TY - DATA T1 - MALDI-MS-Based Quantitative Analysis for Ketone Containing Homoserine Lactones in Pseudomonas aeruginosa PY - 2015/01/20 AU - Yoon-Woo Kim AU - Changmin Sung AU - Seulee Lee AU - Kyoung-Jin Kim AU - Yung-Hun Yang AU - Byung-Gee Kim AU - Yoo Kyung Lee AU - Hee Wook Ryu AU - Yun-Gon Kim UR - https://acs.figshare.com/articles/journal_contribution/MALDI_MS_Based_Quantitative_Analysis_for_Ketone_Containing_Homoserine_Lactones_in_i_Pseudomonas_aeruginosa_i_/2214343 DO - 10.1021/ac5039362.s001 L4 - https://ndownloader.figshare.com/files/3849898 KW - Ketone Containing Homoserine Lactones KW - virulence factor characterization KW - GT KW - drug development KW - AHL KW - oxo KW - detection KW - method KW - Pseudomonas aeruginosa PAO 1 culture supernatants KW - LOD KW - LOQ KW - 60 000 times KW - quantitation KW - cationic charge state N2 - N-Acyl homoserine lactones (AHLs), quorum sensing molecules produced by Gram-negative bacteria, are used as important secondary metabolites for antibacterial drug development and cell-to-cell communication. Although various analytical techniques have been developed for detection and quantitation of AHLs from more complex bacterial culture media, only a few methods have been applied to AHL identification in physiological samples. Here, we developed a highly sensitive and reliable MALDI-based 3-oxo AHL quantitation method by employing Girard’s reagent T (GT) to produce a permanent cationic charge state [M]+ at the ketone group of AHLs. After extracting AHLs from the supernatant of bacterial cultures using ethyl acetate, the extracts were subsequently derivatized with GT without any additional purification or desalting steps. The chemical derivatization of 3-oxo AHLs dramatically enhanced sensitivity (up to 60 000 times) by lowering the limit of detection (LOD, ∼0.5 fmol)/limit of quantitation (LOQ, ∼2.5 fmol). Additionally, the GT-derivatized 3-oxo AHLs allowed more accurate quantitative analysis from the Pseudomonas aeruginosa PAO1 culture supernatants. This method may be applied for developing high-throughput and sensitive detection methods of quorum sensing signal molecules in biofilm-related clinical applications such as virulence factor characterization and antibacterial drug development. ER -