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Related Experiment Videos

Radicals from "Good's" buffers.

J K Grady1, N D Chasteen, D C Harris

  • 1Department of Chemistry, University of New Hampshire, Durham 03824-3598.

Analytical Biochemistry
|August 15, 1988
PubMed
Summary

Iron-catalyzed reactions can generate harmful radicals from piperazine-based Good

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Area of Science:

  • Biochemistry
  • Chemical Biology
  • Free Radical Chemistry

Background:

  • Good's buffers are widely used in biochemical research.
  • Iron-mediated reactions can generate reactive oxygen species.
  • The behavior of buffers under oxidative stress is not fully understood.

Purpose of the Study:

  • To investigate radical formation from Good's buffers during iron-catalyzed oxidative processes.
  • To identify the specific buffer components involved in radical generation.
  • To assess the implications for redox studies in biochemistry.

Main Methods:

  • Electron Paramagnetic Resonance (EPR) spectroscopy to detect and characterize radical species.
  • Assays for hydroxyl radical production (deoxyribose/thiobarbituric acid).
  • Electrochemical methods to induce radical formation.
  • Enzymatic studies using superoxide dismutase and catalase.

Main Results:

  • Piperazine-ring buffers (Hepes, Epps, Pipes) form stable radicals with ~10 min half-lives.
  • Morpholine-ring buffer (Mes) does not form radicals.
  • Buffer radicals are secondary species derived from oxygen radicals in the Haber-Weiss process.
  • Hepes acts as a hydroxyl radical scavenger and forms a nitroxide radical under specific conditions.

Conclusions:

  • Piperazine-containing Good's buffers are susceptible to radical formation in the presence of iron and oxidative conditions.
  • These buffer-derived radicals can interfere with redox-sensitive biochemical experiments.
  • Researchers should avoid piperazine-based buffers in studies involving iron redox chemistry.

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