Abstract: BIXS (β-ray induced X-ray spectrometry) is a nondestructive method to analyze tritium, which can be affected by many factors in inversion process. The difference between tritium film’s thickness and inversion basic function spectrum’s thickness can’t be overlooked. By the Monte Carlo method code PENELOPE and experiment, it is found that the thickness error of accelerator tritium film has an effect for inversion of BIXS method. The thickness error makes a great impact on tritium increasing distribution, and a less impact on tritium descending distribution. The bigger the slope of tritium depth distribution in titanium film, the greater the influence of thickness error of tritium film on BIXS inversion.