Role of the Hydroxyl Radical-Generating System in the Estimation of the Antioxidant Activity of Plant Extracts by Electron Paramagnetic Resonance (EPR)

Molecules. 2022 Jul 17;27(14):4560. doi: 10.3390/molecules27144560.

Abstract

The scavenging activity of hydroxyl radicals, produced by the Fenton reaction, is commonly used to quantify the antioxidant capacity of plant extracts. In this study, three Fenton systems (Fe/phosphate buffer, Fe/quinolinic acid and Fe/phosphate buffer/quinolinic acid) and the thermal degradation of peroxydisulfate were used to produce hydroxyl radicals; the hydroxyl radical scavenging activity of plant extracts (ginger, blueberry juices and green tea infusion) and chemical compounds (EGCG and GA) was estimated by spin trapping with DMPO (5,5-dimethyl-1-pyrroline N-oxide) and EPR (Electron Paramagnetic Resonance) spectroscopy. Phosphate buffer was used to mimic the physiological pH of cellular systems, while quinolinic acid (pyridine-2,3-dicarboxylic acid) facilitates the experimental procedure by hindering the spontaneous oxidation of Fe(II). The EC50 (the concentration of chemical compounds or plant extracts which halves the intensity of the DMPO-OH adduct) values were determined in all the systems. The results show that, for both the chemical compounds and the plant extracts, there is not a well-defined order for the EC50 values determined in the four hydroxyl radical generating systems. The interactions of phosphate buffer and quinolinic acid with the antioxidants and with potential iron-coordinating ligands present in the plant extracts can justify the observed differences.

Keywords: DMPO; EPR; Fenton reaction; hydroxyl radical; plant extracts.

MeSH terms

  • Antioxidants* / pharmacology
  • Cyclic N-Oxides
  • Electron Spin Resonance Spectroscopy
  • Free Radicals
  • Hydroxyl Radical* / chemistry
  • Phosphates
  • Plant Extracts
  • Quinolinic Acid
  • Spin Labels

Substances

  • Antioxidants
  • Cyclic N-Oxides
  • Free Radicals
  • Phosphates
  • Plant Extracts
  • Spin Labels
  • Hydroxyl Radical
  • Quinolinic Acid

Grants and funding

This research received no external funding.