Biodiesel from plants can compete economically with petroleum diesel fuels as a future prospective fuel. The bio-oil was extracted from the plant of Aloe barbadensis by soxhlet extractor using hexane as an organic solvent, and then further, the oil was analyzed for moisture content, pH, specific gravity, density, viscosity, saponification value, refractive index, peroxide value, acid number, free fatty acid, and iodine value. The results showed that four major fatty acids, like nonadecanoic acid, n-hexadecanoic acid, octadecadienoic acid and octadecenoic acid were present in the bio-oil of Aloe barbadensis and characterized by GC-MS, which showed that the bio-oil could be used for producing high quality of biodiesel. In furture studies, converted crude biooil into biodiesel using hetergenous catalyst will be anlaysed for their engine characteristic studies.