Oxidation of ammonia and hydroxylamine to nitrate in the rat and in vitro.
We have demonstrated that ammonia is oxidized to nitrate in the rat. Male Sprague-Dawley rats gavaged with 1000 mumol N-15-ammonium chloride each day for 5 days were found to excrete low, but significant amounts of excess N-15-nitrate in their urines on the five days of treatment and on the five subsequent days. We recovered a total of 0.28 +/- 0.03 mumol excess N-15-nitrate (mean +/- SE) per rat, which indicates that ammonia is converted to nitrate in a yield of approximately 0.0080%. The oxidation of N-15-labeled glycine and L-glutamic acid to N-15-nitrate could not be detected. N-15-Hydroxylamine was oxidized in the rat to N-15-nitrate in a yield of 4.7%, which shows that hydroxylamine is a possible intermediate in the ammonia oxidation process. Injection of rats with Arochlor 1254, an inducer of several isozymes of cytochrome P-450, did not significantly affect the rate of endogeneous nitrate synthesis. Carbon tetrachloride, which causes hepatic lipid peroxidation, produced a small but significant increase in nitrate synthesis. We confirmed the observation that a bacterial endotoxin can greatly stimulate nitrate synthesis, and we showed that concurrent treatment with superoxide dismutase does not modify the effect of the endotoxin. An in vitro chemical model system was used to demonstrate that oxidation of ammonia to nitrate by the hydroxyl radical at physiological pH is chemically feasible. Our results are consistent with the hypothesis that ammonia is oxidized to nitrate in vivo by a non-enzymatic process which involves active oxygen species such as the hydroxyl radical. We estimate that a 215 g rat produces 3.0 mumol of nitrate per day via ammonia oxidation.
D000641:Ammonia / Q000378:metabolism*; D000818:Animals; D002251:Carbon Tetrachloride / Q000494:pharmacology; D004731:Endotoxins / Q000494:pharmacology; D006898:Hydroxylamines / Q000378:metabolism*; D009566:Nitrates / Q000378:metabolism* / Q000652:urine; D010084:Oxidation-Reduction; D010100:Oxygen; D051381:Rats; D013482:Superoxide Dismutase / Q000378:metabolism
D004731:Endotoxins; D006898:Hydroxylamines; D009566:Nitrates; D000641:Ammonia; D002251:Carbon Tetrachloride; D013482:Superoxide Dismutase; D010100:Oxygen