Glyphosate's remarkable effectiveness in weed control has made it among the foremost widely employed herbicides globally. Several Glyphosate-Based Herbicides (GBHs) contain 1-4 dioxane, a compound known to induce cancer in animals and potentially contribute to liver and kidney damage in humans. The objective was to assess the liver and kidney toxicity resulting from sub-chronic exposure to GBH in rats. Eighty wistar rats were divided into 8 groups, each consisting of 5 males and 5 females. These groups were categorized into 4 inhalation exposure groups and 4 oral exposure groups. The control groups were subjected to exposure to sodium chloride solution, while the other groups were exposed to GBH at low, medium, and high concentrations. This exposure continued for 75 days. Blood samples were taken for the assessment of Aspartate Aminotransferase (AST), Alanine Aminotransferase (ALT), creatinine, and urea levels, and the liver, kidney, pancreas and spleen tissues were sampled for histopathology. The females in the high oral concentration group exhibited the most elevated levels of AST, ALT, and urea (p<0.05). Liver steatosis was observed in all animals exposed to medium and high GBH concentrations (p<0.05). Tubular changes were evident in all GBH-exposed animals. Furthermore, animals submitted to a high GBH concentration displayed an increased count of nucleoli-organizing regions in the liver and kidney (p<0.05). High-concentration exposure to GBH, mainly orally, causes greater liver and kidney damage. Females had more pronounced hepatic and renal biochemical changes than males.