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Biodiesel made from Jatropha oil by transesterification process has viscosity and other important physical properties comparable to mineral diesel hence it can be used as an alternate fuel in conventional diesel engines. It is important to investigate the spray characteristics of biodiesel because emissions from the engines are dependent on fuel atomization process and resulting fuel-air mixing. This study focuses on the Jatropha biodiesel spray investigations using Phase Doppler Interferometry (PDI) for measurement of various microscopic spray parameters such as Sauter mean diameter (SMD) and spray droplet size and velocity distributions. The spray and engine experiments were carried out for Jatropha biodiesel (JB100) and their 20% blends (JB20) with mineral diesel as baseline. Fuel injection pressure during the spray experiments was maintained at 200 bars for all tests, quite similar to small horse power agricultural engines, and the fuel injection quantity was varied.