The aim of the study was to investigate the effects of age and sex on the content of heavy metals (Pb, Cd, Zn and Cu) in the hair, liver and the longissimus lumborum muscle of roe deer (Capreolus capreolus L.). In addition, correlations between heavy metals in individual tested matrices were studied as well as an attempt was made to explain the reasons for their accumulation in specific research matrices. The levels of Zn and Cu were measured using atomic absorption spectroscopy (F-AAS). The levels of Pb and Cd were analysed by means of electrothermal atomic absorption spectroscopy (ET-AAS). In the own research, sex-related statistically significant differences in concentrations of analysed elements were confirmed. Higher levels of the elements were observed in samples collected from does (except for Cd in the liver samples of 6–7-month-old male fawns) but only in some cases, these values were statistically significantly higher. In some cases, age-related differences in the content of individual metals were also confirmed. Age-dependent increased Pb levels in the muscle and liver were reported in 6–7-month-old, <3–4-year-old, <5–6-year-old males. Similar relationships were confirmed for Zn in the bucks’ muscles. In the group of does, the increase in heavy metal concentrations with aging was observed for Zn in the muscle and for Pb in the liver. In the other cases, the trend of age-related level changes regarding heavy metals was not clearly demonstrated. Significant positive correlations between Cu and Cd were found for all samples, and significant negative correlations of Cu with Pb were found for the hair and liver. There was a negative correlation of Zn with Cu in all matrices. The study also revealed a negative correlation between Zn and Pb in muscle and a strong significant negative correlation between Zn and Cd in hair and in the liver.