This work gives a comprehensive chromatographic assessment of biodiesel generation from plant seed oil using ecologically friendly nano‐catalysts. Researchers all over the world are actively looking for new ways to satisfy the urgent need for clean and renewable energy sources. The resultant biodiesel was fully characterized utilizing modern techniques like scanning electron microscopy, energy diffraction X‐ray and X‐ray diffraction. The biodiesel gas chromatography/mass spectrometry analysis revealed four significant peaks of fatty acid methyl esters, indicating high‐quality biodiesel production. Furthermore, the biodiesel fuel qualities were discovered to be comparable with international standards such as ASTM D‐6571 and EN‐14214. This indicates that the iron‐modified clay nano‐catalyst can be used as a catalyst for large‐scale biodiesel production. This work is important because it could lead to the large‐scale production of a novel, non‐food feedstock. We may lessen our reliance on fossil fuels and contribute to a more sustainable and ecologically friendly energy future by leveraging the usage of biodiesel produced in this way. The chromatographic assessment of biodiesel production from non‐edible seed oil using environmentally benign nano‐catalysts holds significant promise in advancing sustainable and eco‐friendly biodiesel production methods, contributing to a cleaner and more environmentally responsible energy sector.