The health care application of ionizing radiation has expanded worldwide during the last several decades. While the health impacts of ionizing radiation improved patient care, inaccurate handling of radiation technology is more prone to potential health risks. Therefore, the present study characterizes the bone dose response using bovine femurs from a slaughterhouse. The gamma irradiation was designed into low-doses (0.002, 0.004 and 0.007 kGy) and high-doses (1, 10, 15, 25, 35, 50 and 60 kGy), all samples received independent doses. The combination of FTIR spectroscopy and PLS-DA allows the detection of differences in the control group and the ionizing dose, as well as distinguishing between high and low radiation doses. In this way, our findings contribute to future studies of the dose response to track ionizing radiation effects on biological systems.Funda????o de Amparo ?? Pesquisa do Estado de S??o Paulo (FAPESP)Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior (CAPES)Conselho Nacional de Desenvolvimento Cient??fico e Tecnol??gico (CNPq)FAPESP: 17/50332-0; 21/00633-0CAPES: 001; PROCAD 88881.068505/2014-01CNPq: INCT-465763/2014-6; Sisfoton 440228/2021-2; PQ-309902/2017-7; PhD-grant-141946/2018-0; 142229/2019-9