The scarcity of the fossil fuel, environmental pollution and food crisis are the world's major issues in current era. Biodiesel is an alternative to diesel fuel, environment friendly and biodegradable and is produced from either edible or non-edible oils. In this study, a non-edible rubber seed oil (RSO) with high free fatty acid (FFA) content of 45 were used for the production of biodiesel. The process comprises of two steps. The first step is the acid esterification to reduce the FFA value and the second step is the base transesterification. The response surface methodology (RSM) was used for parametric optimization of the two stage processes i.e. acid esterification and base transesterification. The yield of biodiesel was analyzed using gas chromatography. The FTIR (Fourier Transform Infra-Red) spectrum was also determined to confirm the conversion of fatty acid to methyl esters. The fuel properties were analyzed according to the ASTM D6751 and EN14214 and were compared with the previous finding of researchers. All analyzed properties fulfilled the biodiesel standard criteria. © 2013 Elsevier Ltd. All rights reserved.