Abstract
The fate of nitrate in various tissues of the mouse after administration of Na15NO2 or K15NO3 is described. The method for determining NO3-15N is based on the formation of 4-15NO2-2-sec-butylphenol by the reaction of nitrate and 2-sec-butylphenol in aqueous (5:7) sulfuric acid at room temperature for 15min and subsequent trimethylsilylation to form the trimethylsilyl ester of 4-15NO2-2-sec-butylphenol, which is then determined by gas chromatography mass spectrometry with selected ion monitoring (m/e 239). Amounts of 0.3-5.0μg of NO3-15N can be determined. The detection limit of NO3-15N for the whole procedure was 0.08μg/ml. This is a specific method for NO3-15N. We used this method to carry out a metabolic fate study of nitrate in various tissues of mouse following a single oral dose of 100μg or 500μg of NO2-15N or NO3-15N. The NO3-15N concentrations in brain, lung, spleen, heart, the kidney and liver were followed for 180min after oral administration. In the case of Na15NO2 administration, the amounts of NO3-15N detected in various tissues derived from K15NO3 reached the maximum values at 30min after administration, then gradually decreased. In the case of K15NO3 administration, the amounts of NO3-15N detected in various tissues reached the maximum values at 45min after administration, then gradually decreased.
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