Background and objective Changes in dietary habits are associated with an improvement of lipid profile. We decided to assess changes in lipid profile reached with a dietary enrichment with canned tuna in olive oil and compared them with those reached with canned tuna with ad-ditional supplementation. Subjects and method In a prospective, randomized, single blind study, 400 participants were assigned to a daily intake of canned tuna in olive oil alone (control group) or enriched with: isoflavones, omega-3 fatty acids of phytosterols, during three months. Plasmatic lipid levels were measured before and after intervention. Changes of lipid levels between basal and final visits and differences between the 4 types of supplementations were evaluated. Results The mean age of the study population was 53 years, 45% of them being males. In all 4 groups a significant reduction of total cholesterol, low-density lipoprotein (LDL)-cholesterol levels and total cholesterol/high-density lipoprotein (HDL)-cholesterol ratio was observed after 3 months of follow-up; there were no significant changes of triglycerides nor HDL-cholesterol levels. The comparison between different types of supplementation revealed that only phytosterols addition reached a greater reduction of total cholesterol and LDL-cholesterol levels than canned tuna in olive oil alone (p < 0.05), and the differences between the remaining 2 supplementations and the control group were not significant. Conclusions Regular intake of enrichment with canned tuna supplementation is associated with improvement of lipid profile. The addition of isoflavones, omega-3 fatty acids or phytosterols supplementations increases fish consume effect, although only the enrichment with phytosterols reaches significantly better results.