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Nitro-L-arginine is used as an inhibitor of nitric oxide synthase in many biological Systems. Nitric oxide (NO) is unstable and degrades to nitrite NO(2)- and nitrate NO(3)-. The colorimetric reaction of N0(2)- with Griess reagent is commonly used to measure NO(2)-. NO(3)- may be measured as NO(2)- following reduction by cadmium or cadmium/copper. We found that bradykinin increased the formation of NO(2)- by bovine coronary endothelial cells. Nitro-L-arginine further increased the formation of NO(2)-. This increase is due to the interference of nitro-L-arginine in determination of NO(3)- by the cadmium reduction to NO(2)- and Griess reagent reaction. Incubation of nitro-L-arginine with cadmium or cadmium/copper produced a product that reacts with Griess reagent to form a compound that has an absorption spectrum identical to the product formed by NO(2)- and Griess reagent. Caution must be exercised when using the NO(2)-/NO(3)- measurement by the Griess reaction to assess inhibition of nitric oxide synthase by nitro-L-arginine.

Nitro- L -Arginine Interferes with the Cadmium Reduction of Nitrate/ Griess Reaction Method of Measuring Nitric Oxide Production