The oil price instability and the measures taken to reduce the increase in greenhouse gas emissions are the main factors promoting the development and use of environmentally friendly energies. From an energy efficiency point of view, biofuels constitute a renewable energy source and its use helps to reduce energy dependency on fossil fuels. The most used biofuels for transport worldwide are biodiesel (BD) and bioethanol. However, there are other options such as straight vegetable oil (SVO). SVO can be small-scale produced in local cooperatives through pressing, filtering and conditioning processes which are much simpler than the ones required for BD production. In this study a comparative life cycle assessment (LCA) of two biofuels obtained from Spanish rapeseed, namely small-scale SVO and large-scale BD, is performed. The LCA methodology allows the two biofuels' production and their rate of consumption in a vehicle (a truck) to be compared. In this manner, it is possible to assess which is environmentally advantageous: to use SVO directly as biofuel or to convert it to BD. Moreover, LCA is used in the study to calculate the energy return on investment index (EROI) and an energy conversion ratio to evaluate which biofuel is more energy efficient. The obtained results show the environmental benefits of using SVO instead of BD by analyzing representative impact categories defined by the CML and EDIP methods. A sensitivity analysis has also been conducted. EROI indexes for SVO and BD production show a clear preference for SVO as compared to BD. (C) 2010 Elsevier Ltd. All rights reserved.