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Symmetrical Isotropic macroporous (SIM) membranes are one of the most desirable membrane morphologies with regards to microfiltration applications, namely for the disinfection of water and wastewater.In this study, Biodegradable Chitosan-Glycerol SIM membranes, prepared in acetic acid via a novel process for enhanced controlled architecture, suitable for wastewater microfiltration applications were biosynthesized.The macroporous isotropic cross-section of the membranes increased as the glycerol content increased, corresponding with a decrease in density from 38.96μg/mm3 to 9.706 μg/mm3.The membrane density was used to determine the effect of Glycerol to maintain the macroporous morphology of the Chitosan-Glycerol membrane matrices and the successful displacement of the chito-acetate complex within the membrane matrix.