Publication Date

9-8-2025

Document Type

Tutorial Material

Keywords

Electron paramagnetic resonance (EPR), Laboratory experiment, Undergraduate level

Abstract

We report a biochemically based EPR experiment for undergraduate laboratories that will serve as instruction for learning EPR sample preparation, spectral observation, and spectral analysis. There is a current lack of EPR instructional experiments that present complex spectra in which computational analysis may be required. The biochemical oxidation system of horse radish peroxidase (HRP) activated by H2O2 acts on 2,6-di-t-butyl-4-hydroxyanisole (DTBHA) to form phenoxyl radicals. This experiment produces moderately complex spectra of moderately long-lived radicals using relatively small amounts of readily available reactants. Spectrum analysis and simulation is accomplished using software freely available on-line. If desired, EPR studies may be extended to related systems such as the food additives butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) with shorter-lived radicals. If appropriate, molecular orbital calculations at various levels of sophistication may be employed to rationalize EPR results.

Copyright Date

9-1-2025

Rights Holder

Evan A. Old, Matthew C. Rehak, Jefferson C. Thompson, and Herbert J. Sipe, Jr.

Copyright Statement / License for Reuse

All Rights Reserved
All Rights Reserved.

Organizational Units

College of Natural Science and Mathematics, Chemistry and Biochemistry

Provenance

Received from author

File Format

application/pdf

Language

English (eng)

Extent

16 pgs

File Size

2.9 MB

Publication Statement

Copyright is held by the authors. User is responsible for all copyright compliance.

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