Poly (phenylene-ether-ether) sulfone membrane was fabricated and characterized by efficient glycerol/biodiesel separation produced from waste cooking oils trans-esterification. The membrane preparation was processed by phase inversion technique. The morphology, Physico-chemical properties and separation behavior of membranes were studied at various PPEES concentration. A uniform surface was observed for the prepared membranes by scanning electron microscopy. AFM images exhibited that surface roughness was decreased from 9.24 to 7.26 nm by the increase of PPEES concentration from 12 to 15 %wt. Similar trend was found for the membrane Flux, water content, and porosity by the increase of PPEES content ratio up to 15 %wt. The efficiency of glycerol removal and mechanical strength was also improved by the increase of polymeric matrix concentration.