Nanofertilizers offer a promising solution to increase crop yields amid increasing population pressure. Yet  there have been safety concerns about their use, particularly in challenging conditions. This work was designed to assess the toxicological effects of a chelated multi-micronutrient nanofertilizer on albino rats (Rattus norvegicus) as non-target organisms, focusing on the liver, which has important metabolic functions. Thirty-five animals were weighed and randomly divided into 5 groups (n=7). A negative control group was included, and four experimental groups (C1-C4) were given the test item. The dosing regimen for the experimental groups was 15 oral doses of nanofertilizer every other day for 29 days. All the groups were treated with different doses (62.5, 125, 250, and 500 mg/kg, respectively). Biochemical analyses revealed a statistically significant reduction in lactate dehydrogenase (LDH) levels in groups C2 through C4 (661.00 ± 181.67, 572.43 ± 187.62, and 577.86 ± 110.84 U/L, respectively) compared to both the control (919.86 ± 232.08 U/L) and C1 (880.00 ± 119.17 U/L) groups. In contrast, serum total protein concentrations exhibited a marked increase across all treatment groups (C1–C4: 74.36 ± 3.91, 68.49 ± 5.88, 70.47 ± 7.93, and 68.14 ± 3.84 g/L, respectively), relative to the control (59.29 ± 5.49 g/L). This rise in total protein was mainly due to a significant increase in serum globulin, whereas albumin showed a marked decrease. Histological examination showed dose-dependent hepatotoxicity in all treatment groups. These results indicated that a sub-lethal dose of this nanofertilizer has the potential to elicit observable hepatotoxicity, underscoring the need for careful application and further toxicological evaluations.