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

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

doi:10.1006/niox.2000.0319

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 journal › Article › peer-review

3235 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 language	English (US)
Pages (from-to)	62-71
Number of pages	10
Journal	Nitric Oxide - Biology and Chemistry
Volume	5
Issue number	1
DOIs	https://doi.org/10.1006/niox.2000.0319
State	Published - 2001
Externally published	Yes

Keywords

Griess assay
Nitrate
Nitric oxide
Nitrite
Vanadium(III) reduction

ASJC Scopus subject areas

Biochemistry
Physiology
Clinical Biochemistry
Cancer Research

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10.1006/niox.2000.0319

Cite this

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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.",

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AU - Espey, Michael G.

AU - Wink, David A.

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N2 - 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.

AB - 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.

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

Abstract

Keywords

ASJC Scopus subject areas

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