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Amino Acid Analysis

Precolumn Derivatization Methods

  • Protocol
Protein Sequencing Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 64))

Abstract

The method described in this chapter is based on derivatization of amino acids, produced by hydrolysis of peptides or proteins, with phenylisothiocyanate. This forms phenylthiocarbamyl amino acids, which are then separated by reverse-phase high performance liquid chromatography and quantified from their UV-absorbance at 254 nm.

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References

  1. Spackman, D. H., Stein, W. H., and Moore, S. (1958) Automatic recording apparatus for use in the chromatography of amino acids. Anal. Chem. 30, 1190–1206.

    Article  CAS  Google Scholar 

  2. Hsu, K.-T. and Currie, B. L. (1978) High-performance liquid chromatography of Dns-amino acids and application to peptide hydrolysates. J. Chromatogr. 166, 555–561.

    Article  CAS  Google Scholar 

  3. Lindroth, P. and Mopper, K. (1979) High performance liquid chromatographic determination of subpicomole amounts of amino acids by precolumn fluorescence derivatization with o-phthaldialdehyde. Anal. Chem. 51, 1667–1674.

    Article  CAS  Google Scholar 

  4. Knoop, D. R., Morgan, E. T., Tarr, G. E., and Coon, M. J. (1982) Purification and characterization of a unique isoenzyme of cytochrome P-450 from liver microsomes of ethanol-treated rabbits. J. Biol. Chem. 257, 8472–8480.

    Google Scholar 

  5. Hendrikson, R. L. and Meridith, S. C. (1984) Amino acid analysis by reverse-phase high-performance liquid chromatography: precolumn derivatization with phenylisothiocyanate. Anal. Biochem. 136, 65–74.

    Article  Google Scholar 

  6. Bidlingmeyer, B. A., Cohen, S. A., and Tarvin, T. L. (1984) Rapid analysis of amino acids using pre-column derivatization. J. Chromatogr. 336, 93–104.

    Article  CAS  Google Scholar 

  7. Cohen, S. A. and Strydom, D. J. (1988) Amino acid analysis utilizing phenylisothiocyanate derivatives. Anal. Biochem. 174, l–16.

    Article  Google Scholar 

  8. Strydom, D. J. and Cohen, S. A. (1994) Comparison of amino acid analyses by phenylisothiocyanate and 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate precolumn derivatization. Anal. Biochem. 222, 19–28.

    Article  CAS  Google Scholar 

  9. Hendrikson, R. L., Mora, R., and Maraganore, J. M. (1987) A practical guide to the general application of PTC-amino acid analysis, in Proteins Structure and Function (L’Italien, J. J., ed.), Plenum, New York, pp. 187–195.

    Google Scholar 

  10. Tous, G. I., Fausnaugh, J. L., Akinyosoye, O. Lackland, H., Winter-Cash, P., Vitorica, F. J., and Stein, S. (1989) Amino acid analysis on polyvinylidene difluoride membranes. Anal. Biochem. 179, 50–55.

    Article  CAS  Google Scholar 

  11. Nakagawa, S. and Fukuda, T. (1989) Direct amino acid analysis of proteins electroblotted onto polyvinylidene difluoride membrane from sodium dodecyl sulfate-polyacrylamide gel. Anal. Biochem. 181, 75–78.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Division of Biochemistry, School of Biology and Biochemistry, Medical Biology Centre, Queen’s University of Belfast, Northern Ireland

    G. Brent Irvine

Authors
  1. G. Brent Irvine
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Editors and Affiliations

  1. Celltech Therapeutics, Slough, UK

    Bryan John Smith

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© 1997 Humana Press Inc.

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Irvine, G.B. (1997). Amino Acid Analysis. In: Smith, B.J. (eds) Protein Sequencing Protocols. Methods in Molecular Biology™, vol 64. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-353-8:131

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  • DOI: https://doi.org/10.1385/0-89603-353-8:131

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-353-5

  • Online ISBN: 978-1-59259-550-1

  • eBook Packages: Springer Protocols

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