Hylocereus undatus flower is commonly used as food or for medicinal purposes in south China. To study its antioxidant activity and mechanism we used antioxidant and chemical assays to compare two commercial samples from different locations (Shenjing, Qixing). The difference in antioxidant levels corresponded with differences in chemical content (including total phenolics, total flavonoids, kaempferol and quercetin) between Shenjing and Qixing. The antioxidant ability of H. undatus flower seems attributable to total phenolics (mainly total flavonoids). Kaempferol is one of the main bioactive components. H. undatus flower exerts its antioxidant effects through metal chelation and radical scavenging via hydrogen atom (H•) and electron (e) donation.