The search for sensitive and rapid analytical techniques for the determination of natural antioxidants is an area in constant growth due, among other aspects, to the complexity of plant matrices. In this study, silver nanoparticles prepared with the aqueous extract of Mimosa albida leaves were used to assess their polyphenolic content and antioxidant capacity. Silver nanoparticles were characterized by different techniques. As a result, nanoparticles of 6.5 ± 3.1 nm were obtained. The total phenolic content in the extract was 1320.4 ± 17.6 mg of gallic acid equivalents GAE· 100 g−1 and in the nanoparticles 257.3 ± 5.1 mg GAE· 100 g−1. From the phenolic profile analyzed by ultra high-performance liquid chromatography (UPLC) with a diode-array detector (DAD), the presence of apigenin and luteolin in the plant extract is postulated. The antioxidant capacity measured by oxygen radical absorbance capacity ORAC-fluorescein assay was 86917 ± 6287 and 7563 ± 967 µmol ET g−1 in the extract and nanoparticles respectively. Electrochemical analysis by cyclic voltammetry (CV) confirmed the effective reduction capacity of the Mimosa albida leaves extract to reduce Ag ions to AgNPs and differential pulse voltammetry (DPV) suggested the presence of two main reducing agents in the extract. From this study, it was concluded that the aqueous extract of Mimosa albida contains reducing agents capable of synthesizing silver nanoparticles, which can be used in the phytochemical industry.