This research aims to evaluate the influence of pyrolysis time and temperature on the composition and properties of bio-oil derived from Mimusops elengi. Experiments were conducted by varying the pyrolysis temperature and time from 400 to 600 °C and 30 to 120 min, respectively. Both pyrolysis temperature and time were found to significantly influence the bio-oil composition. At enhanced pyrolysis temperatures, the bio-oil yield increased while the ash and gas yields decreased. In addition, extended pyrolysis time produced a greater bio-oil yield, indicating that higher temperatures and longer durations promote additional decomposition of biomass. Functional groupings, including alcohols, phenols, ketones, esters, and aromatic compounds in the bio-oil, were identified via FT-IR analysis, indicating that the bio-oil’s diversified chemical properties make it a potential alternative feedstock. GC-MS analysis identified 26 chemical compounds in the bio-oil, of which phenol was the most abundant. However, a high phenol content can diminish bio-oil quality by enhancing acidity, decreasing heating value, and encouraging engine corrosion. Temperature and pyrolysis time are crucial factors in producing bio-oil with the desired chemical composition and physical properties. The maximum yield, 34.13%, was attained after 90 min of operation at 500 °C. The characteristics of the Mimusops elengi bio-oil produced, namely density, viscosity, pH, and HHV were 1.15 g/cm3, 1.60 cSt, 4.41, and 19.91 MJ/kg, respectively, in accordance with ASTM D7544. Using Mimusops elengi as a pyrolysis feedstock demonstrates its potential as an environmentally friendly energy source for a variety of industrial and environmental applications. The yield of bio-oil produced is not optimal due to the formation of tar, which results in the blockage of the output flow during the pyrolysis process.