The effects of seasonal and regional variation on the proximate and fatty acid compositions of sardine (Sardina pilchardus) were determined. The levels of lipid displayed pronounced seasonal than regional fluctuations with the highest values in summer and the lowest values in winter for both areas (Kelibia and Teboulba, Tunisia). Negative correlations between fat and water contents were observed. The higher water contents were recorded during winter for sardine of Kelibia and Teboulba (78.5 +- 0.86% and 77 +- 0.5%, respectively). For both areas, the samples were not influenced by seasonality in terms of their total fatty acid (p>0.05). The main fatty acids of the total lipid were C16: 0, C14: 0, C18: 0, C18: 1, DHA C22: 6n3, EPA C20: 5n3 and DPA C22: 5n3. Sardina pilchard us muscle contained appreciable levels of omega-3 polyunsaturated fatty acids suggesting the use of this fish as a source of healthy diet for humans.