Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 412, Issue 10, Pages 2261-2276Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-019-02265-4
Keywords
Supercritical-fluid chromatography; N-Acyl homoserine lactone; Quorum sensing; Quantification; Molecular networking
Funding
- Agence Nationale de la Recherche [ANR-16-CE29-0002-01 CAP-SFC-MS, ANR-15-CE21-0016]
- Investissement d'Avenir grant [ANR-10-LABX-25-01]
- Swiss National Science Foundation (SNF) [310030E-164289]
- Region Ile-de-France (DIM Analytics)
- Swiss National Science Foundation (SNF) [310030E-164289] Funding Source: Swiss National Science Foundation (SNF)
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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 (R-2 > 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).
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