Canola biodiesel (B100) may have improved cold flow properties when compared to other biodiesel feedstocks which is attributed to canola’s high unsaturated and low saturated fat content. Production environment and plant genetics influence fatty acid composition, but the influence of environment and canola variety selection on B100 quality is not clearly understood. The objective of this study was to evaluate canola fatty acid composition and biodiesel quality among several ND locations. In Exp 1, bulked canola samples from two ND locations across five growing seasons (2003-2007) were analyzed and Exp 2 evaluated a single canola variety (Interstate Hyola 357) harvested at two locations (2003, 2004, 2005, and 2007). In both experiments, B100 was produced directly from seed via in situ alkaline transesterification methods. The impact of production year and location on canola fatty acid composition and several B100 ASTM D6751 quality standards including cloud point and oxidative stability index is presented.