This study investigated the effects of replacing beef tallow with canola oil (CA) on the quality characteristics and storage stability of pemmican. Six treatment groups were established: CON (beef tallow), CA1 (2.5% CA), CA2 (5.0% CA), CA3 (7.5% CA), CA4 (10% CA), and CA5 (12.5% CA). Samples were stored at 4°C for 2 weeks. Fatty acid composition, proximate composition, color, water activity (aw), and texture profile analysis (TPA) were assessed. Additionally, pH, volatile basic nitrogen (VBN) and 2-thiobarbituric acid reactive substances (TBARS) were measured at 0, 7, and 14 days. The levels of saturated fatty acids decreased with higher CA content, while oleic acid and linoleic acid increased. Results showed that aw decreased with increasing CA levels (p<0.05). TPA indicated decreasing trends in hardness, chewiness and gumminess as CA levels increased. During the first week of storage, pH decreased for CON and CA1 but increased for CA2-5 (p<0.05). CA5 exhibited the highest TBARS throughout storage period, suggesting increased lipid oxidation. VBN showed a decreasing trend with higher CA levels, indicating improved protein stability. Therefore, replacing beef tallow with CA can improve the composition of fatty acid and storage characteristics. Based on the findings, CA4 emerged as the most suitable option. These findings provide valuable insights into optimizing pemmican formulation for improved quality and shelf life.