Shuttle of polysulfide from the sulfur cathode to lithium metal anode in rechargeable lithium–sulfur batteries is a critical issue hindering cycling efficiency and life. Several approaches have been developed to minimize it including polysulfide-blocking separators; there is a need for measuring polysulfide transport through separators. We here show a linear voltage sweep method to measure anodic (oxidization) current of polysulfide crossed separators, which can be used as a quantitative measurement of the polysulfide transport. The electrochemical oxidation of polysulfide is diffusion controlled. The electrical charge in Coulombs produced by the oxidation of polysulfide is linearly related to the concentration of polysulfide within a certain range (≤0.5 M). Separators with a high porosity (large pore size) show high anodic currents, resulting in fast capacity degradation and low Coulombic efficiencies in Li–S cells. These results demonstrate this method can be used to correlate the polysulfide transport through separators with the separator structure and battery performance, therefore provide guidance for developing new separators for lithium–sulfur batteries.