Bioanalysis of Pseudomonas aeruginosa alkyl quinolone signalling molecules in infected mouse tissue using LC–MS/MS; and its application to a pharmacodynamic evaluation of MvfR inhibition

Turnpenny, Paul; Padfield, Anthony; Barton, Patrick; Teague, Joanne; Rahme, Laurence G.; Pucci, Michael J.; Zahler, Robert; Rubio, Aileen

doi:10.1016/j.jpba.2017.02.034

Cited by 12 publications

(23 citation statements)

References 27 publications

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“…Since the analysis of bacterial cultures and plasma samples of patients were completed only 2 days after sampling and handling, and samples were stored refrigerated at −4°C, no other pathogens should develop [33]. In this situation, the unknown AHLs and HQs detected with the developed MS methods resulted from P. aeruginosa bacteria or patients with sepsis.…”

Section: Sample Results Discussionmentioning

confidence: 99%

“…Chromatographic separation for the MRM and PI analysis of C7 HQ in bacterial cultures and plasma samples was achieved using 2-picolinic acid 2 mM/formic acid 0.1% in water (solvent C) and acetonitrile 0.1% formic acid (solvent D). 2-Picolinic acid acted as a bidentate chelator, preventing peak distortion caused by C7 HQ, an iron chelator molecule [33]. Gradient started from 20% solvent D, up to 100% solvent D in 12 min; then the column returned to the starting conditions.…”

Section: Hplc Parametersmentioning

confidence: 99%

“…As described above, AHLs and HQs are the most abundant QS signalling molecules produced by Gram-negative bacteria. Many studies in the literature have aimed to describe and quantify well-known or new QS molecules in bacterial culture or human samples [20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35]. There are two main methods for obtaining the measurements: biosensors [21-23, 26, 30, 31] and methods based on liquid chromatography coupled to (high-resolution) mass spectrometry [20, 23-25, 27-29, 32-36].…”

Section: Introductionmentioning

confidence: 99%

“…During the past few years, many LC-MS methods have been developed that exploit several stationary and mobile phases, using both targeted and untargeted MS approaches. Some studies showed a very low limit of quantitation (pg/mL) and good selectivity, optimizing a short liquid chromatography separation both for AHLs, furthermore enantiomeric, and for HQs [27][28][29][33][34][35][36]. Other research groups have developed extended chromatography separation runs to provide the quantitation of a higher number of molecules [24] or to identify new AI molecules with an untargeted highresolution MS approach [32].…”

Section: Introductionmentioning

confidence: 99%

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Targeted and untargeted quantification of quorum sensing signalling molecules in bacterial cultures and biological samples via HPLC-TQ MS techniques

et al. 2020

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Quorum sensing (QS) is the ability of some bacteria to detect and to respond to population density through signalling molecules. QS molecules are involved in motility and cell aggregation mechanisms in diseases such as sepsis. Few biomarkers are currently available to diagnose sepsis, especially in high-risk conditions. The aim of this study was the development of new analytical methods based on liquid chromatography-mass spectrometry for the detection and quantification of QS signalling molecules, including N-acyl homoserine lactones (AHL) and hydroxyquinolones (HQ), in biofluids. Biological samples used in the study were Pseudomonas aeruginosa bacterial cultures and plasma from patients with sepsis. We developed two MS analytical methods, based on neutral loss (NL) and product ion (PI) experiments, to identify and characterize unknown AHL and HQ molecules. We then established a multiple-reaction-monitoring (MRM) method to quantify specific QS compounds. We validated the HPLC-MS-based approaches (MRM-NL-PI), and data were in accord with the validation guidelines. With the NL and PI MS-based methods, we identified and characterized 3 and 13 unknown AHL and HQ compounds, respectively, in biological samples. One of the newly found AHL molecules was C12-AHL, first quantified in Pseudomonas aeruginosa bacterial cultures. The MRM quantitation of analytes in plasma from patients with sepsis confirmed the analytical ability of MRM for the quantification of virulence factors during sepsis.

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Section: Sample Results Discussionmentioning

confidence: 99%

Section: Hplc Parametersmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Targeted and untargeted quantification of quorum sensing signalling molecules in bacterial cultures and biological samples via HPLC-TQ MS techniques

et al. 2020

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“…This communication network, known as quorum sensing (QS), regulates different phenotypes including antibacterial resistance, bacterial motility, biofilm development, and expression of virulence factors and toxins. [ 11 ]…”

Section: Introductionmentioning

confidence: 99%

Synthesis of imidazolidine‐2,4‐dione and 2‐thioxoimidazolidin‐4‐one derivatives as inhibitors of virulence factors production in Pseudomonas aeruginosa

Mohamed

Abdel‐Samii

Abdel‐Aal

et al. 2020

Archiv der Pharmazie

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In an attempt to counteract bacterial pathogenicity, a set of novel imidazolidine-2,4dione and 2-thioxoimidazolidin-4-one derivatives was synthesized and evaluated as inhibitors of bacterial virulence. The new compounds were characterized and screened for their effects on the expression of virulence factors of Pseudomonas aeruginosa, including protease, hemolysin, and pyocyanin. Imidazolidine-2,4-diones 4c, 4j, and 12a showed complete inhibition of the protease enzyme, and they almost completely inhibited the production of hemolysin at 1/4 MIC (1/4 minimum inhibitory concentration; 1, 0.5, and 0.5 mg/ml, respectively). 2-Thioxoimidazolidin-4-one derivative 7a exhibited the best inhibitory activity (96.4%) against pyocyanin production at 1 mg/ml (1/4 MIC). A docking study was preformed to explore the potential binding interactions with quorum-sensing receptors (LasR and RhlR), which are responsible for the expression of virulence genes. K E Y W O R D S2-thioxoimidazolidin-4-one, imidazolidine-2,4-dione, Pseudomonas aeruginosa, quorum sensing, virulence inhibition

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Deciphering and investigating fragment mechanism of quinolones using multi‐collision energy mass spectrometry and computational chemistry strategy

Lin

Zhou

Zhang

et al. 2023

Rapid Comm Mass Spectrometry

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Rationale: Quinolones show characteristic fragments in mass spectrometry (MS) analysis due to their common core structures, and energy-dependent differences among these fragments are generated through the same fragmentation pathway of different molecules. Computational chemistry, which provides quantitative results of molecule parameters, is helpful for investigating the mechanisms of chemistry.Methods: MS/MS spectra of five quinolones, namely norfloxacin (NOR), enoxacin (ENO), enrofloxacin (ENR), gatifloxacin (GAT), and lomefloxacin (LOM), were acquired for deciphering fragmentation pathways under multi-collision energy (CE).Computational methods were used for excluding little possibility pathways from the point of view of energy and stable conformations, whereas optimized collision energy (OCE) and maximum relative intensity (MRI) of major competitive fragments were investigated and confirmed using computational results.Results: Fragmentation results of NOR, ENO, ENR, and GAT were deciphered using experimental and computational data, of which fragmentation regularities were summarized. Fragmentation pathways of LOM were deciphered under the guidance of foregoing regularities. Meanwhile, the whole process was validated by comparing OCE and MRI and computational energy results, which showed good agreement.Conclusions: A strategy for explaining quinolone fragmentation results of multi-CE values and deciphering fragment mechanism using computational methods was developed. Relevant data and strategy may provide ideas for how to design and decipher new drug molecules with similar structures.

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Bioanalysis of Pseudomonas aeruginosa alkyl quinolone signalling molecules in infected mouse tissue using LC–MS/MS; and its application to a pharmacodynamic evaluation of MvfR inhibition

Cited by 12 publications

References 27 publications

Targeted and untargeted quantification of quorum sensing signalling molecules in bacterial cultures and biological samples via HPLC-TQ MS techniques

Targeted and untargeted quantification of quorum sensing signalling molecules in bacterial cultures and biological samples via HPLC-TQ MS techniques

Synthesis of imidazolidine‐2,4‐dione and 2‐thioxoimidazolidin‐4‐one derivatives as inhibitors of virulence factors production in Pseudomonas aeruginosa

Deciphering and investigating fragment mechanism of quinolones using multi‐collision energy mass spectrometry and computational chemistry strategy

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