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The Determination of Polyamines and Amino Acids by a Fluorescamine-HPLC Method

  • Karl J. Hunter
  • Alan H. Fairlamb
Part of the Methods in Molecular Biology™ book series (MIMB, volume 79)

Abstract

The derivatization of polyamines and amino acids with fluorescamine is a useful tool for the detection and quantitation of these compounds. High-pressure liquid chromatography (HPLC) separation coupled to postcolumn derivatization allows for the detection of nanomole amounts of material and is simple to use. Because the compounds of interest are derivatized after separation by HPLC, their biological functionality is not destroyed prior to analysis, allowing the mixture to be separated for other uses. This method also has the advantage of being able to detect small peptides and conjugated polyamines (e.g., glutathione and trypanothione) and with the use of acid hydrolysis, allows for their composition to be analyzed.

Keywords

Amino Acid Mixture Lithium Borate Pressure Relief Valve Back Pressure Regulator Camphorsulfonic Acid 
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.

References

  1. 1.
    Westhall, F. and Hesser, H. (1974) Fifteen minute aced hydrolysis of peptides. Anal. Biochem. 61, 610–613.CrossRefGoogle Scholar
  2. 2.
    Weigele, M., Debernado, S., and Leimgruber, W. (1973) Fluorometric assay of secondary amino acids. Biochem. Biophys. Res. Commun. 50, 352–356.PubMedCrossRefGoogle Scholar
  3. 3.
    Berger, B. J. and Fairlamb, A. H. (1988) High performance liquid chromatographic method for the separation and quantitative estimation of anti-parasitic melaminophenyl arsenical compounds. Trans. Roy. Soc. Trop. Med. Hyg. 88, 357–359.CrossRefGoogle Scholar
  4. 4.
    Berger, B. J. and Fairlamb, A. H. (1994) Properties of melarsamine hydrochloride (Cymelarsan) in aqueous solution. Antimicrob. Agents Chemother. 38, 1298–1302.PubMedGoogle Scholar
  5. 5.
    Fairlamb, A. H., Henderson, G. B., and Cerami A. (1989) Trypanothione is the primary target for arsenical drugs against African trypanosomes. Proc. Natl. Acad. Sci. USA. 86, 2607–2611.PubMedCrossRefGoogle Scholar
  6. 6.
    Fairlamb, A. H., Smith, K., and Hunter, K. J. (1992) The interaction of arsenical drugs with dihydrolipoamide and dihydrolipoamide dehydrogenase from arsenical resistant and sensitive strains of Trypanosoma brucei brucei Molec. Biochem. Parasitol 53, 223–232.CrossRefGoogle Scholar
  7. 7.
    Cunningham, M. L. and Fairlamb, A. H. (1995) Trypanothione reductase from Leishmania donovani—purification, characterisation and inhibition by trivalent antimonials. Eur. J. Biochem. 230, 460–468.PubMedCrossRefGoogle Scholar
  8. 8.
    Hunter, K. J. and Fairlamb, A. H. (1990) Separation and quantitation of the polyamine biosynthesis inhibitor DL-α-difluoromethylarginine and other guanidine-containing compounds by high-performance liquid chromatography. Anal. Biochem. 190, 281–285.PubMedCrossRefGoogle Scholar
  9. 9.
    Fairlamb, A. H., Blackburn, P., Ulrich, P., Chait, B. T., and Cerami, A. (1985) Trypanothione a novel bis(glutathionyl)spermidine cofactor for glutathione reductase in trypanosomatids. Science 227, 1485–1487.PubMedCrossRefGoogle Scholar
  10. 10.
    Hunter, K. J., Le Quesne, S. A., and Fairlamb, A. H. (1994) Identification and biosynthesis of N1,N9-bis(glutathionyl)aminopropylcadaverine (homotrypanothione) in Trypanosoma cruzi. Eur. J. Biochem. 226, 1019–1027.PubMedCrossRefGoogle Scholar
  11. 11.
    Fairlamb, A. H., Henderson, G. B., Bacchi, C. J., and Cerami, A. (1987) In vivo effects of difluoromethylornithine on trypanothione and polyamine levels in bloodstream forms of Trypanosoma brucei. Mol. Biochem. Parasitol. 24, 185–191.PubMedCrossRefGoogle Scholar
  12. 12.
    Shim, H. and Fairlamb, A. H. (1988) Levels of polyamines, glutathione and glutathione-spermidine conjugates during growth of the insect trypanosomatid Crithidia fasciculata. J. Gen. Microbiol. 134, 807–817.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 1998

Authors and Affiliations

  • Karl J. Hunter
    • 1
  • Alan H. Fairlamb
    • 2
  1. 1.Biotransformation Unit, Unilever ResearchSharnbrookUK
  2. 2.Department of Medical ParasitologyLondon School of Hygiene and Tropical MedicineLondonUK

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