The objective of this research was to assess the impact of 2-methyl-2-nitro-3-guanidine on mammalian species, otherwise not the intended targets, followed by one-time exposure and its adverse effects. Thirty-five Sprague Dawley rats of either male or female sex, aged four weeks, and with an average weight of 80±20g, were divided into three groups randomly: control E exposed and exposed and supplemented with vitamin C, and assigned the titles C,E, and V, respectively. Each group comprised fifteen rats. Both E and V groups were further divided into subgroups: E1, E2, and E3, exposed to LD10, LD25, and LD50 doses of Dinotefuran respectively, and V1, V2, and V3, exposed in the same manner but supplemented with a fixed dose of ascorbic acid (vitamin C). Ascorbic acid was administered in aqueous form 35mg/100mL of water and provided ad libitum. Eight hours after exposure to Dinotefuran, 5ml of blood was collected under sedation through the cardiac vein. After 48 hours, two rats randomly selected from each subgroup, including the control group, were anesthetized, euthanized, and dissected. Different body tissues, and the kidney, liver, bones, and heart, were isolated and preserved in formalin solution for subsequent analysis. CBC, liver, renal, and cardiac biomarkers were evaluated. In addition, histopathology and bone characteristics of soft tissues were also conducted. Mortality and morbidity were recorded. The result showed significant disruptions in CBC and other biomarkers related to kidney, liver, and heart in a dose-dependent pattern. Although vitamin supplementation improved the overall outcome, the improvement was not statistically significant. Histopathological examination displayed changes in the exposed (E) group, with no observable improvements with vitamin supplementation. Moreover, the bone-related parameters exhibited similar trends.