Quantitative determination of AI-2 quorum-sensing signal of bacteria using high performance liquid chromatography–tandem mass spectrometry  

作　　者：Fang Xu Xiangning Song Peijie Cai Guoping Sheng Hanqing Yu 

机构地区：[1]CAS Key Laboratory of Urban Pollutant Conversion, Department of Chemistry, University of Science & Technology of China [2]School of Medical Engineering, Hefei University of Technology

出　　处：《Journal of Environmental Sciences》2017年第2期204-209,共6页环境科学学报（英文版）

基　　金：supported by the National Natural Science Foundation of China(Nos.21261160489,21207031 and51538011);the Program for Changjiang Scholars and Innovative Research Team in University,China

摘　　要：Autoinducer 2（AI-2）, an important bioactive by-product of the Lux S-catalyzed S-ribosylhomocysteine cleavage reaction in the activated-methyl-cycle, has been suggested to serve as a universal intra- and inter-species signaling molecule. The development of reliable and sensitive methods for quantitative determination of AI-2 is highly desired.However, the chemical properties of AI-2 cause difficulty in its quantitative analysis.Herein, we report a high performance liquid chromatography-tandem mass spectrometric method that enables reproducible and sensitive measurement of AI-2 concentrations in complex matrixes. 4,5-Dimethylbenzene-1,2-diamine（DMBDM）, an easy-to-obtain commercial reagent, was used for the derivatization treatment. The assay was linear in the concentration range of 1.0–1000 ng/m L（R^2= 0.999） and had a lower limit of quantification of0.58 ng/m L. The method exhibited several advantages, e.g., high selectivity, wide linear response range, and good sensitivity. Furthermore, the effectiveness of the method was further validated through measuring AI-2 concentrations in the cell-free culture supernatant from Escherichia coli wild type.Autoinducer 2（AI-2）, an important bioactive by-product of the Lux S-catalyzed S-ribosylhomocysteine cleavage reaction in the activated-methyl-cycle, has been suggested to serve as a universal intra- and inter-species signaling molecule. The development of reliable and sensitive methods for quantitative determination of AI-2 is highly desired.However, the chemical properties of AI-2 cause difficulty in its quantitative analysis.Herein, we report a high performance liquid chromatography-tandem mass spectrometric method that enables reproducible and sensitive measurement of AI-2 concentrations in complex matrixes. 4,5-Dimethylbenzene-1,2-diamine（DMBDM）, an easy-to-obtain commercial reagent, was used for the derivatization treatment. The assay was linear in the concentration range of 1.0–1000 ng/m L（R^2= 0.999） and had a lower limit of quantification of0.58 ng/m L. The method exhibited several advantages, e.g., high selectivity, wide linear response range, and good sensitivity. Furthermore, the effectiveness of the method was further validated through measuring AI-2 concentrations in the cell-free culture supernatant from Escherichia coli wild type.

关 键 词：Autoinducer-2 Quorum sensing Signal transduction HPLC–MS/MS Quantitative analysis Escherichia coli wild type 

分 类 号：Q503[生物学—生物化学]