Development and application of a method for the analysis of N-acylhomoserine lactones by solid-phase extraction and ultra high pressure liquid chromatography.
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.
Li
Xiaojing
X
Ministry of Education Key Laboratory of Analysis and Detection Technology for Food Safety, and Department of Chemistry, Fuzhou University, Fuzhou, Fujian 350002, China.
Fekete
Agnes
A
Englmann
Matthias
M
Götz
Christine
C
Rothballer
Michael
M
Frommberger
Moritz
M
Buddrus
Katharina
K
Fekete
Jenoe
J
Cai
Chunping
C
Schröder
Peter
P
Hartmann
Anton
A
Chen
Guonan
G
Schmitt-Kopplin
Philippe
P
D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000032:analysis / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D016956:Burkholderia cepacia; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D001467:Hordeum; D052616:Solid Phase Extraction / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet
C088386:homoserine lactone; D015107:4-Butyrolactone