EPR in Chemistry

Enzyme Reactions

Detection and study of the active site of Cu,Zn-SOD
Many enzyme reactions involve one-electron oxidation steps with formation of paramagnetic transient state of the enzyme detectable by EPR. The paramagnetic center where the unpaired electron is located, is usually centered at a transition metal (metalloproteins) or is an amino acid derived radical. Detection and identification of the paramagnetic centers is important to understand the function of the enzymes. For example, in the native SOD1 enzyme, the active site contains one Cu(II) ion that gives a very characteristic EPR spectrum.

Crystal structure (1E9P.pdb) of Cu(II)-SOD protein
Cu(II)-SOD蛋白在77 K的EPR光谱使用手指Dewar

Reaction Kinetics

Kinetics analysis of vitamin C antioxidant ability
Many chemical reactions involve the transfer of one electron. Each electron transfer results in an unpaired electron creating paramagnetic free radicals. EPR is the ideal spectroscopic technique to measure these species as well as to monitor the time behavior of their creation and disappearance. EPR solely has the ability to detect free radicals unambiguously. For example, antioxidants such as vitamin C are important in neutralizing dangerous free radicals in living things and kinetics indicates their effectiveness.

Experimental data of the reduction of the nitroxide TEMPOL by ascorbate (vitamin C)
Mechanism of TEMPOL reduction by vitamin C

Photochemistry

Light degradation of hops in beer
大多数光化学反应是通过自由基形成中间体进行的。例如,在酿造工艺中使用的啤酒花包含包括humulones,sumumulones,adhumulones,beta酸和精油在内的活性成分的混合物。这些组件的某些形式是光活性的。啤酒的轻曝光导致形成自由基,与硫化合物结合在一起,并为啤酒带来令人不愉快的风味和气味。

使用UV灯配件将HOP产品暴露于220-600 nm的UV/VIS灯。在旋转陷阱DMPO的存在下记录了光诱导的自由基,并鉴定为超氧化物阴离子自由基和两个以C为中心的自由基。

Catalysis

Hydroxyl radical generation through photocatalytic reaction of TiO2
The modern chemical industry relies heavily on homogeneous and heterogeneous catalysts. Understanding the operational mode, or reactivity, of these catalysts is crucial for improved developments and enhanced performance. Where paramagnetic centers are involved, ranging from transition metal ions to defects and radicals, EPR spectroscopy is without doubt the technique of choice. For example, photocatalytic oxidation of organic pollutants is frequently carried out using semiconducting polycrystalline powders such as TiO2. A hydroxyl radical is easily formed by light irradiation of TiO2 and detected by EPR using spin traps.

Mechanism of hydroxyl radical formation upon light irradiation of TiO2
EPR spectra obtained upon irradiation of aqueous TiO2 suspensions in the presence of spin trapping agent PBN

Electrochemistry

EPR electrochemistry study of ruthenium complexes
Electrochemical generation method combined with EPR has been used to identify and investigate free radicals derived from both organic and inorganic compounds. Inorganic dyes can be used to improve the efficiency of solar cells. In order to optimize the ligands, one must understand the electronic structure of the dye. Here the electrochemistry and EPR combined with DFT calculations and UV/Vis spectroscopy show the unpaired electron is delocalized between the metal and the ligand.

Data courtesy of Prof. J. Rochford, University of Massachusetts Boston (Inorg. Chem., 2016, 55 (5), pp 2460–2472)

Redox Chemistry

通过Cu(II)还原研究的SOD蛋白的酶促活性
Enzymes in the human body regulate oxidation-reduction reactions. These complex proteins, of which several hundred are known, act as catalysts, speeding up chemical processes in the body. Oxidation-reduction reactions also take place in the metabolism of food for energy, with substances in the food broken down into components the body can use. For example, the dismutase activity of Cu,Zn-SOD protein involves reduction of Cu(II)-SOD to Cu(I)-SOD:

减少铜(II)草皮活跃(EPR)铜(I)草皮(EPR inactive)
Cu(II)-SOD has a very characteristic EPR signal which decays upon reduction of Cu(II) -> Cu(I).

Antioxidants

Detection of ascorbate radical upon oxidation of vitamin C
自由基的有利影响和有害影响之间的微妙平衡是人(Patho)生理学的重要方面之一。有毒自由基的产生不平衡与许多疾病的发病机理高度相关,这些疾病需要应用选定的抗氧化剂来恢复体内平衡。EPR用于使用内源性长寿命的自由基(Ascorbyl Ladical,Docopheloxyl自由基,黑色素)确定生物系统的氧化状态。

Reaction of a toxic radical R● with an antioxidant A. Also pictured is the reaction of the antioxidant ascorbic acid (Vitamin C) with a radical
EPR spectrum of ascorbate (Vitamin C) radical