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Related Experiment Videos

Nitrite and nitrate determinations in plasma: a critical evaluation

H Moshage1, B Kok, J R Huizenga

  • 1Department of Medicine, University Hospital Groningen, The Netherlands.

Clinical Chemistry
|June 1, 1995
PubMed
Summary

Plasma nitrite and nitrate measurements are valuable clinical markers. However, whole blood nitrite rapidly converts to nitrate, making plasma nitrite determination alone unreliable. Proper sample preparation is crucial to avoid artifacts.

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Area of Science:

  • Clinical chemistry
  • Biochemical analysis
  • Nitric oxide research

Background:

  • Plasma nitrite and nitrate are increasingly used as biomarkers for nitric oxide synthase activity and nitric oxide radical production.
  • A systematic evaluation of plasma nitrite and nitrate determination methods is lacking.

Purpose of the Study:

  • To systematically evaluate the determination of nitrite and nitrate in plasma.
  • To assess recovery, stability, and potential artifact sources in plasma and whole blood samples.

Main Methods:

  • Investigated recovery and stability of nitrite and nitrate in whole blood and plasma.
  • Analyzed the relationship between plasma nitrite and nitrate concentrations.
  • Examined potential sources of artifacts in plasma sample assays.

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Main Results:

  • Near-quantitative (87%) and reproducible recovery of nitrite and nitrate from plasma.
  • Nitrite and nitrate are stable in frozen plasma for over 1 year.
  • Nitrite in whole blood is rapidly oxidized (>95% in 1 hour) to nitrate.
  • Plasma nitrite and nitrate concentrations were not correlated in healthy individuals.
  • Plasma samples require deproteinization and background controls to prevent artifactually high readings.

Conclusions:

  • Plasma nitrite determination alone is not meaningful due to rapid oxidation in whole blood.
  • Accurate measurement of plasma nitrite and nitrate requires specific sample preparation techniques, including deproteinization and background controls.
  • Established reference ranges for plasma nitrite (1.3-13 µmol/L) and nitrate (4.0-45.3 µmol/L) in healthy subjects.