Journal of Inherited Metabolic Disease

, Volume 9, Issue 1, pp 31–38 | Cite as

High-performance liquid chromatographic assay of argininosuccinate: its application in argininosuccinic aciduria and in normal man

  • M. Portolés
  • V. Rubio


Argininosuccinate (ASA) is determined in a few minutes with little manipulation by reversed phase high-performance liquid chromatography (HPLC) usingO-phthaldialdehyde. The two cyclic anhydrides of ASA are not formed during analysis but, if present, can be determined simultaneously. As little as 1 nl urine from a patient with argininosuccinic aciduria was sufficient for analysis; the ASA/creatinine ratio was 50.8 mmol g−1 and daily excretion was 5–7g ASA. We found small amounts of the two anhydrides in the patient's urine and we give factors to estimate, from their peaks, the corresponding amount of ASA. Urine from normal children showed a small acid-labile (at 100°C) peak at the ASA position, which we tentatively assign to genuine ASA. From this peak <2mg ASAday−1 were excreted in our controls. Procedures for collection and storage of samples and the potential of this method for heterozygote detection are discussed.


Public Health Internal Medicine Liquid Chromatography Anhydride Metabolic Disease 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Cusworth, D. C. and Westall, R. Q. Determination of argininosuccinic acid by ion-exchange chromatography.Nature, Lond. 192 (1961) 555–556Google Scholar
  2. Dhondt, J. L., Cartigny, B., Ardouin, P. and Farriaux, J. P. Méthodes d'exploration biochimique des anomalies du cycle de l'urée. In Farriaux, J. P. (ed.)Le Cycle de l'Urée et ses Anomalies, Doin Editeurs, Paris, 1978, pp. 231–263Google Scholar
  3. Hogan, D. L., Kraemer, K. L. and Isenberg, J. I. The use of high-performance liquid chromatography for quantitation of plasma amino acids in man.Anal. Biochem. 127 (1982) 17–24PubMedGoogle Scholar
  4. Nuzum, C. T. and Snodgrass, P. J. Multiple assays of the five urea-cycle enzymes in human liver homogenates. In Grisolía, S., Báguena, R. and Mayor, F. (eds.)The Urea Cycle, John Wiley, New York, 1976, pp. 325–349Google Scholar
  5. Palmer, T., Oberholzer, V. G., Levin, B. and Burgess, E. A. Urinary excretion of arginino-succinic acid.Clin. Chim. Acta 47 (1973) 443–448PubMedGoogle Scholar
  6. Perry, T. L., Wirtz, M. L. K., Kennaway, N. G., Hsia, Y. E., Atienza, F. C. and Uemura, H. S. Amino acid and enzyme studies of brain and other tissues in an infant with argininosuccinic aciduria.Clin. Chim. Acta 105 (1980) 257–267PubMedGoogle Scholar
  7. Plheim, E. and Colombo, J. P. Determination of creatinine in serum, plasma, and urine by a modified Jaffé method without deproteination. In Richterich, R. and Colombo, J. P. (eds.)Clinical Chemistry, John Wiley, New York, 1981, pp. 687–694Google Scholar
  8. Ratner, S. Determination of argininosuccinate in normal blood serum and liver.Analyt. Biochem. 63 (1975) 141–155PubMedGoogle Scholar
  9. Ratner, S. Enzymes of arginine and urea synthesis. In Grisolía, S., Báguena, R. and Mayor, F. (eds)The Urea Cycle, John Wiley, New York, 1976, pp. 181–219Google Scholar
  10. Ratner, S. and Kunkemueller, M. Separation and properties of argininosuccinate and its two anhydrides and their detection in biological materials.Biochemistry 5 (1966) 1821–1832PubMedGoogle Scholar
  11. Shih, V. E. Urea cycle disorders and other congenital hyperammonemic syndromes. In Stanbury, J. B., Wyngaarden, J. B. and Frederickson, D. S. (eds.)The Metabolic Basis of Inherited Disease, 4th Edn., McGraw-Hill, New York, 1978, pp. 362–386Google Scholar
  12. Turnell, D. C. and Cooper, J. D. H. Rapid assay for amino acids in serum or urine by pre-column derivatization and reversed-phase liquid chromatography.Clin. Chem. 28 (1982) 527–531PubMedGoogle Scholar
  13. Van Pilsum, J. F. and Halberg, F. A method for the determination of argininosuccinic acid in human urine.Am. J. Ment. Defic. 67 (1962) 82–89PubMedGoogle Scholar
  14. Walser, M. Urea cycle disorders and other hereditary hyperammonemic syndromes. In Stanbury, J. B., Wyngaarden, J. B., Frederickson, D. S., Goldstein, J. L. and Brown, M. S. (eds.)The Metabolic Basis of Inherited Disease, 5th Edn., McGraw Hill, New York, 1983, pp. 402–438Google Scholar

Copyright information

© SSIEM and MTP Press Limited 1986

Authors and Affiliations

  • M. Portolés
    • 1
  • V. Rubio
    • 1
  1. 1.Instituto de Investigaciones Citológicas de la Caja de Ahorros de ValenciaValenciaSpain

Personalised recommendations