Liquid Chromatography/Mass Spectrometry for the Detection and Quantification of N-Acyl-l-Homoserine Lactones and 4-Hydroxy-2-Alkylquinolines

  • François Lépine
  • Eric DézielEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 692)


High-performance liquid chromatography (HPLC) coupled in-line with mass spectrometry (MS) permits rapid and specific identification and quantification of N-acyl-l-homoserine lactones (AHLs) and 4-hydroxy-2-alkylquinolines (HAQs). We are presenting here methods for the analysis of these molecules directly from biological samples using LC/MS.

Key words

Bacteria Quorum sensing Acyl homoserine lactone 4-Hydroxy-2-alkylquinoline Analysis Quantification Mass spectrometry Liquid chromatography 



The important contribution of Sylvain Milot to the development of these methods is gratefully acknowledged. We also thank Émilie Gauthier for the help in the development of the AHLs quantification methods.


  1. 1.
    Pearson, J. P., Gray, K. M., Passador, L., Tucker, K. D., Eberhard, A., Iglewski, B. H., and Greenberg, E. P. (1994) Structure of the autoinducer required for expression of Pseudomonas aeruginosa virulence genes, Proc Natl Acad Sci U S A 91, 197–201.PubMedCrossRefGoogle Scholar
  2. 2.
    Pearson, J. P., Passador, L., Iglewski, B. H., and Greenberg, E. P. (1995) A second N-acylhomoserine lactone signal produced by Pseudomonas aeruginosa, Proc Natl Acad Sci U S A 92, 1490–1494.PubMedCrossRefGoogle Scholar
  3. 3.
    Déziel, E., Lépine, F., Milot, S., He, J., Mindrinos, M. N., Tompkins, R. G., and Rahme, L. G. (2004) Analysis of Pseudomonas aeruginosa 4-hydroxy-2- alkylquinolines (HAQs) reveals a role for 4-hydroxy-2-heptylquinoline in cell-to-cell communication, Proc Natl Acad Sci U S A 101, 1339–1344.PubMedCrossRefGoogle Scholar
  4. 4.
    Diggle, S. P., Lumjiaktase, P., Dipilato, F., Winzer, K., Kunakorn, M., Barrett, D. A., Chhabra, S. R., Camara, M., and Williams, P. (2006) Functional genetic analysis reveals a 2-alkyl-4-quinolone signaling system in the human pathogen Burkholderia pseudomallei and related bacteria, Chem Biol 13, 701–710.PubMedCrossRefGoogle Scholar
  5. 5.
    Pesci, E. C., Milbank, J. B. J., Pearson, J. P., McKnight, S., Kende, A. S., Greenberg, E. P., and Iglewski, B. H. (1999) Quinolone signaling in the cell-to-cell communication system of Pseudomonas aeruginosa, Proc Natl Acad Sci U S A 96, 11229–11234.PubMedCrossRefGoogle Scholar
  6. 6.
    Vial, L., Lépine, F., Milot, S., Groleau, M. C., Dekimpe, V., Woods, D. E., and Déziel, E. (2008) Burkholderia pseudomallei, B. thailandensis, and B. ambifaria produce 4-hydroxy-2-alkylquinoline analogues with a methyl group at the 3 position that is required for quorum-sensing regulation, J Bacteriol 190, 5339–5352.PubMedCrossRefGoogle Scholar
  7. 7.
    Rahme, L. G., Stevens, E. J., Wolfort, S. F., Shao, J., Tompkins, R. G., and Ausubel, F. M. (1995) Common virulence factors for bacterial pathogenicity in plants and animals, Science 268, 1899–1902.PubMedCrossRefGoogle Scholar
  8. 8.
    Lépine, F., Déziel, E., Milot, S., and Rahme, L. G. (2003) A stable isotope dilution assay for the quantification of the Pseudomonas quinolone signal in Pseudomonas aeruginosa cultures, Biochim Biophys Acta 1622, 36–41.PubMedCrossRefGoogle Scholar
  9. 9.
    Xiao, G., Déziel, E., He, J., Lépine, F., Lesic, B., Castonguay, M. H., Milot, S., Tampakaki, A. P., Stachel, S. E., and Rahme, L. G. (2006) MvfR, a key Pseudomonas aeruginosa pathogenicity LTTR-class regulatory protein, has dual ligands, Mol Microbiol 62, 1689–1699.PubMedCrossRefGoogle Scholar
  10. 10.
    Lépine, F., Milot, S., Déziel, E., He, J., and Rahme, L. G. (2004) Electrospray/mass spectrometric identification and analysis of 4-hydroxy-2-alkylquinolines (HAQs) produced by Pseudomonas aeruginosa, J Am Soc Mass Spectrom 15, 862–869.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.INRS-Institut Armand-FrappierLavalCanada

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