Skip to main content
Log in

Analysis of thiols by HPLC after fluorescent prelabelling with 4-(6-methylnaphthalen-2-yl)-4-oxobuten-2-oic acid

  • Originals
  • Published:
Chromatographia Aims and scope Submit manuscript

Summary

The use of 4-(6-methylnaphthalen-2-yl)-4-oxobuten-2-oic acid as a fluorogenic reagent in pre-column derivatization for the high-performance liquid chromatography (HPLC) of biologically important thiols (L-cysteine, glutathione, N-acetylcysteine, homocysteine and mercaptopropionylglycine) was investigated. The aroylacrylic acid reacts selectively and rapidly (15 min. at room temperature) with the thiol compounds to give stable fluorescent adducts which can be separated by reversedphase HPLC and detected fluorometrically (λex 300nm); λem 445nm). The experimental conditions for the thiol derivatization and chromatographic separation are discussed. Applications to the determination of N-acetylcysteine, mercaptopropionylglycine and cysteine are described.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. D. Dupuy, S. Szabo, J. Liq. Chromatogr.10, 107 (1987).

    Google Scholar 

  2. J. P. Richie, C. A. Lang, Anal. Biochem.163, 9 (1987).

    Google Scholar 

  3. O. H. Drummer, N. Christophidis, J. D. Horowitz, W. J. Lewis, J. Chromatogr.374, 251 (1986).

    Google Scholar 

  4. D. Perret, S. R. Rudge, J. Pharm. Biomed. Anal.3, 3 (1985). and references cited therein.

    Google Scholar 

  5. K. Hayashi, M. Myamoto, Y. Sekine, J. Chromatogr.,338, 161 (1985).

    Google Scholar 

  6. V. Cavrini, R. Gatti, A. M. DiPietra, M. A. Raggi, Chromatographia,23 680 (1987).

    Google Scholar 

  7. P. A. Lewis, A. J. Woodward, J. Maddock, J. Pharm. Sci.,73, 996 (1984).

    Google Scholar 

  8. K. Imai, T. Toyóoka, H. Miyano, Analyst,109, 1365 (1984). and references cited therein.

    Google Scholar 

  9. K. Nakashima, C. Umekawa, H. Yoshida, S. Nakatsuji, S. Akiyama, J. Chromatogr.,414, 11 (1987).

    Google Scholar 

  10. I. A. Cotgreave, P. Moldeus, J. Biochem. Biophys. Methods,13, 231 (1986).

    Google Scholar 

  11. B. Kagedal, M. Kallberg, J. Chromatogr.308, 75 (1984).

    Google Scholar 

  12. T. Toyóoka, K. Imai, Anal. Chem.56, 2461 (1984), and references cited therein.

    Google Scholar 

  13. W. Baeyens, G. Van der Weken, P. De Moerloose, Chromatographia,23, 717 (1987).

    Google Scholar 

  14. W. Baeyens, G. Van Der Weken, B. Linling, P. De Moerloose, Anal. Lett.21 (5), 741 (1988).

    Google Scholar 

  15. R. M. Maiorino, G. L. Waber, H. Aposhian, J. Chromatogr.,374, 297 (1986).

    Google Scholar 

  16. M. Johansson, D. Westerlund, J. Chromatogr.385, 343 (1987).

    Google Scholar 

  17. K. Shimada, M. Tanaka, T. Nambara, Y. Imai, K. Abe, K. Yoshinaga, J. Chromatogr.227, 445 (1982).

    Google Scholar 

  18. Y. Imai, S. Ito, K. Fujita, J. Chromatogr.420, 404 (1987).

    Google Scholar 

  19. A. J. J. Debets, R. Van De Straat, W. H. Voogt, H. Vos, N. P. E. Vermeulen, R. W. Frei, J. Pharm. Biomed. Anal.,6, 329 (1988).

    Google Scholar 

  20. T. Toyoóka, K. Imai, Y. Kawahara, J. Pharm. Biomed. Anal.2, 473 (1984).

    Google Scholar 

  21. F. K. Kirchner, J. H. Bailay, C. J. Cavallito, J. Am. Chem. Soc.,71, 1210 (1949).

    Google Scholar 

  22. A. Dal Pozzo, M. Acquasaliente, G. Donzelli, P. De Maria, M. L. Nicoli, J. Med. Chem.30, 1674 (1987).

    Google Scholar 

  23. V. Cavrini, R. Gatti, P. Roveri, M. R. Cesaroni, Analyst,113, 1447 (1988).

    Google Scholar 

  24. S. M. Makar, K. F. Bassilios, A. Y. Salem, J. Chem. Soc., 2437 (1958).

  25. J. D. Reinheimer, S. Taylor, J. Org. Chem.,19, 802 (1954).

    Google Scholar 

  26. R. H. Costilow, in “Manual of Methods for general Bacteriology”,P. Gerhardt, Ed., American Society for Microbiology, Washington, DC 20006, 1981, p. 74.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Presented in part at “Bononiachem”, XVI Congresso Nazionale di Chimica, Bologna, October 9–14, 1988.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cavrini, V., Gatti, R., Roveri, P. et al. Analysis of thiols by HPLC after fluorescent prelabelling with 4-(6-methylnaphthalen-2-yl)-4-oxobuten-2-oic acid. Chromatographia 27, 185–190 (1989). https://doi.org/10.1007/BF02260443

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02260443

Key Words

Navigation