Physicochemical parameters of pasta enriched with tilapia (Oreochromis niloticus) flour were investigated. Five formulations were prepared with different concentrations of tilapia flour as partial substitute of wheat flour: pasta without tilapia flour (PTF0%), pasta with 6% (PTF6%), 12% (PTF12%), 17% (PTF17%), and 23% (PTF23%) of tilapia flour. The formulations were assessed for proximate composition, fatty acid and amino acid profile on day 1 whereas, instrumental color parameters (L*, a* and b* values), pH, water activity (aw), and lipid and protein oxidation were evaluated on days 1, 7, 14, and 21 of storage at 25°C. Fortification with tilapia flour increased (p < 0.05) protein, lipid, ash, total essential amino acids, and total polyunsaturated fatty acids contents. In addition, supplementation of pasta with tilapia flour decreased (p < 0.05) lightness and water activity while redness, yellowness, pH values, and lipid oxidation were increased (p < 0.05) in a level-dependent manner. Nevertheless, all formulations were exhibited storage stability at 25°C. In general, protein oxidation was greater (p < 0.05) in the pasta containing 12%, 17%, and 23% of tilapia flour than their counterparts, and the storage promoted an increase (p < 0.05) on the carbonyl content in all formulations. Thus, pasta with 6% of tilapia flour has the potential to be a technological alternative to food industry for the nutritional enrichment of traditional pasta with negligible negative effects on the chemical stability of the final product during 21 days at 25°C.