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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
Summary
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.
- 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.