Hexabromocyclododecane (HBCD) is a critical emerging brominated flame retardant to which consumers can be exposed at high doses through a single food intake. Based on an animal experiment involving 3 groups of laying hens fed during 70 days with a control diet or γ-HBCD-contaminated diets at 0.1 or 10 μg γ-HBCD g−1 feed, this study aims to use the volatolome of biological samples for revealing markers of livestock exposure to HBCD. Liquid chromatography–tandem mass spectrometry was used to monitor the time-course of HBCD levels in bodily samples. Each liver was analyzed by solid-phase microextraction–gas chromatography–mass spectrometry for volatolome profiling. After 70 days, γ-HBCD concentrations in egg yolk, fat, liver and serum reached 54 ± 4, 85 ± 6, 31 ± 6, and 32 ± 4 ng g−1 lw, respectively, for the low exposure level and 4.6+/5.7, 7.8+/6.5, 3.9+/3.0 and 3.9+/6.1 μg g−1 lw, respectively, for the high exposure level. Isomerization of γ-HBCD into α- and β-HBCD was observed in all tissues, at least for the high exposure level. Volatolome data allowed a significant discrimination between control and exposed animals whatever the feed contamination load, demonstrating a liver metabolic response to γ-HBCD exposure. The relevance of the twenty nine volatile exposure markers tentatively identified was discussed in light of literature data.