Physalis peruviana L. is widely used in folk medicines to manage diabetes. This study aimed to identify new chemical constituents and explore in vitro antioxidant ability and alpha-glucosidase inhibition activity to explain its antidiabetic mechanism. Methanolic extracts and hexane fractions were prepared and analyzed by CG-FID and GC-MS. In vitro antidiabetic activity was assessed on α-glucosidase inhibition and DPPH models. The phytochemical analysis detected 29 constituents, manly heptacosane (25) and hexadecanoic acid methyl ester (14). Aliphatic hydrocarbons and esters were identified as the main classes of secondary metabolites. Compared to the standard, the methanol-water fraction was the most potent fraction on α-glucosidase inhibition (IC50 32.5 ± 0.27 µg/mL). The antioxidant values (IC50) of two fractions, and the reference were quite close (44.5 ± 0.54; 49.5 ± 0.87; 44.2 ± 0.77 µg/mL). Almost 82.75% are recognized for the first time in the leaf hexane fractions of this plant. The observed antioxidant activity was found to be significantly notable. Manuscript profile