Background/Aims: Carbon Black (CB) is the most widely produced and commercially used nanocarbon. Growing evidence links nanomaterials to adverse effects, arising from their large surface area capable of interacting with biological systems. Due to the variety of applications and human exposures to nanoparticles, it is important to assess the potential health risk and understand the underlying mechanisms of their toxicity. The present study aims to investigate the cytotoxic effect of the Carbon black nanoparticle covalently linked to ethylenediamine (CB-EDA) on AML-12 cells, a lineage of murine hepatocytes. Methods: For this, the cells were exposed to CB-EDA for 24h, at different concentrations of the nanoparticle (1, 50, 250, 500 and 1000 μg/mL). Effects on cell viability were evaluated using MTT and neutral red dye tests and analysis of changes in cell morphology. Furthermore, the oxidative profile (levels of reactive oxygen species - ROS and nitrogen - RNS) and inflammatory profile (production of IL-6 and TNF-α) were investigated. Results: The results show that CB-EDA nanoparticle causes a reduction in cell viability at concentrations of 250, 500 and 1000 μg/mL after the exposure period. There was a significant increase in the production of ROS, RNS and pro-inflammatory interleukin TNF-α. Conclusion: The data suggest that CB-EDA nanoparticle has a cytotoxic potential in AML-12 hepatocytes, evidenced by the induction of oxidative stress and secretion of inflammatory cytokines, with a consequent decrease in cell viability.