Transport consumes around one-third of Latvia's final energy demand being the largest sector that produces greenhouse gas emissions. In Latvia, in the year 2019, the share of renewable energy sources in the transport sector was only 4.8 % of the total gross consumption with a target of 7 % of not food-based biofuels by 2030. To tackle climate change problems, one significant task is reducing pollution in the transport sector by substituting fossil with biomass-based fuels. The goal of this study is to develop a methodology for evaluating biofuel production in Latvia considering economic, social, environmental, and technological aspects. A total of 16 specific criteria were selected for biofuel comparison. The methodology is based on a combination of two quantitative approaches, namely Life Cycle Assessment and Multi-Criteria Analysis. The proposed method aims to identify the most sustainable biofuel in Latvia according to a set of specific indicators.