Spectrophotometric Determination of Serum Nitrite and Nitrate by Copper–Cadmium Alloy

• Published in: Analytical biochemistry Vol. 306; no. 1; pp. 79 - 82
• Main Authors: Sastry, K.V.H., Moudgal, R.P., Mohan, J., Tyagi, J.S., Rao, G.S.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2002
• Subjects: Alloys - chemistry; Animals; Buffaloes; Cadmium - chemistry; Cattle; Chickens; Copper - chemistry; copper–cadmium reduction; Coturnix; Ethylenediamines; Free Radical Scavengers; Goats; Griess reaction; Humans; Nitrates - blood; nitric oxide; Nitric Oxide - chemistry; nitrite and nitrate; Nitrites - blood; Oxidation-Reduction; Rats; Sensitivity and Specificity; Sheep; Spectrophotometry, Ultraviolet - methods; Sulfanilamides; Time Factors; nitric oxide; copper–cadmium reduction; Griess reaction; nitrite and nitrate
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1006/abio.2002.5676

A macro and micro assay for the spectrophotometric determination of serum nitrite and nitrate was developed. Nitrite/nitrate in biological samples can be estimated in a single step by this method. The principle of the assay is the reduction of nitrate by copper–cadmium alloy, followed by color development with Griess reagent (sulfanilamide and N-naphthylethylenediamine) in acidic medium. This assay is sensitive to 1 μM nitrate and is suitable for different biological fluids, including sera with a high lipid concentration. The copper–cadmium alloy used in the present method is easy to prepare and can completely reduce nitrate to nitrite in an hour. The present method provides a simple, cost-effective assay for the estimation of stable oxidation products of nitric oxide in biological samples.