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

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Hoang, Thi Phuong Thuy | Barthélemy, Morgane | Lami, Raphaël | Stien, Didier | Eparvier, Véronique | Touboul, David

Edité par CCSD ; Springer Verlag -

International audience. In recent years, supercritical-fluid chromatography (SFC) using CO 2 as the mobile phase is expanding in the research laboratory and industry since it is considered as a green analytical method. This technic offers numerous advantages such as good separation and sensitive detection, short analysis times and stability for analytes. In this study, a method for quantification of lipids, called N-acyl homoserine lactones (AHLs), signal molecules responsible of cell-to-cell communication initially discovered in the kingdom of bacteria, by SFC coupled to high-resolution mass spectrometry (SFC-HRMS) was developed. Optimization of SFC condition together with MS ionization settings to reach the best separation and sensitivity were investigated. The established optimal analysis conditions allow quantification of up to 30 AHLs in a single run within 16 min with excellent linearity (R 2 >0.998) as well as sensitivity (pg level). This method was then applied to study AHLs production of 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 conducted based on SFC-HRMS/MS data. This led to additionally identify four unknown AHLs annotated as N-3-hydroxy-dodecanoylol homoserine lactone, N-3-hydroxy-dodecadienoyl homoserine lactone (3-OH-C12:2-HSL) and two N-3-oxo-dodecenoyl homoserine lactone (3-oxo-C12:1-HSLs).

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