Concerns have been raised about the international impacts of biofuel projects, including higher food prices, as well as indirect land use change which contribute to a negative carbon footprint.  This has led discussions around the sustainability criteria required for renewable energy policy biofuel policy.  As the proportion of renewable energy within total primary energy supply increases, these criteria will become more important.  A regional analysis methodology is used to explore the likely impacts of renewable fuel development.  Eastern Ontario is selected as a region that has significant potential for biomass production, including both agricultural and forest feedstocks.  The potential for biomass production across all lands within this region have been mapped and quantified, as have the human capital resources found within the local population.  Potential platforms for biofuel generation, including 1^st- and 2^nd-generation technologies, were used to build potential scenarios of future biofuel use.  The impacts of existing and potential biofuel development are measured using environmental, economic, and social parameters.  Specific tools applied include life cycle assessment and techno-economic modeling, which quantify economic and environmental measures to inform policy assessment and recommendations.  This paper also considers recent international developments in measuring biofuel sustainability, including efforts at the European Commission level as well as examples from individual countries.  Models for ensuring sustainability, including certification systems, are discussed.  In order to inform a framework that will permit the bioenergy industry to develop at a rapid but sustainable rate, criteria for measuring the sustainability of biofuel solutions at the regional level are proposed.