Determination of Nitrate and Nitrite Ions in Human Plasma by Ion Exchange-High Performance Liquid Chromatography

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Abstract
Acetonitrile precipitation of plasma samples followed by injection of supernatant onto a reverse phase precolumn coupled to an anion exchange column allowed ultraviolet detection (214 nm) of eluting nitrate and nitrite ions. Sensitivity in plasma is about 0.01 mM for both ions and linearity is excellent from 0.02 to 1.0 mM. Nitrite accuracy assessed by diazotization coupling was good. Reproducibility studies demonstrated withinrun coefficients of variation of < 4%. Interferences were few. Random endogenous serum nitrate concentrations (0.03–0.12 mM) were determined. Serum nitrite and nitrate concentrations were measured in a patient following an overdosage of isobutyl nitrite. The method is applicable for nitrite/nitrate studies in plasma at these concentrations.