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The main scope of this work is to develop nano-carbon-based mixed matrix celluloseacetate membranes (MMMs) for the potential use in both gas and liquid separation processes. Forthis purpose, a variety of mixed matrix membranes, consisting of cellulose acetate (CA) polymerand carbon nanotubes as additive material were prepared, characterized, and tested. Multi-walledcarbon nanotubes (MWCNTs) were used as filler material and diacetone alcohol (DAA) as solvent.The first main objective towards highly efficient composite membranes was the proper preparationof agglomerate-free MWCNTs dispersions. Rotor-stator system (RS) and ultrasonic sonotrode (USS)were used to achieve the nanofillers’ dispersion. In addition, the first results of the application of thethree-roll mill (TRM) technology in the filler dispersion achieved were promising. The filler material,MWCNTs, was characterized by scanning electron microscopy (SEM) and liquid nitrogen (LN2)adsorption-desorption isotherms at 77 K. The derivatives CA-based mixed matrix membranes werecharacterized by tensile strength and water contact angle measurements, impedance spectroscopy,gas permeability/selectivity measurements, and water permeability tests. The studied membranesprovide remarkable water permeation properties, 12–109 L/m2/h/bar, and also good separationfactors of carbon dioxide and helium separations. Specifically, a separation factor of 87 for 10%He/N2feed concentration and a selectivity value of 55.4 for 10% CO2/CH4feed concentrationwere achieved.