Mass Electrospray ionization mass spectrometry (ESI-MS) was performed on the AmazonX mass spectrometer (Bruker Daltonik GmbH, Bremen, Germany). The measurements were carried out in the positive/negative ion detection mode in the m/z range from 100 to 1000. The voltage on the capillary was 140 V. Data were processed using the DataAnalysis 4.0 program (Bruker Daltonik GmbH, Bremen, Germany).
Amazonx mass spectrometer
Manufactured by Bruker
Sourced in Germany
The AmazonX is a mass spectrometer manufactured by Bruker. It is designed to identify and quantify a wide range of chemical compounds with high accuracy and sensitivity.
Automatically generated - may contain errors
Lab products found in correlation
Dataanalysis 4.0 program, by Bruker (2 mentions)
Dataanalysis 4, by Bruker (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Pyrene, by Merck Group (1 mentions)
Dionex ultimate 3000 system, by Thermo Fisher Scientific (1 mentions)
Acquity uplc beh c18 column, by Waters Corporation (1 mentions)
Ultimate 3000, by Bruker (1 mentions)
6 protocols using amazonx mass spectrometer
1
Mass Spectrometry Analysis of Biomolecules
2
ESI-MS Analysis of Organic Compounds
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Electrospray ionization mass spectra (ESI) were obtained on an AmazonX mass spectrometer (Bruker Daltonik GmbH, Bremen, Germany). The measurements were carried out in the regime of positive ions in the range of m/z from 100 to 2800. The voltage on the capillary was −4500 V. Nitrogen was used as the gas-drier with a temperature of 300 °C and a flow rate of 10 L∙min−1. The compounds were dissolved in acetonitrile to a concentration of 10−6 g/L. The data was processed using DataAnalysis 4.0 (Bruker Daltonik GmbH, Bremen, Germany).
3
Synthesis and Characterization of Calix[4]resorinarenes
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The syntheses of compounds 1 and 2 and their 1H NMR, IR and mass spectra are presented in Supplementary Materials . The initial calix[4]resorinarenes with pentyl and undecyl substituents on the lower rim were synthesized as described in ref. [54 (link)]. The syntheses of the compounds 3 –6 were first reported in this study. BSA (≥96.0%) and pyrene (99%) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The stock solution of 10.0 × 10−6 mol L−1 BSA was prepared in 0.1 mol L−1 phosphate buffer solution (PB) (pH = 7.0). The stock solution of pyrene was prepared in ethanol at a concentration of 9.71 × 10−4 mol L−1.
1H and 13C NMR spectra were performed on an MSL-400 spectrometer (400 MHz for 1H NMR, 100 MHz for 13C NMR). The chemical shifts were reported relative to the residual solvent peaks as internal standards. IR spectra were recorded with a Vector 22 Spectrometer in KBr pellets. Electrospray ionization mass spectra (ESI) were obtained on an AmazonX mass spectrometer (Bruker Daltonik GmbH, Bremen, Germany).
1H and 13C NMR spectra were performed on an MSL-400 spectrometer (400 MHz for 1H NMR, 100 MHz for 13C NMR). The chemical shifts were reported relative to the residual solvent peaks as internal standards. IR spectra were recorded with a Vector 22 Spectrometer in KBr pellets. Electrospray ionization mass spectra (ESI) were obtained on an AmazonX mass spectrometer (Bruker Daltonik GmbH, Bremen, Germany).
4
Metabolomic Analysis of Bacterial Extracts
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PaATCC43949 was grown at 28°C and 37°C in LB medium upon addition of 10% (w/v) amberlite XAD-16 adsorber resin. Supernatant and XAD-16 were separated by decantation. After washing the adsorber resin the bound compounds were eluted upon incubating twice with 2 ml methanol at room temperature for 20 min and the solvent was removed under reduced pressure. Then the samples were re-dissolved in 0.5 ml methanol, centrifuged (10 min, 13500 rpm, room temperature), diluted 1:10 with methanol and analysed with a Dionex Ultimate 3000 system (Thermo Scientific, Dreieich, Germany) using an Acquity UPLC BEH C18 1.7μm RP column (Waters GmbH, Eschborn, Germany) coupled to an AmaZon X mass spectrometer (Bruker Daltonik GmbH, Bremen, Germany). Metabolites were gradually eluted from the column in 22 min (gradient from 5% to 95% acetonitrile in H2O with 0.1% formic acid; flowrate of 0.6 mL/min).
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PaATCC43949 was grown at 28°C and 37°C in LB medium upon addition of 10% (w/v) amberlite XAD-16 adsorber resin. Supernatant and XAD-16 were separated by decantation. After washing the adsorber resin the bound compounds were eluted upon incubating twice with 2 ml methanol at room temperature for 20 min and the solvent was removed under reduced pressure. Then the samples were re-dissolved in 0.5 ml methanol, centrifuged (10 min, 13500 rpm, room temperature), diluted 1:10 with methanol and analysed with a Dionex Ultimate 3000 system (Thermo Scientific, Dreieich, Germany) using an Acquity UPLC BEH C18 1.7μm RP column (Waters GmbH, Eschborn, Germany) coupled to an AmaZon X mass spectrometer (Bruker Daltonik GmbH, Bremen, Germany). Metabolites were gradually eluted from the column in 22 min (gradient from 5% to 95% acetonitrile in H2O with 0.1% formic acid; flowrate of 0.6 mL/min).
5
Mass Spectrometry Characterization
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Mass Electrospray ionization mass spectrometry (ESI-MS) was performed on the AmazonX mass spectrometer (Bruker Daltonics, Bremen, Germany). The measurements were carried out in the positive/negative ion detection mode in the m/z range from 100 to 1000. The voltage on the capillary was 140 V. Data was processed using the DataAnalysis 4.0 program (Bruker Daltonics).
6
Bacterial Metabolite Extraction and Analysis
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The bacterial
cell pellets and XAD beads were collected after centrifugation and
resuspended in 10 mL of methanol. XAD beads were washed with methanol
by inverting for 1 h, followed by separating from methanol through
filter paper. The resulting methanol extracts were evaporated to dryness
and redissolved in 1 mL of fresh methanol. The methanol extracts obtained
above (1 mL) were cleaned up via centrifugation at 17 000g for 20 min. Twenty microliters of crude extracts was diluted
in 180 μL of methonal before analysis, 5 μL of which was
injected and analyzed by HPLC-ESI-MS by a Dionex UltiMate 3000 system
coupled to a Bruker AmaZon X mass spectrometer or HPLC-ESI-HRMS (Impact
II) using an ACQUITY UPLC BEH C18 column (130 Å, 2.1 mm ×
100 mm, 1.7 μm particle size, Waters GmbH) at a flow rate of
0.6 mL/min using acetonitrile and water containing 0.1% formic acid
(v/v) in a gradient ranging from 5 to 95% of acetonitrile (ACN) over
16 min. Spectra for RXPs were recorded in a positive ion mode with
the range from 100 to 1200 m/z and UV at 200–600
nm.
cell pellets and XAD beads were collected after centrifugation and
resuspended in 10 mL of methanol. XAD beads were washed with methanol
by inverting for 1 h, followed by separating from methanol through
filter paper. The resulting methanol extracts were evaporated to dryness
and redissolved in 1 mL of fresh methanol. The methanol extracts obtained
above (1 mL) were cleaned up via centrifugation at 17 000g for 20 min. Twenty microliters of crude extracts was diluted
in 180 μL of methonal before analysis, 5 μL of which was
injected and analyzed by HPLC-ESI-MS by a Dionex UltiMate 3000 system
coupled to a Bruker AmaZon X mass spectrometer or HPLC-ESI-HRMS (Impact
II) using an ACQUITY UPLC BEH C18 column (130 Å, 2.1 mm ×
100 mm, 1.7 μm particle size, Waters GmbH) at a flow rate of
0.6 mL/min using acetonitrile and water containing 0.1% formic acid
(v/v) in a gradient ranging from 5 to 95% of acetonitrile (ACN) over
16 min. Spectra for RXPs were recorded in a positive ion mode with
the range from 100 to 1200 m/z and UV at 200–600
nm.
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