Pendimethalin and trifluralin are widely used dinitroaniline herbicides. Both compounds can be found as residue levels in agricultural products. This study was conducted in order to provide necessary information for the risk assessment of pendimethalin and trifluralin. In this study, reactive oxygen species (ROS) levels were measured to examine the potential of both compounds to induce oxidative damage in Chinese hamster lung fibroblast (V79) cells. Also, the genotoxic effects of pendimethalin and trifluralin at the concentration range of 1-500 μM was determined. Single cell gel electrophoresis (comet) and micronucleus assays were used on human peripheral lymphocytes and V79 cells for the genotoxicity assessment. The cell viability of two dinitroaniline herbicides were determined by the use of neutral red uptake assay on V79 cells. IC50 values were determined as 66 μM and 128 μM for pendimethalin and trifluralin, respectively. They significantly increased ROS levels on V79 cells for 1-24 h. Both herbicides significantly induced the DNA damage and showed genotoxicity on lymphocytes and V79 cells. Micronucleus frequency increased significantly after pendimethalin and trifluralin treatment of the lymphocytes and V79 cells. Therefore, we concluded that both of the herbicides induced the genotoxicity through the activation of oxidative stress pathway and chromosomal damage.