When oils or fats sourced from plants or animals undergo transesterification with short-chain alcohols like methanol or ethanol, the resulting product is biodiesel—an alkyl ester of fatty acids. International organizations, including ASTM (American Society for Testing and Materials), CEN (European Committee for Standardization), and BIS (Bureau of Indian Standards), have adopted and regularly updated biodiesel specifications and recommendations. Vegetable oils of edible grade, such as soybean, rapeseed, and sesame, can be partially or entirely processed and utilized as feedstock for biodiesel production. Sesame oil, subjected to chemical extraction and employing homogeneous basic catalysis for transesterification, yields biodiesel with physical and chemical characteristics akin to mineral diesel, meeting ASTM and English standards. In comparison to biodiesel derived from other vegetable oils, transesterification-derived biodiesel from sesame oil exhibits superior oxidation stability, attributed to the presence of special compounds called lignans. These naturally occurring compounds in sesame oil, including sesamine, sesamoline, and sesamol, contribute to its enhanced oxidation stability. Analyzing sesame oil and biodiesel involves various analytical methods, such as chromatography, spectroscopy, nuclear magnetic resonance, and thermogravimetric behavior. The transesterification parameters, such as catalyst type and temperature, significantly impact biodiesel yield. The study supports the viability of using petrodiesel as a substitute for methyl ester derived from sesame plants, backed by quantitative and qualitative findings. According to this research, sesame plants emerge as a valuable biomass energy source due to their ease of cultivation and high oil yield.