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  • Title: Nitrate analysis in biological fluids by gas chromatography-nitrogen-phosphorous detection.
    Author: Johnson AA, Burleson DG.
    Journal: Anal Biochem; 1996 May 01; 236(2):331-7. PubMed ID: 8660513.
    Abstract:
    Nitrate analysis in body fluids is an important part of the study of nitric oxide metabolism. A sensitive, simple procedure for nitrate analysis was developed by producing nitrobenzene from the nitrate in the samples and benzene. The nitrobenzene produced was measured on a gas chromatograph (GC) equipped with a nitrogen-phosphorous detector. Cl-, which interfered with the derivitization procedure, was removed as silver chloride after precipitation with silver lactate. Rat urine samples were also analyzed after elution through a C18 reverse-phase sample preparation column to remove organic molecules that competed with benzene for the nitrate in the sample. A benzene derivative, 2-nitro-1,3-dimethylbenzene (2-nitro-m-xylene), was chosen as an internal standard for GC analysis. Recovery of nitrate was 67 +/- 2% in serum and 87 +/- 8% in urine. Sensitivity of the procedure was 1 microM. This procedure offers improvements in sensitivity, simplicity, and consistency over previously published procedures and makes possible the measurement of nitrate and determination of isotopic ratios by GC-MS on the same sample.
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