysis and analyzed by appropriate statistical methods. According to the results, optimum extraction conditions were as follows: NaOH concentration of 0.76 mol/l, alkali treatment time of 18 h, enzyme concentration of 50 U/mg and hydrolysis time of 43.42 h. The experimental extraction yield under optimal conditions was found to be 30.049%, which is in good agreement with the predicted value of 30.054%.