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Gas chromatography-mass spectrometry determination of [15N]ammonia enrichment in blood and urine
Authors:I Nissim  M Yudkoff  W Yang  T Terwilliger  S Segal
Affiliation:1. Division of Biochemical Development and Molecular Diseases, Children''s Hospital of Philadelphia, Philadelphia, Pennsylvania 19104 USA;2. Department of Pediatrics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 USA;3. Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104 USA
Abstract:A rapid gas chromatography-mass spectrometry method for [15N]ammonia analysis is deseribed which is based on the formation of [15N]glutamic acid from ammonia and analysis of isotopic abundance in the N-trifluoroacetyl-n-butylester glutamate derivative. Mean recovery of [15N]ammonia added to either plasma or urine was greater than 99% with a relative standard deviation of less than 10%. The method can be applied to the determination of extremely low levels of ammonia through an isotope dilution technique. The [15N]ammonia abundance of blood and urine was determined in an adult following on oral dose (500 mg) of 15NH4Cl. A peak isotopic abundance of 13 atoms% excess was reached by 30 min. Urinary excretion of [15N]ammonia during the first 4 h after administration of the isotope amounted to 4.1% of the isotope administered.
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