Positron annihilation spectroscopy is applied to high‐quality β‐Ga 2 O 3 [010] oriented 5 μm thick epilayer before and after electron irradiation to calculate absolute vacancy concentrations. Room temperature irradiated is carried out with 2.5 MeV electrons to a dose of 1.9 × 10 19 cm −2 to generate high concentrations of vacancies. Both oxygen and gallium vacancies are generated. The vacancy‐sensitive depth‐dependent S ‐parameter and the positron diffusion length were measured to determine vacancy concentrations. The positron diffusion length dropped from 101 ± 2 nm to 47 ± 2 nm while the S ‐value in the epilayer rose 2.1 ± 0.3%. The positron annihilation spectroscopies (PAS) results on the irradiated film, in comparison with unirradiated as‐grown high‐quality bulk single crystals, are used to obtain for the first time a quantitative estimate of the defect concentration from the S ‐parameter for various samples. The −3 charge state of gallium vacancies is taken into account. PAS results on irradiated silicon are shown for contrast, to illustrate the complications in β‐Ga 2 O 3 introduced by a significant orientation dependence and, likely, the presence of hydrogen.