Cerumen is a bee product produced exclusively by stingless bees, resulting from a mixture of beeswax and plant resins. The antioxidant activity of bee products has been investigated since oxidative stress is associated with the onset and progression of several diseases that can lead to death. In this context, this study aimed to investigate the chemical composition and antioxidant activity of cerumen produced by theGeotrigonasp. andTetragonisca fiebrigistingless bees, in vitro and in vivo. The chemical characterization of cerumen extracts was performed by HPLC, GC, and ICP OES analyses. The in vitro antioxidant potential was evaluated by DPPH•and ABTS•+free radical scavenging methods, and in human erythrocytes subjected to oxidative stress with AAPH. In vivo, the antioxidant potential was evaluated inCaenorhabditis elegansnematodes subjected to oxidative stress with juglone. Both cerumen extracts presented phenolic compounds, fatty acids, and metallic minerals in their chemical constitution. The cerumen extracts showed antioxidant activity by capturing free radicals, reducing lipid peroxidation in human erythrocytes, and reducing oxidative stress inC. elegans, observed by the increase in viability. The results obtained indicate that cerumen extracts fromGeotrigonasp. andTetragonisca fiebrigistingless bees may be promising against oxidative stress and associated diseases.