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Related Experiment Videos

A polarographic method for measuring dissolved nitric oxide

B O Jensen1, J Skeidsvoll, H Holmsen

  • 1Department of Biochemistry and Molecular Biology, University of Bergen, Norway.

Journal of Biochemical and Biophysical Methods
|February 21, 1998
PubMed
Summary
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A new polarographic method accurately measures nitric oxide (NO) concentration in solutions by quantifying consumed oxygen. This simple technique simplifies NO solution preparation and storage for biological studies.

Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Physiology

Background:

  • Nitric oxide (NO) is a crucial signaling molecule in biological systems.
  • Accurate measurement of NO concentration is essential for understanding its physiological roles.
  • Existing methods for NO quantification can be complex or require specific calibration standards.

Purpose of the Study:

  • To develop and validate a simple polarographic method for measuring authentic nitric oxide (NO) concentration in aqueous solutions.
  • To establish a reliable method for quantifying NO that is suitable for immediate use and long-term storage of NO solutions.
  • To demonstrate the utility of this method for studying biological effects of NO.

Main Methods:

  • A polarographic technique was employed to measure oxygen consumption during the reaction of NO with dissolved oxygen.

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  • Nitrite formation, a product of the NO-oxygen reaction, was analyzed using capillary electrophoresis.
  • A linear correlation (R2 = 0.89) was established between oxygen consumed and nitrite formed (20-100 nmol NO).

    • Main Results:

    • A direct, linear relationship was observed between oxygen consumption and nitrite formation, confirming the reaction stoichiometry.
    • The polarographic method accurately estimates dissolved NO concentrations in authentic solutions.
    • The method demonstrated simplicity, immediate applicability, and independence from NO calibration standards.

    Conclusions:

    • Polarographic measurement of oxygen consumption provides a reliable and straightforward method for quantifying NO in aqueous solutions.
    • This technique simplifies the preparation, storage, and measurement of NO solutions, facilitating research.
    • The method enables in vitro studies of NO's rapid biological effects through controlled infusion into cell suspensions.