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Related Experiment Videos

Iron speciation at physiological pH in media containing ascorbate and oxygen

C Dorey1, C Cooper, D P Dickson

  • 1Division of Clinical Cell Biology, MRC Clinical Research Centre, Harrow, Middlesex.

The British Journal of Nutrition
|July 1, 1993
PubMed
Summary

Iron ascorbate solutions are unstable at physiological pH, rapidly forming iron(III) species and consuming oxygen. Mannitol can partially stabilize these iron solutions, impacting iron physiology studies.

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Area of Science:

  • Biochemistry
  • Inorganic Chemistry
  • Physiology

Background:

  • Iron ascorbate complexes are utilized in various physiological studies.
  • Understanding the stability of these solutions is crucial for accurate experimental outcomes.
  • Previous research has not fully elucidated the speciation and stability of iron ascorbate at physiological pH.

Purpose of the Study:

  • To investigate the stability of iron ascorbate solutions under varying pH and oxygen conditions.
  • To characterize the iron species formed in these solutions using spectroscopic and chromatographic methods.
  • To assess the implications of iron ascorbate instability on iron physiology research.

Main Methods:

  • Spectroscopic analysis including Electron Paramagnetic Resonance (EPR) and Mössbauer spectroscopy.

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  • Indicator studies using ferrozine (Fe2+) and mimosine (Fe3+).
  • Gel-filtration chromatography and oxygen uptake measurements.

    • Main Results:

    • Iron ascorbate solutions rapidly form mononuclear, chelatable iron(III) species at physiological pH (6.8-7.4).
    • Mössbauer spectroscopy detected no Fe2+ species, while EPR showed a decrease in Fe3+ over time, especially in oxygenated buffers, indicating polynuclear iron formation.
    • Oxygen consumption correlated with Fe2+ to Fe3+ conversion above pH 7.0, preceding ascorbate depletion. Mannitol partially stabilized the solutions.

    Conclusions:

    • Iron ascorbate solutions exhibit inherent instability above neutral pH.
    • The formation of polynuclear iron species and rapid oxygen consumption are key features of this instability.
    • These findings have significant implications for the interpretation of results in iron physiology studies using ascorbate.