<p>Biofuel, including biodiesel derived from animal adipose tissue, emerges as a renewable energy alternative, markedly mitigating airborne toxic pollutants by exceeding a 50% reduction. In the present research, animal fats, possessing heightened efficacy in biodiesel synthesis, undergo transesterification to refine fuel characteristics, rendering them apt for biofuel applications. Samples of cattle fat from mesentery, pericardium, and abdominal sectors underwent comprehensive analysis encompassing assessment of oil yield, acid number, free fatty acid content, and biodiesel manufacturability. The obtained oils were recorded as 39.46%, 38.74%, and 38.71% from the mesentery, pericardium, and abdominal segments. Gas Chromatography-Flame Ionization Detector (GC-FID) discerns the fatty acid composition, revealing conspicuous concentrations of Palmitic Acid(C_16:0), Stearic Acid(C_18:0), and Linoleic Acid(C_18:1), indicative of major fatty acid constituents. Transesterification of rendered fat ensured biodiesel synthesis, employing alkaline, acidic, and lipase catalysts. Analyses of the data using the single-factor ANOVA test calculated the mean values ± standard deviation for variables. The greater efficacy of alkaline catalyst produced biodiesel roughly 38.56%, 38.95%, and 39.28%, from mesentery, pericardium, and abdominal fat respectively. Gas Chromatography-Mass Spectrometry (GC-MS) examination was performed on a biodiesel sample obtained through an alkaline catalyst. Based on retention duration, each component was identified, exposing the nature of fatty acid methyl esters. This investigation revealed the value of animal fat as a substitute source for the manufacturing of biodiesel. This kind of feedstock was found as easily affordable, cost-effective, energy-rich, and readily available from an economic and commercial perspective. It was assessed as a renewable source in the sense that it is easily replenishable along with least environmental impacts.</p>