Background: Argania spinosa seeds oil is an important endemic product of Morocco that is used traditionally as nephroprotective agent. Aim: Our study aims to compare the chemical compositions of unroasted (UnRoil) and roasted (Roil) Argania spinosa seed oils and their nephroprotective activity against gentamicin-induced kidney damage in Wistar rats. Experimental procedure: The chemical contents of argan oil samples were analyzed by GC/MS analysis and the nephroprotective activity was assessed by administering gentamicin to Wistar rats. Biochemical and histopathological examinations were performed on the kidneys and the blood samples. Results: The results illustrate the presence of four shared fatty acids, namely palmitic, stearic, linoleic, and oleic acids in both Roil and UnRoil, with comparable amounts. Additionally, the findings revealed that Roil contains a specific compound, heptadecane, 2,6-dimethyl-. Both the animals in Roil + Gentamicin and UnRoil + Gentamicin groups exhibited significant reductions in water intake, urinary volumes, food intake, and serum levels of creatinine, urea, urine sodium, and MDA compared to the gentamicin-treated group. However, urine urea, urinary creatinine, catalase, and glutathione levels exhibited significant increases. Moreover, the oils maintained normal left kidney weights compared to the gentamicin-treated group. Histopathological analysis supported these results, indicating that administering 2 ml/kg/day of UnRoil or Roil prevented significantly gentamicin-induced alterations in the glomerulus. Conclusion: Our findings confirm favorably the use of Argania spinosa in traditional Moroccan medicine as a nephroprotective agent. Interestingly, we observed that the roasting process of its seeds did not affect this pharmacological effect. This suggests a novel approach to harness the beneficial effects of a phytomedicine tailored for renal issues.