Five species of fruit flies of the genus Anastrepha are recorded for the first time from Colombia: A. alveata Stone, 1942, A. antunesi Lima, 1938, A. palae Stone, 1942, A. parishi Stone, 1942, and A. shannoni Stone, 1942. The species were collected from the municipalities of Arauca (department Arauca), Cerete (department Cordoba), and La Mesa (department Cundinamarca). Information is also provided about the distribution and morphological variation of these species.