Abstract: A new separation method by combining two TEVA extraction chromatographic columns was studied for analysis of trace neptunium in bulk uranium. The absorbed Np(Ⅳ) and residual uranium remaining on the first TEVA column will be eluted by ammonium carbonate after uranium matrix is removed with 3.0 mol/L HNO₃. After the carbonate complexation of Np(Ⅳ) ion in the elutant is destroyed with nitric acid, the sample is loaded on the second TEVA column, uranium may then be rinsed with 3.0 mol/L HNO₃ while neptunium remains on the TEVA resin. Then Np can be stripped out of the resin with 0.02 mol/L HF-0.02 mol/L HNO₃. The average recovery of neptunium is greater than 86% while the relative standard deviation is less than 4%(n=3), and the decontamination coefficient of uranium is more than 10⁵. This method has a good application prospect in separation neptunium from high U/Np mass ratio samples in nuclear fuel reprocessing plant.