Elimination of matrix-based interferences to a fluorescent nitrite/nitrate assay by a simple filtration procedure.
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.
Bedwell
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DW
U.S. Army Institute of Surgical Research, 3600 Rawley E. Chambers Avenue, Fort Sam Houston, Texas 78234, USA.
Rivera
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VR
Merrill
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GA
Pusateri
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AE
D015081:2-Naphthylamine / Q000031:analogs & derivatives / Q000032:analysis; D000818:Animals; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D005374:Filtration / Q000379:methods*; D006801:Humans; D008851:Micropore Filters; D009566:Nitrates / Q000032:analysis* / Q000097:blood; D009573:Nitrites / Q000032:analysis* / Q000097:blood; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000494:pharmacology; D013050:Spectrometry, Fluorescence / Q000379:methods; D014462:Ultrafiltration / Q000379:methods*
D009566:Nitrates; D009573:Nitrites; D012709:Serum Albumin; C026373:2,3-diaminonaphthalene; D015081:2-Naphthylamine