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A simple and robust set-up for on-column sample preconcentration - nano-liquid chromatography - electrospray ionization mass spectrometry for the analysis of N-acylhomoserine lactones.

Anal. Bioanal. Chem. 378, 1014-1020 (2004)
DOI
Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
A simple method for the simultaneous, rapid and sensitive determination of N-acylhomoserine lactone signaling molecules in bacterial isolates, without prior sample preconcentration and with minimal sample cleanup, is presented. The analysis relies on the combination of analyte preconcentration and separation on a single device: a relatively large sample volume (1-5 muL) is directly loaded onto a laboratory-made, miniaturized (75 mum i.d.) reverse phase nano-liquid chromatography column, connected on-line to a microelectrospray-ionization ion trap mass spectrometer. In a first step the analyte is adsorbed (and so concentrated) at the beginning of the column, and is eluted and selectively separated in a second step by the organic mobile phase. Sample preconcentration follows the mechanisms of solid phase extraction on a nano-scale, while separation takes place according to classical liquid chromatography separation principles. The columns can be manufactured easily, are simply connected, and used with minimal solvent amounts; this makes this method extremely robust and cost-effective. The analytical setup was found to be routinely quantitative down to a concentration of 10 ng/mL (corresponding to a total analyte amount of 10 pg or ca. 50 fmol). The limit of detection was reached at 1 ng/mL (1 pg, ca. 5 fmol). Compared to the classical AHL analysis of bacterial cultures with biosensors, where selectivity and sensitivity is often limited, this rapid analytical technique is a substantial qualitative and quantitative improvement. Two unsubstituted N-acylhomoserine lactones could be identified and quantified from a Burkholderia cepacia culture supernatant in a chloroform extract.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Quorum sensing; AHL; Nano-HPLC; Sample preconcentration; Mass spectrometry
ISSN (print) / ISBN 1618-2642
e-ISSN 1618-2650
Quellenangaben Band: 378, Heft: 4, Seiten: 1014-1020 Artikelnummer: , Supplement: ,
Verlag Springer
Verlagsort Heidelberg
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Soil Ecology (IBOE)
Institute of Ecological Chemistry (IOEC)