The reduction of saturated fats in canola oil has recently been promoted as a goal for breeders for commercial and human health benefits. Currently, saturated fatty acids in Canadian produced canola oil are above the 7% level, and the objective of this study was to generate canola lines with reduced major saturates (palmitic and stearic), by several percent. Mutant embryos generated from direct ultraviolet radiation mutagenesis of microspores in vitro were subjected to heat during the maturation stage. Heat artificially elevated the saturate levels in developing mutant embryos, allowing efficient identification of those with reduced saturates within the expanded range using HPLC fatty acid analysis of the embryo cotyledons. Mutagenesis produced embryos with fatty acids altered in both directions. Major saturate levels in the cotyledons of heat-treated mutant embryos ranged from 3.3 to 16.4% (heated control ca. 6-9%) and 1.3-10% (heated control ca. 2-4%) for palmitic and stearic fatty acids, respectively. Doubled haploid seed derived from embryos grown at normal temperatures confirmed the reduction of major saturates. HPLC fatty acid analysis of DH seed identified saturate levels ranging from 3.9 to 6.5% (control ca. 5.5%) and 0.9-2.7% (control ca.