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Nitrate analysis by capillary gas chromatography.

M J Dunphy1, D D Goble, D J Smith

  • 1Department of Chemistry, University of Akron, Ohio 44325.

Analytical Biochemistry
|February 1, 1990
PubMed
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A new microanalytical gas chromatographic method accurately quantifies nitrate in rat urine. This sensitive technique offers reliable nitrate analysis for biological samples, aiding toxicological and metabolic research.

Area of Science:

  • Analytical Chemistry
  • Environmental Science
  • Toxicology

Background:

  • Nitrate analysis in biological samples is crucial for understanding physiological and toxicological processes.
  • Existing methods may lack sensitivity or be prone to interference.
  • A need exists for a precise and reliable method for urinary nitrate determination.

Purpose of the Study:

  • To develop and validate a microanalytical gas chromatographic (GC) method for quantifying nitrate in rat urine.
  • To establish the method's sensitivity, linearity, precision, and accuracy.
  • To assess the method's applicability in biological studies.

Main Methods:

  • Nitrate in rat urine was converted to nitromesitylene.
  • Quantification was performed using 3,4-dimethylnitrobenzene as an internal standard via GC with a nitrogen-phosphorus thermionic detector.

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  • Sample cleanup involved silver acetate to remove chloride interference.

    • Main Results:

    • The method demonstrated linearity from 1.0 to 1000 microM nitrate with a detection limit of 0.5 microM.
    • High precision was observed with intra-day coefficients of variation (CV) ranging from 2.2% to 6.5% and inter-day CVs from 2.0% to 6.1%.
    • Average recovery rates were 77% and 80%, with results comparable to colorimetric assays.

    Conclusions:

    • The developed GC method is a sensitive, precise, and accurate technique for microanalytical determination of nitrate in rat urine.
    • The method is robust, with stable samples for up to 60 days and minimal interference.
    • This validated method is suitable for measuring urinary nitrate levels in toxicological and metabolic research involving rodents.