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Realised by ALMS™
developer of the AIDS-HIV Reference project
Abstract No.: ThP-LB4
Session: LATE-BREAKING/Environmental Analysis
Presentation date: Thu, Aug 31, 2006
Presentation time: 14:30 – 16:00

Confirmation of the Occurrence of N-acylhomoserine Lactones (HSL) in Different Real Samples by Combined Results of FTICR/MS Measurements, UPLC Analysis, In Silico Predictions and by Application of Biosensor Constructs

Matthias Englmann1, Agnes Fekete1, Michael Rothballer2, Moritz Frommberger1, Xiaojing Li1,5, Jenoe Fekete3, Anton Hartmann2, Leo Eberl4, Philippe Schmitt-Kopplin1

1 Institute of Ecological Chemistry, GSF National Research Center, Neuherberg, Germany
2 Institute of Soil Ecology, GSF National Research Center, Neuherberg, Germany
3 Dep. of General and Analytical Chemistry, Budapest University of Technology, Budapest, Hungary
4 Department of Microbiology, University Zurich, Zurich, Switzerland
5 Department of Chemistry, Fuzhou University, Fujian, China

Correspondence address: Matthias Englmann, GSF National Research Center for Environment and Health, Institute of Ecological Chemistry, Ingolstadter Landstrasse 1, Neuherberg, D-85764 Germany.

Keywords: Chromatography, Liquid -, High Pressure (HPLC); Environmental; Esters, Fatty Acid; Mass Spectrometry, Ion Cyclotron Resonance.

Novel aspect: A combination of four independent approaches was used for the confirmation of the occurrence of signalling molecules in different real samples.

 

Four approaches based on different principles are presented, which, when used in combination with each other, give information for the identification of N-acyl-homoserine lactones (HSL) in real samples. HSL are important signalling molecules involved in cell to cell communications of gram negative bacteria.
An UPLC-DAD analysis following a SPE cleanup and concentration step results in 10 to 50 peaks which could correspond to target-compounds. UV-Vis absorbtion spectra give further weak hints to encircle 1 to 5 of them.
An in silico prediction of the UPLC method which is applicable for 26 compounds, based on the chemical properties of the analytes, encircles 5-20 candidates out of the UPLC measurement done before.
FTICR/MS gives up to 10,000 resolved masses. 3 to 50 of them represent elemental compositions possibly fitting the chemical structure of target analytes.
The intersection of these approaches results in 0 to 5 candidates, which are in a fourth step confirmed by thin layer chromatography with detection by reporter strains.