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 journalArticle

1958 Citations (Scopus)

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 - 2001
Externally publishedYes

Fingerprint

Nitrites
Nitrates
Assays
S-Nitrosothiols
Nitrate Reductase
Vanadium
Fluids
Cell culture
Culture Media
Arginine
Nitric Oxide
Cell Culture Techniques
Derivatives
Detectors
Plasmas
Equipment and Supplies
Oxidation
Serum

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. / Miranda, Katrina M; Espey, Michael G.; Wink, David A.

In: Nitric Oxide - Biology and Chemistry, Vol. 5, No. 1, 2001, p. 62-71.

Research output: Contribution to journalArticle

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