Biodiesel mainly comes from edible oil, and there is little research on its yield from non-edible sources with low-cost oil. It is paramount to investigate the non-edible oil resources which may lead to advance the commercial feasibility of biodiesel and cost effectiveness as well as resolve the food issues. This chapter describes four novel non-edible seed oil sources comprising Koelreuteria paniculata, Rhus typhina, Acacia farnesiana and Albizzia julibrissin for biodiesel production. We aimed to optimize different reaction parameters for oil extraction, alkali-catalyzed transesterification process for maximal biodiesel production and finally evaluate its compatibility with mineral diesel. The optimization factors in transesterification included the molar ratio of methanol to oil, reaction time, stirring intensity, catalyst concentration and temperature. Two methods have been described including Soxhlet and mechanical for extraction of seed oil. The synthesized esters were evaluated and characterized through the nuclear magnetic resonance (NMR; 1H and 13C), Fourier transform infrared (FT-IR) and gas chromatography–mass spectrometry (GC–MS) and the total conversion of crude oil to fatty acid methyl esters (FAMEs) were established. The inductively coupled plasma-optical emission spectrometry (ICP-OES) and Elemental Analyzer (EA) were used for evaluation of elemental concentration. The physico-chemical characterizations of the biodiesel, i.e., flash point, pour point, cloud point, and density were within the American Society for Testing and Materials (ASTM; D6751) and European Standards ((EN14214). Koelreuteria paniculata produced highest biodiesel oil content by Soxhlet extraction (28–30%) followed by the Albizzia julibrissin (19–24%), Acacia farnesiana (23%), Rhus typhina (20–22%). The density ranged from 0.83–0.87 @ 15°C (g/cm3) and the kinematic viscosity ranged from 3.75–6.3 (mm2/s) among all the plant sources. Koelreuteria paniculata had highest Na (5456.2), Cr (1246.8), Ni (658.36), and Al (346.87) elemental concentrations (μg/g) than other plant sources. The elemental percent of C, H, N, and O of biodiesel ranged from 72.54–76.86, 11.25–13.34, 1.97–2.73, and 9.86–12, respectively. In conclusion, these non-edible plant seeds offer a cheap source of renewable energy and can be easily grown on barren and wastelands and contribute to efficient biodiesel production to mitigate the energy crisis.