This study was conducted to investigate the influence of smoking time (3 and 6 days) on the shelf life of cold-smoked kutum stored for 60 days at room temperature (25 ± 2°C) via biochemical quality indices and fatty acids content analyses. The results obtained from peroxide value and thiobarbituric acid-index analyses indicated that prolongation of smoking time and preservation duration resulted in a higher degree of lipid oxidation (P < 0.05). Comparison between the treated samples mentioned less reduction (P > 0.05) in the pH values of 3 days-smoked kutum during the storage for 60 days. However, based on the results of total volatile base nitrogen, microbial spoilage significantly increased (P < 0.05) and exceeded above the acceptability limit when smoking time decreased to 3 days. The kutum smoked for 6 days were contained higher percentage (P > 0.05) of polyunsaturated fatty acids during storage, whereas those smoked for 3 days mentioned higher percentages of saturated fatty acids and monounsaturated fatty acids. Polyunsaturated fatty acids/saturated fatty acids ratio of both smoked groups was within the recommended values for the human diet. This study concluded that prolongation of the smoking time led to increase the shelf life of cold-smoked kutum with higher nutritional value during 60 days storage at room temperature.