Diabetes occurs due to increased glucose levels in the blood. The glycemic rises over a long period; it will damage many organs, especially the heart, kidneys, and eyes. Inhibiting α-amylase and α-glucosidase activity will slow down the process of breaking down starch into glucose, helping to prevent hyperglycemia after eating. Annona glabra L. (Pond Apple) is a common species in Vietnam in the Annona genus. This genus has compounds of alkaloids, flavonoids, tannins, and saponins, and some species in it have anti-diabetic activity. It has yet to be studied in Annona glabra L. The extracts of 96% ethanol (96 ET), 50% ethanol (50 ET), and aqueous (AE) are extracted from the bark stem of Pond Apple; semi-qualitative of alkaloids and flavonoids; investigation inhibitory activity of α-amylase and α-glucosidase in vitro. Extract of 50% ethanol has higher alkaloids and flavonoids than the others. The α-amylase and α-glucosidase inhibitory activity of 50 ET were 0.05 mg/mL and 0.09 mg/mL; 96 ET, respectively, 0.07 mg/mL and 0.05 mg/mL. This result opens the potential for diabetes treatment products that are naturally derived, easy to find, low cost, and have fewer side effects.