Dec 20, 2014

MALDI-MS-based quantitative analysis for ketone containing homoserine lactones in Pseudomonas aeruginosa

Analytical Chemistry
Yoon-Woo KimYun-Gon Kim

Abstract

N-Acyl homoserine lactones (AHLs), quorum sensing molecules produced by Gram-negative bacteria, are used as important secondary metabolites for antibacterial drug development and cell-to-cell communication. Although various analytical techniques have been developed for detection and quantitation of AHLs from more complex bacterial culture media, only a few methods have been applied to AHL identification in physiological samples. Here, we developed a highly sensitive and reliable MALDI-based 3-oxo AHL quantitation method by employing Girard's reagent T (GT) to produce a permanent cationic charge state [M](+) at the ketone group of AHLs. After extracting AHLs from the supernatant of bacterial cultures using ethyl acetate, the extracts were subsequently derivatized with GT without any additional purification or desalting steps. The chemical derivatization of 3-oxo AHLs dramatically enhanced sensitivity (up to 60 000 times) by lowering the limit of detection (LOD, ∼0.5 fmol)/limit of quantitation (LOQ, ∼2.5 fmol). Additionally, the GT-derivatized 3-oxo AHLs allowed more accurate quantitative analysis from the Pseudomonas aeruginosa PAO1 culture supernatants. This method may be applied for developing high-throughput and sensitive de...Continue Reading

  • References33
  • Citations7

References

  • References33
  • Citations7

Citations

Mentioned in this Paper

Surface Enhanced Laser Desorption Ionization Mass Spectrometry
Pseudomonas aeruginosa (antigen)
Pathogenicity
Homoserine lactone, (S)-isomer
Anterior Horizontal Limb of Lateral Sulcus (Human Only)
Cell Communication
Lactones
Gram-Negative Bacteria
Supernatant
Pseudomonas aeruginosa

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