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4 protocols using microtof q 3 system

1

Synthesis and Characterization of Aromatic Compounds

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3,5-Dimethylphenol, 9-bromophenanthrene, 2-methoxynaphthalene, 3,5-dinitrosalicylaldehyde, and anhydrous magnesium chloride were purchased from TCI Research Chemicals, Haven, Belgium. All other reagents and solvents were analytical grades purchased from Sigma-Aldrich Chemical Co., and used as received unless otherwise stated. Fluka silica gel/TLC-cards 60,778 with fluorescence indicator 254 nm were used for TLC chromatography. Merck silica gel 60 (0.040–0.063 mm, Merck, Darmstadt, Germany was used for flash chromatography purification of the products. 1H-NMR and 13C-NMR spectra were recorded at 500 MHz and 126 MHz or 400 MHz and 100 MHz on an Ultrashield Plus 500 or 400 spectrometer, Bruker, Fallaenden, Germany using CDCl3 or DMSO-d6 as a solvent and TMS as internal standard. LC/MS (ESI) was carried out on a Bruker MicrOTOF-QIII-system with ESI-source with nebulizer 1.2 bar, dry gas 10.0 L/min, dry temperature 220 °C, capillary 4500 V, end plate offset −500 V, Bruker, Hamburg, Germany.
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2

Microbiological and Photophysical Assays

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Terminal alkyne (GFS Chemicals), benzyl bromide and aminoarene reactants (Oakwood Chemical), all other reactants (Aldrich), reaction solvents (Fisher Scientific), and NMR solvents (Cambridge Isotopes) were used as purchased. 2-Azidofluorene,32 (link) 1-azidonaphthalene,33 (link) 2-azidonaphthalene,34 (link) 2-azidoanthracene35 (link) and 1-azidopyrene were33 (link) were prepared by Sandmeyer36 (link) reaction as previously described. Microorganisms were prepared from freeze-dried samples purchased from ATCC (Bacillus subtilus (ATCC 6051), Staphylococcus epidermidis (ATCC 14990), Escherichia coli (ATCC 25922), Klebsiella aerogenes (ATCC 13048), Candida albicans (ATCC 90028) and Saccharomyces cerevisiae (ATCC 9763)). Mueller-Hinton broth and YM broth were purchased from Fisher Scientific and prepared as instructed.
NMR analyses were obtained on a 400 MHz Bruker Ascend system. HRMS analyses were acquired on a Bruker micrOTOF-Q III system using an elution of 0.1% formic acid in methanol. UV-visible absorbance measurements were acquired on an Agilent 8453 spectrophotometer, and data are reported as λmax = nm (log ε). UV-visible emission measurements were acquired on a Varian Cary Eclipse fluorescence spectrophotometer.
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3

Synthesis and Characterization of Isoquinoline Derivatives

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Trimethylsilylacetylene (GFS Chemicals), benzyl bromide and isoquinoline reactants (Oakwood Chemical), Ru and Pd compounds (Oakwood), all other reactants (Aldrich), reaction solvents (Fisher Scientific), and NMR solvents (Cambridge Isotopes) were used as purchased. 1- and 3-isoquinolinyl triflates[27 (link),28 (link)] and 1- and 3-trimethylsilylethynylisoquinolines[29 (link),30 (link)] were prepared as previously described. Microorganisms were prepared from freeze-dried samples purchased from ATCC (Bacillus subtilus (ATCC 6051), Staphylococcus epidermidis (ATCC 14990), Escherichia coli (ATCC 25922), Enterobacter aerogenes (ATCC 13048), Candida albicans (ATCC 90028). Mueller-Hinton broth and YM broth were purchased from Fisher Scientific and prepared as instructed.
NMR analyses were obtained on a 400 MHz Bruker Ascend system. HRMS analyses were acquired on a Bruker micrOTOF-Q III system using an elution of 0.1% formic acid in methanol. UV-visible absorbance measurements were acquired on a Agilent 8453 spectrophotometer, and data are reported as λmax = nm (log ε). Crystals were run on a Rigaku SCX-Mini single-crystal x-ray diffractometer.
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4

CORM-2 Peptide Characterization

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The molar mass of the precursor peptides and the reaction products was measured using a Dionex UltiMate 3000 LC (ThermoScientific, Dreieich, Germany) coupled to a micrOTOF-Q III system (Bruker Daltonics). CORM-2 and peptides 1–7 or Ac-His-NHMe were dissolved in ddH2O and mixed to final concentrations of 12.5 µM and 50 µM, respectively. Samples were incubated at room temperature for ≥15 min prior to application to the LC-MS/MS device.
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