The chemical composition and antimicrobial activity of flavonoids from Sedum aizoon L. against Aeromonas in vitro were investigated, and the effect of flavonoid treatment on the quality of fresh pork during frozen storage for 6 months was also explored. The results showed that kaempferol, quercetin dihydrate, and catechin were the most predominant flavonoids from S. aizoon L. Flavonoids exhibited antibacterial activity to Aeromonas in vitro, which caused membrane damage, disruption of the bacterial surface, and internal ultrastructure, and resulted in the leakage of reducing sugars and proteins. Meanwhile, flavonoid treatment retarded the microbial growth and deteriorates of pork characteristics, including pH value, total volatile basic nitrogen (TVB‐N), texture, and sensory evaluation during frozen storage, thereby prolonged the shelf life. Their results suggested that flavonoids from S. aizoon L. offer a promising choice for food safety and preservation.