This work addressed the traditional nixtamalization processing conditions on the in vitro protein and starch digestibility of tortillas. Two levels of lime addition (1 and 2 g/100 g db) and three different cooking times (30, 45 and 60 min) were considered. FTIR analysis was used to evaluate the organization of water, proteins and starch in the tortilla matrix. Lime concentration impacted the random secondary structure formation of the protein, and cooking time the starch ordered structures content. Higher lime concentration led to lower protein digestibility, but longer cooking time increased it. Highest protein hydrolysis (74.36 ± 3.72%) was achieved in T