Jump to Main Content
An on-line post-column detection system for the detection of reactive-oxygen-species-producing compounds and antioxidants in mixtures
- Kool, Jeroen, Van Liempd, Sebastiaan M., Harmsen, Stefan, Schenk, Tim, Irth, Hubertus, Commandeur, Jan N. M., Vermeulen, Nico P. E.
- Analytical and bioanalytical chemistry 2007 v.388 no.4 pp. 871-879
- DNA, antioxidant activity, antioxidants, cytochrome P-450, high performance liquid chromatography, lipid peroxidation, neutralization, reactive oxygen species, screening
- Reactive oxygen species (ROS) can damage proteins, cause lipid peroxidation, and react with DNA, ultimately resulting in harmful effects. Antioxidants constitute one of the defense systems used to neutralize pro-oxidants. Since pro-oxidants and antioxidants are found ubiquitously in nature, pro-and antioxidant effects of individual compounds and of mixtures receive much attention in scientific research. A major bottleneck in these studies, however, is the identification of the individual pro-oxidants and antioxidants in mixtures. Here, we describe the development and validation of an on-line post-column biochemical detection system for ROS-producing compounds and antioxidants in mixtures. Inclusion of cytochrome P450s and cytochrome P450 reductase also permitted the screening of compounds that need bioactivation to exert their ROS-producing properties. This pro-oxidant and antioxidant detection system was integrated on-line with gradient HPLC. The resulting high-resolution screening technology was able to separate mixtures of ROS-producing compounds and antioxidants, allowing each species to be characterized rapidly and sensitively.