Development and application of a method for the analysis of N-acylhomoserine lactones by solid-phase extraction and ultra high pressure liquid chromatography

Xiaojing Li, Agnes Fekete, Matthias Englmann, Christine Götz, Michael Rothballer, Moritz Frommberger, Katharina Buddrus, Jenoe Fekete, Chunping Cai, Peter Schröder, Anton Hartmann, Guonan Chen, Philippe Schmitt-Kopplin

Research output: Contribution to journalArticlepeer-review

76 Scopus citations

Abstract

A robust method based on solid-phase extraction (SPE) followed by ultra high pressure liquid chromatography (with trade name of Ultra Performance Liquid Chromatography: UPLC; Waters, Milford, MA, USA) is proposed for the determination of five derivatives of N-acylhomoserine lactones (AHLs) that play a biological role as signal molecules of several gram-negative bacteria. Different commercial SPE cartridges were tested for sample extraction, clean-up and preconcentration. Since the sample matrix was a complex growth media, careful optimization of the SPE with respect to washing procedure, elution solvent and sample solvent was necessary. No sample loss was observed when up to 100 mL spiked full media was added onto the cartridge. Applying UPLC for the determination of AHLs, the performance characteristics of the method showed good separation efficiency and high speed. In order to demonstrate the applicability of the method, supernatants with the known AHL producer Burkholderia cepacia LA3 grown in different media were investigated. Additionally, the method was successfully used for the degradation/uptake study of AHLs from a liquid matrix in which barley was grown under controlled condition.

Original languageEnglish
Pages (from-to)186-193
Number of pages8
JournalJournal of Chromatography A
Volume1134
Issue number1-2
DOIs
StatePublished - 17 Nov 2006
Externally publishedYes

Keywords

  • Method development
  • N-Acylhomoserine lactones
  • Quorum sensing
  • Solid-phase extraction
  • UPLC

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