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Nitric Oxide. 2001 Feb;5(1):62-71.

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

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Radiation Biology Branch, National Cancer Institute, National Institutes of Health, Building 10, Room B3-B69, Bethesda, Maryland 20892, USA.


Numerous methods are available for measurement of nitrate (NO(-)(3)). 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 microM NO(-)(3) 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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