The performance of vanadium redox flow batteries (VRFBs) is impacted by the diffusion and migration of the vanadium species across the separator. In this work, the vanadium crossover as a function of current density for vanadium-containing electrolytes of various state of charge (SOC) and state of discharge (SOD) is measured. Experiments conducted with electrolytes at complete charge/discharge yielded direct measurements of the transference numbers of the vanadium species. The transference numbers of V2+, V3+, VO2+, and VO2
+ were estimated to be 0.064 ± 0.002, 0.087 ± 0.003, 0.068 ± 0.003, and 0.018 ± 0.002, respectively. Experiments conducted with electrolytes at intermediate states of charge/discharge yielded direct measurements of the sum of transport numbers of the vanadium species. The transport number estimates are quantitatively related to faradaic efficiency loss and capacity fade of a working VRFB.