Direct Assessments of the Antioxidant Effects of the Novel Collagen Peptide on Reactive Oxygen Species Using Electron Spin Resonance Spectroscopy

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In the present study, we evaluated the antioxidant effects of a pepsin-treated novel collagen peptide (P-NCP) on reactive oxygen species (ROS) such as hydroxyl radical (HO^•), superoxide anion radical (O₂^•–), and singlet oxygen (¹O₂), and the effects on cell viability after ultraviolet ray (UV) irradiation of human fibroblasts. We confirmed, using electron spin resonance, that P-NCP directly inhibited HO^• and ¹O₂. Furthermore, addition of P-NCP to fibroblasts inhibited cell death induced by UVA (400 – 315 nm) irradiation in a dose-dependent manner. In addition, the antioxidant effect on ¹O₂ was observed in the peptide fractions rich in Gly, Pro, Hyp, Glu, Ala, and Arg. We found that Gly, Hyp, Glu, and Ala directly scavenged ¹O₂. These results indicated that a peptide sequence including Gly, Hyp, Glu, and Ala could play a key role in the antioxidant effects of P-NCP on ¹O₂. It was suggested that P-NCP can inhibit photo-aging related to ROS owing to its antioxidant effects.