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As a typical hydrogen storage material, titanium has an important application on hydrogen energy materials and deuterium-tritium fusion neutron source. The various contents and distribution of hydrogen isotopes in titanium will affect the structure and properties of titanium in many ways. The conventional quantitative determination methods of hydrogen isotope content in titanium include pressure-volume method, weighing method and thermogravimetric method, etc. Compared with these conventional methods, the thermal desorption spectroscopy (TDS) method can not only obtain hydrogen isotope contents, but also distinguish between hydrogen and deuterium, and can reveal the position information of hydrogen isotope atoms in the crystal lattice. In this research, by optimizing the hydrogen absorption process of titanium, titanium hydrogenated/deuteride samples with different hydrogen and deuterium contents were successfully prepared. Combined with X-ray diffraction (XRD) analysis, the crystal structure changes of titanium before and after hydrogen absorption are given. Using the TDS technology developed by our research group, the ion current signal intensity of the mass spectrometer was calibrated through a standard leakage, and the quantitative analysis of the amount of hydrogen isotope desorption in titanium was realized. The absolute error between the TDS measurement result and the pressure-volume method measurement value is less than 6%. The total contents of hydrogen and deuterium desorbed in the sample were calculated. The results provide a useful reference for the accurate measurement of hydrogen isotope content in materials in the field of hydrogen energy, nuclear energy and nuclear technology.