The aim of this work is to study the influence of the physicochemical characteristics of neem seeds, according to their mass and oil content, on the production of biodiesel. After the physical characterization of the seeds and extraction of the oil (triglycerides), biodiesel was produced from crude neem seed oil by transesterification with ethanol in the presence of sodium hydroxide. This study shows that the physicochemical characteristics of these seeds vary according to the origin of the samples. The seeds from Zidim, with a mass average of 200 seeds evaluated at 141.36 g and an almond content of 40.70%, have better characteristics compared to those collected in the city of Maroua, with average values evaluated at 128.00 g and 36.05%, respectively. Almonds have an average lipid content of 53.98 and 56.75% for the Maroua and Zidim samples, respectively. This study also reveals that neem oil, by its physicochemical characteristics, has a satisfactory quality for a valorization in the production of biodiesel. However, its relatively high free fatty acid content is a major drawback, which leads to a low yield of biodiesel, evaluated on average at 89.02%, and requires a desacidification operation to improve this yield. The analysis of biodiesel indicates physicochemical characteristics close and comparable to those of petrodiesel, particularly in terms of calorific value, density, kinematic viscosity, acid value, evaluated at 41.00 MJ/kg, 0.803, 4.42 cSt, and 0.130 mg/g, respectively.