Bone health is an important aspect of a good quality of life. Although bone-related diseases have low mortality rates, they can significantly impact people’s daily lives. This article aims at investigating the sensitivity of the detection of healthy and fractured bones using electrical impedance spectroscopy (EIS) through a 3D biological phantom. Two identical cylindrical containers were manufactured, and phantoms were created using a gelatin mixture and chilled turkey femurs. Electrode symmetry was assessed, and a calibration vector was established using a 0.9% NaCl solution. Impedance spectrum measurements were conducted for the phantoms with the gelatin mixture alone and with bones (intact and fractured at two distinct points) placed in the center of the container. Collected data showed equivalence, and it was only necessary to evaluate the electrode pairs 2–10, 3–11, and 4–12. Moreover, impedance module values were sufficient for data assessment, as the phase was equivalent in fracture evaluations and had a phase shift of 180 \(^\circ \) when evaluated at mirrored positions.Considering the voltage values in the electrode pair 2–10 on all three electrode layers an 86.29 % increase in the impedance module value was observed in layer 1, 70.29% in layer 2, and 43.94% in layer 3 for the phantoms with the intact bone compared to the phantoms with only the pure gelatin mixture. Furthermore, variations in impedance module values were observed when the fractured femur was added to the phantoms. The system demonstrated sensitivity to detecting intact bones and fractures at different electrode levels, which is a promising result for research involving EIS and bone fractures.