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题名: 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
作者: Frommberger, M;  Schmitt-Kopplin, P;  Ping, G;  Frisch, H;  Schmid, M;  Zhang, Y;  Hartmann, A;  Kettrup, A
关键词: quorum sensing ;  AHL ;  nano-HPLC ;  sample preconcentration ;  mass spectrometry
刊名: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
发表日期: 2004-02-01
DOI: 10.1007/s00216-003-2400-5
卷: 378, 期:4, 页:1014-1020
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: 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.
关键词[WOS]: CELL-CELL COMMUNICATION ;  LARGE-VOLUME INJECTION ;  L-HOMOSERINE LACTONES ;  PSEUDOMONAS-AERUGINOSA ;  BURKHOLDERIA-CEPACIA ;  STRUCTURAL IDENTIFICATION ;  CAPILLARY ELECTROCHROMATOGRAPHY ;  INTERSPECIES COMMUNICATION ;  STAPHYLOCOCCUS-AUREUS ;  AUTOINDUCER
语种: 英语
WOS记录号: WOS:000189194800020
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/139306
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.GSF Natl Res Ctr Environm & Hlth, Inst Ecol Chem, D-85764 Neuherberg, Germany
2.GSF Natl Res Ctr Environm & Hlth, Inst Soil Ecol, Rhizosphere Biol Grp, D-85764 Neuherberg, Germany
3.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian, Peoples R China
4.Tech Univ Munich, Lehrstuhl Okol Chem & Umweltanalyt, D-8050 Freising Weihenstephan, Germany

Recommended Citation:
Frommberger, M,Schmitt-Kopplin, P,Ping, G,et al. 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[J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY,2004,378(4):1014-1020.
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