title: Determination of autoinducer-2 in biological samples by high-performance liquid chromatography with fluorescence detection using pre-column derivatization.
abstract: Autoinducer-2 (AI-2), as a small-molecular-weight organic molecule secreted and perceived by various bacteria, enables intra- and inter-species communications. Quantitative determination of AI-2 is essential for exploring the bacterial AI-2-related physiological and biochemical processes. However, current strategies for sensitive detection of AI-2 require sophisticated instruments and complicated procedures. In this work, on the basis of the derivatization of AI-2 with 2,3-diaminonaphthalene, a simple, sensitive and cost-effective high-performance liquid chromatography with fluorescence detector (HPLC-FLD) method is developed for the quantitative detection of AI-2. Under the optimized conditions, this method had a broad linear range of 10-14,000 ng/ml (R(2)=0.9999), and a low detection limit of 1.0 ng/ml. Furthermore, the effectiveness of this approach was further validated through measuring the AI-2 concentrations in the cell-free culture supernatants of both Escherichia coli and Vibrio harveyi.
authors.affiliation: Department of Chemistry, University of Science & Technology of China, Hefei 230026, China.
authors.forename: Xiang-Ning
authors.initials: XN
authors.lastname: Song
authors.affiliation: Department of Chemistry, University of Science & Technology of China, Hefei 230026, China.
authors.forename: Hai-Bin
authors.initials: HB
authors.lastname: Qiu
authors.affiliation: Department of Chemistry, University of Science & Technology of China, Hefei 230026, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Xiao
authors.affiliation: Department of Chemistry, University of Science & Technology of China, Hefei 230026, China.
authors.forename: Yuan-Yuan
authors.initials: YY
authors.lastname: Cheng
authors.affiliation: Department of Chemistry, University of Science & Technology of China, Hefei 230026, China.
authors.forename: Wen-Wei
authors.initials: WW
authors.lastname: Li
authors.affiliation: Department of Chemistry, University of Science & Technology of China, Hefei 230026, China. Electronic address: gpsheng@ustc.edu.cn.
authors.forename: Guo-Ping
authors.initials: GP
authors.lastname: Sheng
authors.affiliation: Environmental Engineering Research Centre, The University of Hong Kong, Hong Kong, China.
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Li
authors.affiliation: Department of Chemistry, University of Science & Technology of China, Hefei 230026, China. Electronic address: hqyu@ustc.edu.cn.
authors.forename: Han-Qing
authors.initials: HQ
authors.lastname: Yu
mesh_terms: D002474:Cell-Free System / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry*; D006714:Homoserine / Q000031:analogs & derivatives* / Q000032:analysis; D007783:Lactones / Q000032:analysis*; D057230:Limit of Detection; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*; D014733:Vibrio / Q000737:chemistry*
chemical_list: D007783:Lactones; C091088:N-octanoylhomoserine lactone; D006714:Homoserine
keywords: 2,3-Diaminonaphthalene; Autoinducer-2; Biological samples; Detection; HPLC–FLD