The extraction of uranium(VI) from sulfuric acid medium with tri-octylphosphine oxide (TOPO) in n-heptane was studied. Accompanied with the increase in the concentration of H2SO4, the distribution coefficient of uranium(VI) increased in the region of dilute sulfuric acid. When the concentration of H2SO4 surpassed 3.5 mol·dm−3, the distribution coefficient of uranium(VI) was at maximum. This result was due to the competition extraction between uranium(VI) and H2SO4. From the data, the composition of extracted species and the equilibrium constant of extraction reaction have been evaluated, which were (TOPOH)2UO2(SO4)2 (TOPO) and 107.6±0.15, respectively.