Methods to assess free radicals and oxidative stress in biological systems.
Oxidative stress results from a disruption of the prooxidant/antioxidant cellular balance and monitoring free radical status becomes an interesting challenge in animal and human nutrition. In the present work, merits and limitations of different analytical techniques (HPLC, GC-MS, fluorometric and colourometric assays, ELISA, gel electrophoresis) for the measurement of radical mediated alterations in the cellular integrity of lipids (malondialdehyde, hydrocarbon gases, F2-isoprostanes) proteins (protein carbonyls, 3-nitrotyrosine) and DNA (8-hydroxy-2'-deoxyguanosine) are discussed. Besides these indirect methods, owing to the fact that free radicals are paramagnetic, electron paramagnetic resonance spectroscopy combined with spin trapping has become a valuable tool to directly assess and to better understand the mechanisms of free radical reactions. With this approach a radical that is too short-lived to be detected, adds to a spin-trapping agent to form a relatively long-lived radical adduct. Information obtained from the hyperfine splitting of the spin-trapped adduct can provide identification and quantification of the originally generated free radicals.
Rimbach
G
G
Department of Molecular and Cell Biology, University of California at Berkeley 94720-3200, USA. rimbach@hotmail.com
Höhler
D
D
Fischer
A
A
Roy
S
S
Virgili
F
F
Pallauf
J
J
Packer
L
L
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003124:Colorimetry; D004249:DNA Damage; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D004586:Electrophoresis; D004797:Enzyme-Linked Immunosorbent Assay; D005470:Fluorometry; D005609:Free Radicals / Q000032:analysis*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D015227:Lipid Peroxidation; D018384:Oxidative Stress; D011506:Proteins / Q000378:metabolism; D019097:Spin Trapping
D005609:Free Radicals; D011506:Proteins