Abstract
N-carbamylglutamate (NCG), a synthetic analogue of N-acetylglutamate, is an activator of blood ammonia conversion and endogenous arginine synthesis. Here, we established an accurate quantitative determination of NCG in feeds, animal tissues, and body fluids using the high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The sample pretreatment procedures included extraction with 0.5% of formic acid in water/methanol (80/20, v/v), and purification using an anionic solid phase extraction cartridge. Satisfactory separation of NCG was achieved in 20 min with the application of an Atlantis T3 column, and a confirmative detection of NCG was ensured by multiple reaction monitoring of positive ions. NCG spiked in feeds, tissues, and body fluids were evaluated in regard to linearity, sensitivity, recovery, and repeatability. Recoveries for different sample matrices were in the range of 88.12% to 110.21% with relative standard deviations (RSDs) less than 8.8%. Limits of quantification were within the range of 0.012 to 0.073 mg kg−1 and 0.047 to 0.077 μg mL−1 for solid and liquid samples, respectively. This study will provide a solid foundation for the evaluation of availability and metabolic mechanism of NCG in animals.
Highlights
N-carbamylglutamate (NCG) is a synthetic analogue of N-acetylglutamate (NAG) (Figure 1), which is the allosteric stimulator of carbamyl phosphate synthase-1 (CPS1)
CPS1 is a key enzyme functioning in the urea cycle and endogenous arginine synthesis pathway [1]
Due to its good stability and safety [2,3], NCG was initially used for the clinical treating of hyperammonemia caused by the NAG synthetase deficiency [4,5], propionic aciduria and methylmalonic aciduria [6,7], and maple syrup urine disease [8]
Summary
N-carbamylglutamate (NCG) is a synthetic analogue of N-acetylglutamate (NAG) (Figure 1), which is the allosteric stimulator of carbamyl phosphate synthase-1 (CPS1). Like NAG, NCG can activate CPS1 and lead the conversion of blood ammonia into the mitochondrial carbamoyl-phosphate, further stimulating the endogenous synthesis of arginine. NCG is proven to be a novel, effective and low-cost substitute feed additive for arginine. NCG on growth promotion performance enhancement have been found in other species of animals including the rat [12], and reproductive performance enhancement have been found in other species of animals cattle [13], the sheep yellow-feather broiler including rat [14],. Tried to the determine the NCG using will result will in low responsivity and sensitivity. NCG intissues, the feed, tissues, andexcellent body fluids with quantitatively determine. NCG indetermine the feed, animal andanimal body fluids with accuracy excellent accuracy sensitivity. In the feeds and animal as well as the accuracyas and repeatability for NCG in repeatability the feeds andfor animal products
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