A hydroxyl-scavenging assay based on DNA damage in vitro.

Previous deoxyribose degradation method lacks biological relevance, specificity, and even reliability. In this study, a new hydroxyl radical-scavenging assay based on DNA damage is described. 2-Thiobarbituric acid-reactive substance (TBARS, λmax = 530 nm) was chosen as the biomarker of hydroxyl-mediated DNA damage. On the basis of systematic investigations into various factors affecting A530 nm and solvent interference, the experimental procedure was developed. The successful measurement of 30 selected antioxidants demonstrated that the proposed DNA damage method is reliable, simple, specific, and biologically relevant. It is suitable for all types of antioxidants in vitro.