Platelet-rich fibrin (PRF) is widely used blood-derived biomaterial which is directly applied to the surgical wounds. Depending on its autologous origin, PRF is thought as a safe material. However, it is not known to what extent the blood-derived toxins can be found in the PRF by considering the systemic exposure rates of the individuals to the toxins. The aim of this pilot study was to test the hypothesis whether PRF contains any blood-origin heavy metals (HMs) and smoking increases their concentrations as an environmental HM source. PRF samples were obtained from systemically healthy 30 non-smoker and 30 smoker volunteers. All liquid and dry fibrin parts of the PRF samples were analyzed in terms of 15 toxic elements using inductively coupled plasma mass spectrometry. All analyzed HMs were detected in all investigated PRF samples within various concentrations in both groups. In addition, significantly high levels of cadmium, arsenic, lead, manganese, nickel, chromium, and vanadium were detected in dry fibrin matrices of PRF samples of smokers comparing with non-smokers (p < 0.05). Only cadmium was at significantly high levels in the liquid part of PRF samples of smokers (p < 0.05). This is the first study evaluating toxic ingredients of PRF. The results revealed that PRF contains various toxic HMs. Additionally, systemic exposure to environmental HM sources such as smoking may significantly increase HM concentrations in PRF. Further studies are required to investigate the transmission potentials of HMs to the applied tissues and biological importance of PRF-origin HMs.