The soil is a very complex and dynamic system and its contamination has become one of the main environmental concerns. Sugar-ethanol mills, for example, have a high hazard potential, mainly soil contamination, due to the production of different residues. The use of vinasse, a by-product of ethanol production, as fertilizer is a technically and economically feasible alternative. However, excessive quantities have been used, causing adverse changes in the soil. The present study was aimed at evaluating the toxic potential of vinasse in the Tradescantia pallida test system using the micronucleus test (Trad-MCN). Ten plants were exposed to ultrapure water as the negative control (NC), in MMS as the positive control (PC), raw vinasse (RV), and vinasse diluted to 50% (C1), vinasse diluted to 25% (C2), vinasse diluted to 12.5% (C3). The medium buds of each plant were used to assess the mutagenic effects based on the presence of micronuclei in tetrads. The results revealed that RV and C1 had a mutagenic potential compared to that of NC. The alterations observed suggest damage to the DNA of the test organism, which may not be reintegrated to the cell nucleus. The findings suggest caution in the application of this product to the soil despite this being an economically viable practice.