This study aimed at elucidating the influence of food matrix on the release of antioxidant activity from five plant foods (apple, spinach, walnut, red bean, and whole wheat). To this purpose a protocol based on sequential enzymatic digestion was adopted. The total antioxidant capacity (TAC) of both solubilized and insoluble materials was measured at each step. Results showed that the overall TAC obtained by enzyme treatments was usually higher than that obtained by chemical extraction-based methods. In apple most of the TAC was released upon water washing and after pepsin treatment, whereas in spinach, beans, and whole wheat the TAC released by treatments with bacterial enzymes was prominent. Walnut had the highest TAC value, which was mainly released after pancreatin treatment. Therefore, the enzyme treatment is fundamental to estimate the overall potential TAC of foods having a high amount of polyphenols bound to dietary fiber or entrapped in the food matrix.