A Two-stage Process for Increasing the Yield of Prebiotic-rich Extract from Pinus densiflora

The importance of polysaccharides is increasing globally due to their role as a significant source of dietary prebiotics in the human diet. In the present study, in order to maximize the yield of crude polysaccharides from Pinus densiflora, response surface methodology (RSM) was used to optimize a two-stage extraction process consisting of steam explosion and water extraction. Three independent main variables, namely, the severity factor (Ro) for the steam explosion process, the water extraction temperature ($^{\circ}C$), and the ratio of water to raw material (v/w), were studied with respect to prebiotic sugar content. A Box-Behnken design was created on the basis of the results of these single-factor tests. The experimental data were fitted to a second-order polynomial equation for multiple regression analysis and examined using the appropriate statistical methods. The data showed that both the severity factor (Ro) and the ratio of water to material (v/w) had significant effects on the prebiotic sugar content. The optimal conditions for the two-stage process were as follows: a severity factor (Ro) of 3.86, a water extraction temperature of $89.66^{\circ}C$, and a ratio of water to material (v/w) of 39.20. Under these conditions, the prebiotic sugar content in the extract was 332.45 mg/g.