Previous work showed that peptides from duck eggs incubated for 15 D presented high total antioxidant activities. Here, this work explore the antioxidant activities of different segments, ZT1 (≤3 KD), ZT2 (≤10 KD), and ZT3 (≤30 KD), derived from duck embryo peptides and their protective effects against H2O2-induced oxidative damage in HepG2 cells. Peptides present no cytotoxicity to HepG2 cells. Moreover, ZT1 exhibits a higher ability to scavenge several radicals as well as stronger inhibition of H2O2-induced oxidative stress than ZT2 and ZT3. The activities of catalase and glutathione peroxidase as well as total superoxide dismutase increase in a concentration-dependent manner. Peptides are isolated from ZT1 and then subjected to LC-MS/MS to identify their sequences, followed by functional annotation, bioinformatics prediction, and hot-spot motif recognition. As a result, 413 potential functional peptides are identified, with some compounds exhibiting more than 1 function. This work will help for exploring bioactive substances in duck embryo eggs and enhance the utilization value of duck or other poultry eggs.

1. Oxidative stress in duck embryos undergoes dynamic change during incubation. However, the detailed change characteristics has not been studied yet. Here, we explored the dynamic change characteristics of different antioxidant substances in duck embryo eggs during incubation. 2. The following trial assayed antioxidant substances, including vitamins E (VE) and C (VC), glutathione (GSH), reduced glutathione (reduced GSH), oxidative glutathione (GSSG), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-px), and malonic dialdehyde (MDA). Five antioxidant peptides (DY1, DY2, DY3, DY4 and DY5) were screened and their protective effect on HepG2 cells and their mechanism of action determined. 3. The results showed that antioxidant substances underwent a dynamic change during incubation through various mechanisms. Moreover, the T-AOC of duck embryos at different incubation times, firstly increased and then decreased, reaching the highest level on d 15 during incubation. The peptide DY2 (TVDGPSGKLWRD) exhibited high antioxidant activity in vitro, and is known to regulate the apoptosis pathway in mitochondria. 4. The data indicated that duck embryos can be used for the development of related antioxidant products and purification of new bioactive peptides.