Callus cultures of Annona muricata and Annona purpurea were induced in Murashige and Skoog (MS) medium supplemented with different concentrations of 1-naphthylacetic acid (NAA), 6-benzyladenine (BA) and 2,4dichlorophenoxyacetic acid (2,4-D) utilized hypocotyls with explant. The highest percentage of callus formation was the treatment supplemented with 3 mg L-1 NAA for A. muricata (100%) while for A. purpurea in lower percentage (75%). BA stimulated the formation of shoots in all the evaluated concentrations, being the concentration of 2 mg L-1 the one that induced the greater formation of shoots for A. muricata (23 shoots/explant) and A. purpurea (28 shoots/explant). The content of total phenols, flavonoids and antioxidant activity was measured in the callus obtained from both species. The results showed that a higher content of total phenols was quantified in callus of A. purpurea (27.8 mg g-1 dw) compared to A. muricata (23.2 mg g-1 dw). The highest content of total flavonoids was observed in the callus of A. purpurea (8.0 μg g-1 dw). Antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydracil radical assay. The concentration required for 50% inhibition (IC50) of the 2,2-diphenyl-1-picrylhydracil radicals were 4.22 μg mL-1 in methanolic extracts of callus of A. muricata, while in extracts of callus of A. purpurea was 2.86 μg mL-1 , in both cases was greater than that found for leaves. Callus culture of the species studied in this work represents an alternative for the production of natural antioxidants.