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Food safety can be improved using ionizing radiation to reduce food spoilage and to extend its shelf life. The gas chromatography/mass spectrometry (GC/MS) has been validated by the European Community as a powerful method to identify irradiated food containing fat. The preliminary goals of our research were: (i) to set up this method, based on the detection of radiation induced 2-dodecylcyclobutanones (2-DCB) in pork muscle samples and (ii) to check the microbiological efficacy of the treatment. The main objective was to render the GC/MS a quantitative technique for dose estimation, through the measurement of the 2-DCB concentration in the irradiated sample. Our results show that the reduction of the microbial population is substantially reduced even at 2 kGy, and that a clear identification of irradiated samples can be achieved also one month after irradiation at 2 kGy in frozen-stored samples. The 2-DCB concentration showed a linear dependence on dose in the range 1−10 kGy, no matter the origin of the sample; a unique calibration function was obtained, that allowed dose estimation in irradiated pork samples. A retrospective evaluation on the quality of the treatment could be carried out this way.