A new method is proposed for measuring the antioxidant capacity by electron spin resonance spectroscopy based on the loss of electron spin resonance signal after Cu(2+) is reduced to Cu(+) with antioxidant. Cu(+) was removed by precipitation in the presence of SCN(-). The remaining Cu(2+) was coordinated with diethyldithiocarbamate, extracted into n-butanol and determined by electron spin resonance spectrometry. Eight standards widely used in antioxidant capacity determination, including Trolox, ascorbic acid, ferulic acid, rutin, caffeic acid, quercetin, chlorogenic acid, and gallic acid were investigated. The standard curves for determining the eight standards were plotted, and results showed that the linear regression correlation coefficients were all high enough (r > 0.99). Trolox equivalent antioxidant capacity values for the antioxidant standards were calculated, and a good correlation (r > 0.94) between the values obtained by the present method and cupric reducing antioxidant capacity method was observed. The present method was applied to the analysis of real fruit samples and the evaluation of the antioxidant capacity of these fruits.
Electron spin resonance spectroscopy was applied to determining the antioxidant capacity of eight thiol-containing compounds, including reduced glutathione, N-acetyl-L-cycsteine, methimazole, captopril, and tiopronin with one thiol group, 1,4-dithioerythritol and 2,3-dimercapto-1-propanol with two thiol groups, as well as L-cystine with no free thiol group. Cu ion gives an electron spin resonance signal and is reduced to Cu ion with no electron spin resonance signal by the free thiol group in the compounds. Trolox equivalent antioxidant capacity (TEAC) was used to evaluate the reducing ability of the thiol-containing compounds and the TEAC values were found to be relevant to the number of thiol groups contained in the compounds. For the purpose of comparison, the UV-vis spectrophotometry, cupric reducing antioxidant capacity (CUPRAC) method, and Ellman assay were applied to the determination of the antioxidant capacity of the thiol-containing compounds. The TEAC values obtained by the present method were very close to those obtained by UV-vis method. However, compared with CUPRAC method, for methimazole the present method gave a more reasonable TEAC value. The present method was also applied to the quantification of N-acetyl-L-cycsteine, methimazole, captopril, and tiopronin in their pharmaceutical formulations.
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