Abstract
Free radicals including reactive oxygen species (ROS) are important chemicals in the research area of biology, pharmaceutical, medical, and environmental science as well as human health risk assessment as they are highly involved in diverse metabolism and toxicity mechanisms through chemical reactions with various components of living bodies. Electron spin resonance (ESR) spectroscopy is a powerful tool for detecting and quantifying those radicals in biological environments. In this work we observed the ESR signal of 2,2,6,6-Tetra-methyl piperidine 1-oxyl (TEMPO) in aqueous solution at various concentrations to estimate the uncertainty factors arising from the experimental conditions and signal treatment methods. As the sample position highly influences the signal intensity, dual ESR tube geometry (consists of a detachable sample tube and a position fixed external tube) was adopted. This type of measurement geometry allowed to get the relative uncertainty of signal intensity lower than 1% when triple measurements are averaged. Linear dependence of signal intensity on the TEMPO concentration, which is required for the quantification of unknown sample, could be obtained over a concentration range of ~103 by optimizing the signal treatment method depending on the concentration range.