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Free keywords:
Mural binders, Response surface method, Protein extraction, GuHCl solution
Abstract:
In this paper, response surface method (RSM) was used to study the effect of extraction parameters on the extracting efficiency of guanidine hydrochloride (GuHCl) on residual protein from mural model samples. The central composite design (CCD) was applied to find the optimal combination of variables, including GuHCl concentration (x: 1.5-2.5 M), extraction time (y: 90-180 min), temperature (z: 50-70 °C) and ultrasound power (α: 120-280 W) during the extraction. Multiple regression analysis was employed to fit the experimental data into a second-order polynomial equation, and appropriate statistical methods were used for analysis. The optimal extraction conditions were determined as follows: GuHCl concentration of 1.89 M, extraction time of 132 min, temperature of 57 °C, and ultrasonic power of 210 W. The extraction efficiency was 16.685 % under the optimal conditions, which was in agreement with the theoretically predictive value of 17.707 %. The extraction process under optimized parameters had no significant impact on the collagenous structures, characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), ultraviolet (UV) absorption spectroscopy and circular dichroism (CD) spectroscopy.
