In this study, response surface methodology was used to study the effects of H 2 SO 4 , concentrations, substrate loading and residence time on liberation of reducing sugars (RS), total sugars (TS) and total phenolic compounds from poplar leaves and twigs. Box-Behnken design with three variables and three levels showed maximum release of total phenolic compounds (57.39 mg/ml) corresponding to 0.8% H 2 SO 4 concentration, 15% substrate level with residence time of 4 h. Under these conditions, the TS and RS released up to 161.20 and 5.24 mg/ml, respectively. Analysis of the pretreated substrate by Fourier transform infra-red and X-ray diffraction revealed the effectiveness of pretreatment conditions. Second-order polynomial equation using analysis of variance was employed for analyzing the results. The proposed model was found very significant with F-value of 48.39. The R 2 and adjusted R 2 values also revealed the accuracy of predicted response.
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