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[Assay validation for determining nitrites and nitrates in biological fluids]
Rosa María Muñoz-Fuentes1, Florencia Vargas, Norma A Bobadilla
1Departamento de Gastroenterología, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán.
Summary
This study presents a cost-effective and accessible assay for measuring nitrites and nitrates, crucial indicators of nitric oxide production. The new method offers comparable results to commercial kits, making it a viable alternative for laboratories worldwide.
Area of Science:
- Biochemistry
- Analytical Chemistry
Context:
- Nitric oxide (NO) is a key signaling molecule in physiological and pathophysiological processes.
- Quantifying stable NO metabolites, nitrites (NO2-) and nitrates (NO3-), serves as an indirect measure of NO production.
- Existing commercial kits for NO2- + NO3- quantification are expensive and not universally accessible.
Purpose:
- To develop and validate an economical and accessible assay for determining nitrites and nitrates in biological fluids.
- To provide a cost-effective alternative to commercial kits for NO metabolite analysis.
Summary:
- The study describes an enzymatic method using nitrate reductase followed by Griess reagent for NO2- quantification.
- Assay validation involved known sodium nitrate concentrations and analysis of plasma and ascites samples from cirrhotic patients.
- The developed assay demonstrated reliable detection ranges for nitrates (25-400 µM/L) and nitrites (25-100 µM/L) with high recovery rates (>95%) in spiked plasma samples.
Impact:
- The validated assay provides accurate and reproducible measurements of NO2- + NO3- in plasma and ascites.
- This method is cost-effective, requiring no special equipment, thus enhancing accessibility for laboratories with limited resources.
- The assay's cost-effectiveness is approximately one-fifth of that of commercial kits, promoting wider adoption in clinical and research settings.