An on-line post-column detection system for the detection of reactive-oxygen-species-producing compounds and antioxidants in mixtures |
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Authors: | Jeroen Kool Sebastiaan M Van Liempd Stefan Harmsen Tim Schenk Hubertus Irth Jan N M Commandeur Nico P E Vermeulen |
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Institution: | (1) LACDR-Division of Molecular Toxicology, Department of Pharmacochemistry, Vrije Universiteit, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands;(2) Kiadis B.V., 9747 AN Groningen, The Netherlands |
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Abstract: | 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. |
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Keywords: | Pro-oxidant Antioxidant ROS Bioactivation On-line Biochemical detection |
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