Anastrepha fraterculus (Wiedemann), Ceratitis capitata (Wiedemann), and Drosophila suzukii (Matsumura) are the main fruit pests in Brazil. Here, we evaluated the susceptibility of Brazilian populations of A. fraterculus, C. capitata, and D. suzukii to selected insecticides. In ingestion bioassays, adults from a laboratory susceptible population of each species were exposed to five different modes of action of insecticide. Then, field populations of each species were exposed to the diagnostic concentrations to evaluate possible changes in susceptibility. Our findings indicate that lambda-cyhalothrin, malathion, and spinosad had similar levels of toxicity against a susceptible laboratory population of A. fraterculus, with LC₅₀ values of 6.34, 6.54, and 8.76 µg a.i./ml, respectively. Ceratitis capitata had similar susceptibilities to spinosad (1.30 µg a.i./ml), spinetoram (2.76 µg a.i./ml), and malathion (7.10 µg a.i./ml), but a lower susceptibility to lambda-cyhalothrin (76.55 µg a.i./ml). For D. suzukii, the LC₅₀ values of deltamethrin (0.67 µg a.i./ml), malathion (3.30 µg a.i./mL), spinosad (4.16 µg a.i./ml), and spinetoram (4.75 µg a.i./ml) were lower than for abamectin (15.02 µg a.i./ml), acetamiprid (39.38 a.i./ml), and thiamethoxam (70.15 µg a.i./ml). The diagnostic concentrations, based on LC₉₉ values of the insecticides, caused more than 99% mortality for most field populations of each species. For lambda-cyhalothrin the populations RS-1 (A. fraterculus) and SC (C. capitata) showed approximately 10% of live insects. These differences represent the natural variation in population susceptibility and not due to insecticide selection pressure. The diagnostic concentrations defined here should be used in future resistance monitoring programs in Brazil.