Aiming at the self-excitation effect on the interference of measurements which exist in the process of Energy dispersive X-ray fluorescence method for uranium measurement. To solve the problem of radioactive isotopes only used as excitation source in determination of uranium. Utilizing the micro X-ray tube to test Self-excitation effect to get a comparison of the results obtained by three different uranium ore samples—109Cd, 　241Am and Mirco X-ray tube. The results showed that self-excitation effect produced the area measure of characteristic X-ray peak is less than 1% of active condition, also the interference of measurements can be negligible. Photoelectric effect cross-section excited by 109Cd is higher, corresponding fluorescence yield is higher than excited by 241Am as well due to characteristics X-ray energy of 109Cd, 22.11 & 24.95 KeV adjacent to absorption edge energy of Lα, 21.75 KeV, based on the above, excitation efficiency by 109Cd is higher than 241Am; The fact that measurement error excited by 241Am is significantly greater than by 109Cd is mainly due to peak region overlap between L energy peaks of uranium and Scattering peak of 241Am, 26.35 keV, These factors above caused the background of measured Spectrum higher; The error between the uranium content in ore samples which the X-ray tube as the excitation source and the chemical analysis results is within 10%. Conclusion: This paper come to the conclusion that the technical quality of uranium measurement used X-ray tube as excitation source is superior to that in radioactive source excitation mode.