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A new method to measure nitrate/nitrite with a NO-sensitive electrode.

R Berkels1, S Purol-Schnabel, R Roesen

  • 1Institut fuer Pharmakologie, Universitaet zu Koeln, 50931 Koeln, Germany. Reinhard.Berkels@medizin.uni-koeln.de

Journal of Applied Physiology (Bethesda, Md. : 1985)
|January 3, 2001
PubMed
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Researchers developed a sensitive new method to measure nitric oxide (NO) by converting its degradation products, nitrate and nitrite, back to NO. This technique allows for precise measurement in biological systems using a standard electrode.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Physiology

Background:

  • Nitric oxide (NO) is a crucial signaling molecule.
  • Measuring NO in biological systems is challenging due to its instability.
  • Nitrate and nitrite are stable degradation products of NO.

Purpose of the Study:

  • To develop a novel, highly sensitive method for quantifying nitric oxide (NO).
  • To enable the measurement of NO and its stable metabolites, nitrate and nitrite.
  • To assess basal and stimulated NO release in various biological models.

Main Methods:

  • Enzymatic conversion of nitrate to nitrite using nitrate reductase.
  • Acidic iodide reduction of nitrite to equimolar concentrations of NO.
  • Detection of NO using a commercially available amperometric Clark-type electrode.

Related Experiment Videos

  • Quantification of NO in aqueous solutions with a detection limit of 2 nmol/l.

    • Main Results:

    • Demonstrated a sensitive method for NO measurement via nitrate/nitrite reconversion.
    • Successfully measured basal and carbachol-stimulated NO release from endothelial cells.
    • Quantified nitrate/nitrite concentrations in guinea pig coronary effluent.
    • Achieved a detection limit of 2 nmol/l NO.

    Conclusions:

    • Presents a valid and highly sensitive method for measuring nitrite/NO in biological samples.
    • The developed technique is applicable to diverse biological models, including cell cultures and in vivo samples.
    • Utilizes readily available commercial electrodes for practical application in research.