The purpose of this study is to investigate the effects of fuel temperature and nozzle length‐diameter ratio (L/D) on biodiesel spray characteristics, under high injection pressure and high ambient pressure, by numerical method. The analysis of spray characteristics is carried out in conjunction with the transient flow inside nozzle. To achieve this, three‐dimensional calculation grids of spray nozzle and spray domain were setup, and the needle movement was achieved by dynamic mesh technique. The reliability of spray models was validated by experimental results. It was established that the variation in spray tip penetration (STP) and Sauter mean diameter (SMD) is similar at different initial conditions. The STP increases at a faster rate initially and then slows down, whereas the SMD gradually decreases with time after the injection. In addition, a sensitivity analysis of the effects of injection parameters on biodiesel spray characteristics was conducted. Compared to the nozzle L/D, injection pressure and fuel temperature have a greater impact on biodiesel spray characteristics. The increase in injection pressure has a significant effect on the velocity distribution, STP and SMD. When the injection pressure is increased from 100 to 200 MPa, the maximum velocity of the spray core zone increases by 33.96%, the STP increases by 27.17%, and the SMD reduces by 14.81%. Furthermore, an increase in fuel temperature mainly affects the concentration distribution and the SMD of atomized droplets. When the fuel temperature is increased from 300 to 350 K, the maximum concentration of axial spray center lowers by 25.41%, and the SMD decreases by 17.19%. However, the increase of L/D chiefly impacts the concentration of axial spray center, but has little effect on other spray parameters. When the nozzle L/D is increased from 4 to 8, the maximum concentration of axial spray center increases by 20.29%.