Annotation and quantification of N-acyl homoserine lactones implied in bacterial quorum sensing by supercritical-fluid chromatography coupled with high-resolution mass spectrometry

  • Thi Phuong Thuy Hoang
  • Morgane Barthélemy
  • Raphaël Lami
  • Didier Stien
  • Véronique Eparvier
  • David TouboulEmail author
Research Paper
Part of the following topical collections:
  1. Current Progress in Lipidomics


In recent years, use of supercritical-fluid chromatography (SFC) with CO2 as the mobile phase has been expanding in the research laboratory and industry since it is considered to be a green analytical method. This technique offers numerous advantages, such as good separation and sensitive detection, short analysis times, and stability of analytes. In this study, a method for quantification of N-acyl homoserine lactones (AHLs), signaling molecules responsible for cell-to-cell communication initially discovered in bacteria, by SFC coupled with high-resolution mass spectrometry (HRMS) was developed. The SFC conditions and MS ionization settings were optimized to obtain the best separation and greatest sensitivity. The optimal analysis conditions allowed quantification of up to 30 AHLs in a single run within 16 min with excellent linearity (R2 > 0.998) and sensitivity (picogram level). This method was then applied to study AHL production by one Gram-negative endophytic bacterium, Paraburkholderia sp. BSNB-0670. Nineteen known AHLs were detected, and nine abundant HSLs were quantified. To further investigate the production of uncommon AHLs, a molecular networking approach was applied on the basis of the SFC–HRMS/MS data. This led to additional identification of four unknown AHLs annotated as N-3-hydroxydodecanoylol homoserine lactone, N-3-hydroxydodecadienoyl homoserine lactone, and N-3-oxododecenoyl homoserine lactones (two isomers).


Supercritical-fluid chromatography N-Acyl homoserine lactone Quorum sensing Quantification Molecular networking 



This work was supported by the Agence Nationale de la Recherche (grant ANR-16-CE29-0002-01 CAP-SFC-MS), an Investissement d’Avenir grant managed by Agence Nationale de la Recherche (CEBA, ref ANR-10-LABX-25-01), a joint Agence Nationale de la Recherche and Swiss National Science Foundation (SNF) grant (SECIL, reference ANR-15-CE21-0016 and SNF no. 310030E-164289), and a grant from Région Ile-de-France (DIM Analytics).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Institut de Chimie des Substances Naturelles, CNRS UPR 2301, Univ. Paris‐Sud, Université Paris-SaclayAvenue de la TerrasseGif-sur-YvetteFrance
  2. 2.Sorbonne Université, CNRS, Laboratoire de Biodiversité et Biotechnologies Microbiennes, USR3579, Observatoire Océanologique de Banyuls-sur-MerBanyuls-sur-MerFrance

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