Quantification of antioxidant activity in brain tissue homogenates using the 'total equivalent antioxidant capacity'.
We demonstrate herein that the standard 'Trolox equivalent antioxidant capacity' (TEAC) assay, typically utilized to quantify total antioxidant levels within plasma, can also be utilized for tissue homogenates. Normal mice and transgenic mice lacking apolipoprotein E were subjected to a diet including iron as a generic pro-oxidant for 1 month (which has been shown to induce oxidative damage in our prior studies) and homogenates of brain tissue were subjected to the TEAC assay. Levels of the endogenous antioxidant glutathione levels were also monitored by HPLC. As described previously, ApoE-deficient mice expressed increased levels of glutathione; total antioxidant levels, as determined by TEAC, were increased to a similar extent. The increase in total antioxidant levels, as determined by TEAC, following dietary iron challenge paralleled the increase in glutathione levels, as determined by HPLC. These findings indicate that the TEAC assay may be useful for tissue homogenates. The rapid nature of this assay compared to HPLC, coupled with its lack of requirement for sophisticated equipment, makes it well suited for analyses of multiple tissue samples.
Shea
Thomas B
TB
Center for Cellular Neurobiology, University of Massachusetts-Lowell, 01854, Lowell, MA, USA. thomas_shea@uml.edu
Rogers
Eugene
E
Ashline
David
D
Ortiz
Daniela
D
Sheu
Min Shyan
MS
D000818:Animals; D000975:Antioxidants / Q000032:analysis*; D001057:Apolipoproteins E / Q000235:genetics*; D001921:Brain / Q000378:metabolism*; D001923:Brain Chemistry; D002839:Chromans / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D004032:Diet; D005978:Glutathione / Q000032:analysis*; D007501:Iron / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000032:analysis
D000975:Antioxidants; D001057:Apolipoproteins E; D002839:Chromans; D012709:Serum Albumin; D007501:Iron; D005978:Glutathione; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid