A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite

Katrina M. Miranda, Michael G. Espey, David A. Wink

Research output: Contribution to journalArticlepeer-review

2200 Scopus citations

Abstract

Numerous methods are available for measurement of nitrate (NO3-). However, these assays can either be time consuming or require specialized equipment (e.g., nitrate reductase, chemiluminescent detector). We have developed a method for simultaneous evaluation of nitrate and nitrite concentrations in a microtiter plate format. The principle of this assay is reduction of nitrate by vanadium(III) combined with detection by the acidic Griess reaction. This assay is sensitive to 0.5 μM NO3- and is useful in a variety of fluids including cell culture media, serum, and plasma. S-Nitrosothiols and L-arginine derivatives were found to be potential interfering agents. However, these compounds are generally minor constituents of biological fluids relative to the concentration of nitrate/nitrite. This report introduces a new, convenient assay for the stable oxidation products of nitrogen oxide chemistry in biological samples.

Original languageEnglish (US)
Pages (from-to)62-71
Number of pages10
JournalNitric Oxide - Biology and Chemistry
Volume5
Issue number1
DOIs
StatePublished - Jan 1 2001
Externally publishedYes

Keywords

  • Griess assay
  • Nitrate
  • Nitric oxide
  • Nitrite
  • Vanadium(III) reduction

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Clinical Biochemistry
  • Cancer Research

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