Electron Paramagnetic Resonance (EPR) Spectroscopy


The principle of electron paramagnetic resonance (EPR or ESR) is similar to the other forms of spectroscopy which is based on the interaction between the molecule and the light/irradiation (E = hn; DE = gbB) where g is Landé g-factor; beta is the Bohr magneton and B the magnetic field. The EPR covers all the fields of pure and applied sciences (Physics, Chemistry and Geology) but also in biomedical sciences (Biology, Biochemistry). EPR is also applied to medicine field, indicating that it is an accurate and unique tool to evidence the formation or the presence of radical or paramagnetic species within various matrixes. The combination of EPR with chemiluminescence can be useful for detection of reactive oxygen (ROS) or nitrogen (RNS) species.

Recent works have shown that the use of EPR-spin trapping in parallel to High Resolution Oxygraphy (HRO) helps to make correlation between the production of ROS and the mitochondrial dysfunction during anoxia/reoxygenation in cell models and in isolated mitochondria.

At the University of Liège, the Centre of Oxygen, R&D (CORD, Prof. D. Serteyn and Dr. A. Mouithys-Mickalad) has been working for longtime with the Bio-spectroscopy Lab (Prof. M. Hoebeke, Physics Department –Cremant).


EPR technique is used routinely to study:

  • Chemical and biochemical radical processes and their modulation by natural and synthetic molecules
  • Enzyme-catalyzed oxidation of the substrates or probes of interest
  • Anoxia/reoxygenation on primary cells/cell lines and isolated mitochondria
  • Production of ROS by inflammatory cells (human and equine) and their modulation by natural and synthetic compounds
  • Lipid-derived free radicals from polyunsaturated fatty acid (PUFA) using spin trapping technique


EPR applied to Chemistry, Biochemistry and Medicine




EPR applied to Biophysics


Some publications related to the technique:


Shvedova AA, Kisin ER, Murray AR, Mouithys-Mickalad A, Stadler K, Mason RP, Kadiiska M. ESR evidence for in vivo formation of free radicals in tissue of mice exposed to single-walled carbon nanotubes. Free Radic Biol Med. 2014 Aug;73:154-65. doi: 10.1016/j.freeradbiomed.2014.05.010. Epub 2014 May 23. PubMed PMID: 24863695.


Derochette S, Mouithys-Mickalad A, Franck T, Collienne S, Ceusters J, Deby-Dupont G, Neven P, Serteyn D. NDS27 combines the effect of curcumin lysinate and hydroxypropyl-beta-cyclodextrin to inhibit equine PKCδ and NADPH oxidase involved in the oxidative burst of neutrophils. FEBS Open Bio. 2014 Nov 18;4:1021-9. doi: 10.1016/j.fob.2014.11.004. eCollection 2014. PubMed PMID: 25493216; PubMed Central PMCID: PMC4254746.


Tsumbu CN, Deby-Dupont G, Tits M, Angenot L, Franck T, Serteyn D, Mouithys-Mickalad A. Antioxidant and antiradical activities of Manihot esculenta  Crantz (Euphorbiaceae) leaves and other selected tropical green vegetables investigated on lipoperoxidation and phorbol-12-myristate-13-acetate (PMA) activated monocytes. Nutrients. 2011 Sep;3(9):818-38. doi: 10.3390/nu3090818. Epub 2011 Sep 16. PubMed PMID: 22254126; PubMed Central PMCID: PMC3257738.


Sandersen C, Mouithys-Mickalad A, de la Rebière G, Deby G, Serteyn D, Franck T. Modulating effects of acepromazine on the reactive oxygen species production by stimulated equine neutrophils. Vet Anaesth Analg. 2011 Mar;38(2):83-93. doi: 10.1111/j.1467-2995.2010.00583.x. PubMed PMID: 21303439.


de Rebière de Pouyade G, Salciccia A, Ceusters J, Deby-Dupont G, Serteyn D, Mouithys-Mickalad A. Production of free radicals and oxygen consumption by primary equine endothelial cells during anoxia-reoxygenation. Open Biochem J. 2011;5:52-9. doi: 10.2174/1874091X01105010052. Epub 2011 Nov 24. PubMed PMID: 22207886; PubMed Central PMCID: PMC3242399.


Grammenos A, Mouithys-Mickalad A, Guelluy PH, Lismont M, Piel G, Hoebeke M. ESR technique for noninvasive way to quantify cyclodextrins effect on cell membranes. Biochem Biophys Res Commun. 2010 Jul 30;398(3):350-4. doi: 10.1016/j.bbrc.2010.06.050. Epub 2010 Jun 17. PubMed PMID: 20599748.


Chignell CF, Han SK, Mouithys-Mickalad A, Sik RH, Stadler K, Kadiiska MB. EPR studies of in vivo radical production by 3,3',5,5'-tetrabromobisphenol A (TBBPA) in the Sprague-Dawley rat. Toxicol Appl Pharmacol. 2008 Jul 1;230(1):17-22. doi: 10.1016/j.taap.2008.01.035. Epub 2008 Feb 14. PubMed PMID: 18342900.


Navet R, Mouithys-Mickalad A, Douette P, Sluse-Goffart CM, Jarmuszkiewicz W, Sluse FE. Proton leak induced by reactive oxygen species produced during in vitro anoxia/reoxygenation in rat skeletal muscle mitochondria. J Bioenerg Biomembr. 2006 Feb;38(1):23-32. PubMed PMID: 16733628.


Schneider N, Mouithys-Mickalad AL, Lejeune JP, Deby-Dupont GP, Hoebeke M, Serteyn DA. Synoviocytes, not chondrocytes, release free radicals after cycles of anoxia/re-oxygenation. Biochem Biophys Res Commun. 2005 Aug 26;334(2):669-73. PubMed PMID: 16036131.


Bahri MA, Heyne BJ, Hans P, Seret AE, Mouithys-Mickalad AA, Hoebeke MD. Quantification of lipid bilayer effective microviscosity and fluidity effect induced by propofol. Biophys Chem. 2005 Apr 1;114(1):53-61. Epub 2004 Nov 21. PubMed PMID: 15792861.


Kohnen S, Halusiak E, Mouithys-Mickalad A, Deby-Dupont G, Deby C, Hans P, Lamy M, Noels AF. Catalytic activation of copper (II) salts on the reaction of peroxynitrite with propofol in alkaline medium. Nitric Oxide. 2005 Jun;12(4):252-60. Epub 2005 Apr 19. PubMed PMID: 15917218.


Mouithys-Mickalad A, Deby-Dupont G, Dogne JM, de Leval X, Kohnen S, Navet R, Sluse F, Hoebeke M, Pirotte B, Lamy M. Effects of COX-2 inhibitors on ROS produced by Chlamydia pneumoniae-primed human promonocytic cells (THP-1). Biochem Biophys Res Commun. 2004 Dec 24;325(4):1122-30. PubMed PMID: 15555544.


Mouithys-Mickalad A, Deby-Dupont G, Mathy-Hartert M, Habraken Y, Nys M, Henrotin Y, Lamy M, Deby C. Effects of glucocorticoids on the respiratory burst of Chlamydia-primed THP-1 cells. Biochem Biophys Res Commun. 2004 Jun 11;318(4):941-8. PubMed PMID: 15147963.


Kohnen SL, Mouithys-Mickalad AA, Deby-Dupont GP, Deby CM, Hans P, Lamy ML, Noels AF. Investigation of the reaction of peroxynitrite with propofol at acid pH: predominant production of oxidized, nitrated, and halogenated derivatives. Nitric Oxide. 2003 May;8(3):170-81. PubMed PMID: 12826066.


Mouithys-Mickalad A, Mathy-Hartert M, Du G, Sluse F, Deby C, Lamy M, Deby-Dupont G. Oxygen consumption and electron spin resonance studies of free radical production by alveolar cells exposed to anoxia: inhibiting effects of the antibiotic ceftazidime. Redox Rep. 2002;7(2):85-94. PubMed PMID: 12189054.


Kohnen SL, Mouithys-Mickalad AA, Deby-Dupont GP, Deby CM, Lamy ML, Noels AF.Oxidation of tetrahydrobiopterin by peroxynitrite or oxoferryl species occurs by a radical pathway. Free Radic Res. 2001 Dec;35(6):709-21. PubMed PMID: 11811523.


Mouithys-Mickalad AM, Zheng SX, Deby-Dupont GP, Deby CM, Lamy MM, Reginster JY, Henrotin YE. In vitro study of the antioxidant properties of non steroidal anti-inflammatory drugs by chemiluminescence and electron spin resonance (ESR). Free Radic Res. 2000 Nov;33(5):607-21. PubMed PMID: 11200092.


Du G, Willet K, Mouithys-Mickalad A, Sluse-Goffart CM, Droy-Lefaix MT, Sluse FE. EGb 761 protects liver mitochondria against injury induced by in vitro anoxia/reoxygenation. Free Radic Biol Med. 1999 Sep;27(5-6):596-604. PubMed PMID:10490280.


Benbarek H, Mouithys-Mickalad A, Deby-Dupont G, Deby C, Grülke S, Nemmar A, Lamy M, Serteyn D. High concentrations of histamine stimulate equine polymorphonuclear neutrophils to produce reactive oxygen species. Inflamm Res. 1999 Nov;48(11):594-601. PubMed PMID: 10598016.


Liègeois JF, Bruhwyler J, Petit C, Damas J, Delarge J, Gèczy J, Kauffmann JM, Lamy M, Meltzer H, Mouithys-Mickalad A. Oxidation sensitivity may be a useful tool for the detection of the hematotoxic potential of newly developed molecules: application to antipsychotic drugs. Arch Biochem Biophys. 1999 Oct 1;370(1):126-37. PubMed PMID: 10496986.


Du G, Mouithys-Mickalad A, Sluse FE. Generation of superoxide anion by mitochondria and impairment of their functions during anoxia and reoxygenation in vitro. Free Radic Biol Med. 1998 Dec;25(9):1066-74. PubMed PMID: 9870560.


Mouithys-Mickalad A, Hans P, Deby-Dupont G, Hoebeke M, Deby C, Lamy M. Propofol reacts with peroxynitrite to form a phenoxyl radical: demonstration by electron spin resonance. Biochem Biophys Res Commun. 1998 Aug 28;249(3):833-7. PubMed PMID: 9731222.


Mouithys-Mickalad A, Deby C, Deby-Dupont G, Lamy M. An electron spin resonance (ESR) study on the mechanism of ascorbyl radical production by metal-binding proteins. Biometals. 1998 Apr;11(2):81-8. PubMed PMID: 9542060.


Mouithys-Mickalad A, Deby-Dupont G, Hoebeke M, Mathy-Hartert M, Lamy M, Deby C. Effects of sphingosine and sphingosine analogues on the free radical production by stimulated neutrophils: ESR and chemiluminescence studies. Mediators Inflamm. 1997;6(5-6):327-33. PubMed PMID: 18472867; PubMed Central PMCID: PMC2365874.


Liègeois JF, Mouithys-Mickalad A, Bruhwyler J, Delarge J, Petit C, Kauffmann JM, Lamy M. JL 13, a potential successor to clozapine, is less sensitive to oxidative phenomena. Biochem Biophys Res Commun. 1997 Sep 8;238(1):252-5. PubMed PMID: 9299488.