The biological effects of tritium in humans need to be clarified, because the chances of humans becoming exposed to tritium beta radiation may increase with the development of the nuclear fusion reactor. To evaluate the biological effects of tritium, it is necessary to estimate exactly the absorbed dose from the tritium beta rays in the tissue. In many reports, the absorbed dose of HTO in the tissues is estimated from the tritium content in body fluid and dose calculations are customarily based upon the water content of soft tissues, which is taken to be 0.7 to 0.8. However, these methods may not show the exact absorbed dose in the organs. In the present study, the radioactivity of the critical tissues was measured directly using a sample oxidizer and the absorbed dose was calculated from the radioactivity of tritium in the tissues. Details on the method for calculation of the absorbed dose in tissues of the mouse is shown in this report. The results suggest that the absorbed dose should be obtained from the radioactivity in the tissues.