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Review
. 2018 May 20;28(15):1433-1443.
doi: 10.1089/ars.2017.7396. Epub 2017 Nov 17.

Electron Paramagnetic Resonance Measurements of Reactive Oxygen Species by Cyclic Hydroxylamine Spin Probes

Sergey I Dikalov  1 , Yuliya F Polienko  2   3 , Igor Kirilyuk  2   3
Affiliations
Review

Electron Paramagnetic Resonance Measurements of Reactive Oxygen Species by Cyclic Hydroxylamine Spin Probes

Sergey I Dikalov et al. Antioxid Redox Signal. .

Abstract

Significance: Oxidative stress contributes to numerous pathophysiological conditions such as development of cancer, neurodegenerative, and cardiovascular diseases. A variety of measurements of oxidative stress markers in biological systems have been developed; however, many of these methods are not specific, can produce artifacts, and do not directly detect the free radicals and reactive oxygen species (ROS) that cause oxidative stress. Electron paramagnetic resonance (EPR) is a unique tool that allows direct measurements of free radical species. Cyclic hydroxylamines are useful and convenient molecular probes that readily react with ROS to produce stable nitroxide radicals, which can be quantitatively measured by EPR. In this work, we critically review recent applications of various cyclic hydroxylamine spin probes in biology to study oxidative stress, their advantages, and the shortcomings. Recent Advances: In the past decade, a number of new cyclic hydroxylamine spin probes have been developed and their successful application for ROS measurement using EPR has been published. These new state-of-the-art methods provide improved selectivity and sensitivity for in vitro and in vivo studies.

Critical issues: Although cyclic hydroxylamine spin probes EPR application has been previously described, there has been lack of translation of these new methods into biomedical research, limiting their widespread use. This work summarizes "best practice" in applications of cyclic hydroxylamine spin probes to assist with EPR studies of oxidative stress.

Future directions: Additional studies to advance hydroxylamine spin probes from the "basic science" to biomedical applications are needed and could lead to better understanding of pathological conditions associated with oxidative stress. Antioxid. Redox Signal. 28, 1433-1443.

Keywords: electron paramagnetic resonance; electron spin resonance; hydroxylamine spin probes; reactive oxygen species; superoxide.

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Figures

<b>FIG. 1.</b>
FIG. 1.
Chemical structures of hydroxylamine spin probes.
<b>FIG. 2.</b>
FIG. 2.
Reactions of cyclic hydroxylamines with ROS. *The constant for reaction of TEMPONE-H with peroxynitrite is given (17). ROS, reactive oxygen species.
<b>FIG. 3.</b>
FIG. 3.
Interconversion of cyclic hydroxylamines and nitroxides in superoxide-generating systems.
<b>FIG. 4.</b>
FIG. 4.
Preparation of the frozen tissue for ROS measurements.
<b>FIG. 5.</b>
FIG. 5.
Preparation of the frozen samples with the blood or cell suspension.
<b>FIG. 6.</b>
FIG. 6.
Acyl-protected hydroxylamine probes.
See this image and copyright information in PMC

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