Elsevier

Analytical Biochemistry

Volume 33, Issue 2, February 1970, Pages 263-272
Analytical Biochemistry

Simultaneous determination of citrulline and urea using diacetylmonoxime

https://doi.org/10.1016/0003-2697(70)90296-4Get rights and content

Abstract

A rapid method of simultaneously determining concentrations of citrulline and urea in biological fluids is presented. This method and the accompanying nomograph are based on urea and citrulline absorption peaks at 478 and 490 nm, respectively. Using the diacetylmonoxime method it is possible to determine 0.005 to 0.10 μmole citrulline/ml and 0.025 to 0.07 μmole urea/ml, without dilution. Reactions with other chromogens are discussed.

References (21)

  • R.M. Archibald

    J. Biol. Chem

    (1944)
  • R.M. Archibald et al.

    J. Biol. Chem

    (1945)
  • S. Grisolia
  • J.J. Hagan et al.

    Anal. Biochem

    (1968)
  • D. Hunninghake et al.

    Anal. Biochem

    (1966)
  • D.J. Marsh et al.

    Anal. Biochem

    (1965)
  • S. Ratner
  • P. Righetti et al.

    Anal. Biochem

    (1968)
  • F. Roch-Ramel

    Anal. Biochem

    (1967)
  • P.S. Satoh et al.

    Anal. Biochem

    (1968)
There are more references available in the full text version of this article.

Cited by (59)

  • Impact of suitable control on a uniform interpretation of units for arginase assay

    2021, Biochemistry and Biophysics Reports
    Citation Excerpt :

    The activity of arginase serves as a diagnostic marker which is measured by colorimetric estimation of either arginine (substrate) or ornithine and urea (products) [8]. Preferably, ornithine is estimated either by the method of Chinard [9] or by Archibald [10], and the urea is determined by the direct method of Diacetyl Monooxime [11] or by the indirect methods which convert urea into ammonia and detection of ammonia either by the Nessler's reagent or by the Berthelot's method. Various improvements have been made over time to overcome limitations of the reaction between the component, prolonged boiling, low sensitivity, and use of corrosive materials, but the uniform interpretation of units for arginase assay needs attention.

  • Antioxidant activity of endogenously produced nitric oxide against the zinc oxide nanoparticle-induced oxidative stress in primary hepatocytes of air-breathing catfish, Clarias magur

    2019, Nitric Oxide - Biology and Chemistry
    Citation Excerpt :

    Both sets of reaction mixtures were incubated at 27 °C for 20 min, and the reaction was stopped by adding 1 mL 10% PCA to precipitate out the protein, followed by centrifugation at 10,000×g for 5 min. Citrulline, so formed as the reaction product, was estimated in the supernatant spectrophotometrically at 490 nm in a UV-visible spectrophotometer (Cary 60, Agilent) following the method of Moore and Kauffman [20] and expressed as enzyme activity. The part of the activity, which was inhibited in the second set of the reaction mixture, was taken as iNOS activity.

  • Unique mitochondrial localization of arginase 1 and 2 in hepatocytes of air-breathing walking catfish, Clarias batrachus and their differential expression patterns under hyper-ammonia stress

    2017, Gene
    Citation Excerpt :

    The reaction was stopped by adding 10% perchloric acid at 1:0.5 ratio in the reaction mixture after 20 min of incubation at 27 °C and centrifuged to precipitate out the protein. Urea formed by the reaction was estimated in the supernatant following the method of Moore and Kauffman (1970). One unit of arginase activity was expressed as that amount which catalyzed 1 μmole of urea formed per h at 27 °C.

  • Expression of inducible nitric oxide synthase and nitric oxide production in the mud-dwelled air-breathing singhi catfish, Heteropneustes fossilis under condition of water shortage

    2012, Nitric Oxide - Biology and Chemistry
    Citation Excerpt :

    Both sets of reaction mixtures were incubated at 30 °C for 20 min and the reactions were stopped by adding 1 ml 10% perchloric acid (PCA) to precipitate out the protein, followed by centrifugation at 5,000×g for 10 min. Citrulline, so formed as the reaction product, was estimated in the supernatant spectrophotometrically at 490 nm following the method of Moore and Kauffman [29] and expressed as enzyme activity. The part of activity, which was inhibited by the presence of aminoguanidine (a specific inhibitor of iNOS) from the total activity, was taken as iNOS activity.

  • Influence of environmental ammonia on the production of nitric oxide and expression of inducible nitric oxide synthase in the freshwater air-breathing catfish (Heteropneustes fossilis)

    2012, Aquatic Toxicology
    Citation Excerpt :

    Both sets of reaction mixtures were incubated at 30 °C for 20 min and the reactions were stopped by adding 1 ml 10% perchloric acid (PCA) to precipitate out the protein, followed by centrifugation at 5000 × g for 10 min. Citrulline, so formed as the reaction product, was determined in the supernatant spectrophotometrically at 490 nm following the method of Moore and Kauffman (1970) and expressed as enzyme activity. The part of activity, which was inhibited by the presence of aminoguanidine (a specific inhibitor of iNOS) from the total activity, was taken as iNOS activity.

View all citing articles on Scopus
1

Associate Research Scientist.

View full text