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Elimination of matrix-based interferences to a fluorescent nitrite/nitrate assay by a simple filtration procedure
D W Bedwell1, V R Rivera, G A Merrill
1U.S. Army Institute of Surgical Research, 3600 Rawley E. Chambers Avenue, Fort Sam Houston, Texas 78234, USA.
Abstract:
Pathophysiological levels of oxygen radical metabolites have been studied as indicators of trauma caused by burn insult. The 2, 3-diaminonaphthalene assay is routinely used in the determination of nitrite/nitrate levels in biological fluids and cellular extracts as one indicator of nitric oxide activity. Several laboratories, including ours, have noted matrix-based interferences resulting in decreased assay sensitivity during nitrite/nitrate analysis. We evaluated filtration using Millipore Ultrafree-MC 10,000 NMWL filters for the ability to eliminate matrix-based interferences from human serum and tissue culture medium, thereby restoring assay sensitivity.