The role of liquid biofuels in transportation to minimize the effects of climate change is  evident and has led to a number of studies on finding effective solutions to replace fossil fuels. Liquid biofuels are especially important for heavy duty transports as the effective ‘green’ alternatives are not as many compared to light duty vehicles; for which for e.g. electrification is an option. This thesis presents a comparison study of 8 liquid biofuels with a total of 13 different fuel pathways for use in road freight transports; both current and potential future fuels are assessed in terms of their environmental effects, fuel properties and compatibility with the heavy duty vehicle engines (see Table 10, page 36). Furthermore, a case study is performed to assess the practicality of the results of the study.  Hydro-treated vegetable oil, Bio Dimethyl ether, Liquefied Bio Methane/ ED95 are identified as fuels with considerable potential in the shorter term. Algal biofuel and Biomass to liquid (BTL) fuels from synthesis gas, if realized commercially would be a breakthrough for biofuels in overall transportation sector. However, life cycle analysis has to be performed for the different fuel pathways to completely understand the various impacting factors.