The use of nanomaterials in agriculture is a current need and could be helpful in overcoming food security risks. Brassica napus L. is the third most important crop for edible oil, having double low unsaturated fatty acids. In the present study, we investigated the effects of green synthesized Zn NPs on biochemical effects, antioxidant enzymes, nutritional quality parameters and on the fatty acid profile of rapeseed (B. napus). Plant-mediated synthesis of zinc nanoparticles (Zn NPs) was carried out using Mentha arvensis L. leaf extract followed by characterization through ultraviolet–visible spectroscopy (UV-vis), scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy dispersive X-Ray (EDX), and X-Ray diffraction (XRD). NPs exhibited irregular shapes ranging in size from 30–70 nm and EDX analysis confirmed 96.08% of Zn in the sample. The investigated biochemical characterization (protein content, proline content, total soluble sugar (TSS), total flavonoid content (TFC), and total phenolic content (TPC) showed a substantial change on exposure to Zn NPs. A dose-dependent gradual increase was observed in the antioxidant enzymes, superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). Oil and moisture contents dropped significantly from the control level in the rapeseed (B. napus) varieties. However, different trends in nutritional (Zn, Na+, K+) and fatty acid profiling of B. napus have been noted. This study demonstrates that Zn NPs have the potential to improve the biochemical, nutritional, antioxidant enzymes, and fatty acid profile of B. napus varieties.