1H and 13C NMR spectra were collected on a Bruker AVANCE III spectrometer (Germany) at 500 and 125 MHz, respectively, and chemical shifts are given in δ (ppm) using CDCl3, CD3OD, (CD3)2CO, or (CD3)2SO as solvent. High-resolution mass spectrometry (HRMS) analysis was performed using a Bruker MicroTOF-QII-system with an ESI-source. Melting point was determined using a Bibby Stuart Melting point SMB3 (UK). UV spectra were measured in ethanol on a T92+ Spectrophotometer (PG instrument, UK). Merck precoated silica gel plates (Kieselgel 60 F254, Germany) were used for analytical thin-layer chromatography (TLC). Merck preparative layer chromatography plates (PLC silica gel 60 F254, 2 mm thickness) and Scharlau silica gel 60 (0.06-0.2 mm) (Spain) were used for isolation and purification.
Microtof q 2 system
Manufactured by Bruker
Sourced in United States
The MicrOTOF-Q II system is a high-performance quadrupole time-of-flight (Q-TOF) mass spectrometer designed for accurate mass determination and high-resolution analysis of small molecules and biomolecules. It features a hybrid quadrupole-time-of-flight mass analyzer that combines the selectivity of a quadrupole with the high resolving power and mass accuracy of a time-of-flight mass analyzer.
Automatically generated - may contain errors
Lab products found in correlation
Compass dataanalysis 4, by Bruker (2 mentions)
Esi tof tuning mix calibrant, by Merck Group (2 mentions)
Avance 3 spectrometer, by Bruker (1 mentions)
Kieselgel 60 f254, by Merck Group (1 mentions)
Plc silica gel 60 f254, by Scharlab (1 mentions)
C18 pre column, by Thermo Fisher Scientific (1 mentions)
Quantanalysis, by Bruker (1 mentions)
5 protocols using microtof q 2 system
1
Spectroscopic Characterization of Compounds
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1H and 13C NMR spectra were collected on a Bruker AVANCE III spectrometer (Germany) at 500 and 125 MHz, respectively, and chemical shifts are given in δ (ppm) using CDCl3, CD3OD, (CD3)2CO, or (CD3)2SO as solvent. High-resolution mass spectrometry (HRMS) analysis was performed using a Bruker MicroTOF-QII-system with an ESI-source. Melting point was determined using a Bibby Stuart Melting point SMB3 (UK). UV spectra were measured in ethanol on a T92+ Spectrophotometer (PG instrument, UK). Merck precoated silica gel plates (Kieselgel 60 F254, Germany) were used for analytical thin-layer chromatography (TLC). Merck preparative layer chromatography plates (PLC silica gel 60 F254, 2 mm thickness) and Scharlau silica gel 60 (0.06-0.2 mm) (Spain) were used for isolation and purification.
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1H and 13C NMR spectra were collected on a Bruker AVANCE III spectrometer (Germany) at 500 and 125 MHz, respectively, and chemical shifts are given in δ (ppm) using CDCl3, CD3OD, (CD3)2CO, or (CD3)2SO as solvent. High-resolution mass spectrometry (HRMS) analysis was performed using a Bruker MicroTOF-QII-system with an ESI-source. Melting point was determined using a Bibby Stuart Melting point SMB3 (UK). UV spectra were measured in ethanol on a T92+ Spectrophotometer (PG instrument, UK). Merck precoated silica gel plates (Kieselgel 60 F254, Germany) were used for analytical thin-layer chromatography (TLC). Merck preparative layer chromatography plates (PLC silica gel 60 F254, 2 mm thickness) and Scharlau silica gel 60 (0.06-0.2 mm) (Spain) were used for isolation and purification.
2
Nano LC-ESI-QTOF-MS/MS Peptide Analysis
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In-gel digested peptides were analyzed by nano LC-ESI-QTOF-MS/MS on a Bruker micrOTOF-Q II system. For LC-MS/MS analysis 15 μL of each sample were injected. Peptides were first trapped and preconcentrated on a C-18 precolumn (Dionex) at 30 μL flow for 5 min and later eluted on the separation column with a flow of 220 nL/min (column dimensions were I.D. 75 μm, length 15 cm, PepMap C-18, 3 μm, 100 Å). Solvents used for elution of peptides were solvent A: water, ACN (1.0%), and formic acid (0.1%) and solvent B: ACN and formic acid (0.1%). All samples were measured in “auto” MS/MS mode, positive ion mode on the Bruker nanospray source with a capillary voltage 1,500 Volts, dry gas flow 6.0 L/min, dry temperature: 130°C, 1 MS followed by 5 MS/MS of the most intense ions, total cycle time 4.4–8.8 sec, m/z 400–1,400 taken as precursor ions for MS/MS, active exclusion after 2 spectra for 0.5 min, and threshold for MS/MS set to 1,000.
3
DIESI-MS Analysis of Metabolite Samples
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DIESI-MS analysis was done on a Bruker MicrOTOF-QII system, using an electrospray ionization (ESI) interface (Bruker Daltonics, Billerica, MA, USA) operated in the negative ion mode. A solution of 10 µL of the sample resuspended in 1 mL of methanol was filtered with a 0.25 µm polytetrafluoroethylene (PTFE) filter and diluted 1:100 with methanol. Diluted samples were directly infused into the ESI source and analyzed in negative mode. Nitrogen was used with a flow rate of 4 L/min (0.4 Bar) as a drying and nebulizer gas, with a gas temperature of 180 °C and a capillary voltage set to −4500 V. The spectrometer was calibrated with an ESI-TOF tuning mix calibrant (Sigma-Aldrich, Toluca, Estado de México, Mexico).
MS/MS analysis was performed using positive and negative electrospray ionization (ESI+/−), and the obtained fragments were analyzed by a Bruker Compass Data Analysis 4.0 (Bruker Daltonics, Technical Note 008, 2004, Billerica, MA, USA). An accuracy threshold of 5 ppm was established to confirm the elemental compositions.
MS/MS analysis was performed using positive and negative electrospray ionization (ESI+/−), and the obtained fragments were analyzed by a Bruker Compass Data Analysis 4.0 (Bruker Daltonics, Technical Note 008, 2004, Billerica, MA, USA). An accuracy threshold of 5 ppm was established to confirm the elemental compositions.
4
Direct Ionization Mass Spectrometry of Samples
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Direct Ionization analysis (DIESI-MS) was done on a Bruker MicrOTOF-QII system, using an electrospray ionization (ESI) interface (Bruker Daltonics, Biellerica, MA, USA) operated in the negative ion mode. A solution of 10 µL of the sample resuspended in 1 mL of methanol was filtered with a 0.25 µm polytetrafluoroethylene (PTFE) filter and diluted 1:100 with methanol. Diluted samples were directly infused into the ESI source and analyzed in negative mode. Nitrogen was used with a flow rate of 4 L/min (0.4 Bar) as a drying and nebulizer gas, with a gas temperature of 180 °C and a capillary voltage set to 4500 V. The spectrometer was calibrated with an ESI-TOF tuning mix calibrant (Sigma-Aldrich, Toluca, Estado de México, México).
MS/MS analysis was performed using negative electrospray ionization (ESI−), and the obtained fragments were analyzed by a Bruker Compass Data Analysis 4.0 (Bruker Daltonics, Technical Note 008, 2004, Bruker Daltonics, Biellerica, MA, USA). An accuracy threshold of 5 ppm was established to confirm the elemental compositions.
MS/MS analysis was performed using negative electrospray ionization (ESI−), and the obtained fragments were analyzed by a Bruker Compass Data Analysis 4.0 (Bruker Daltonics, Technical Note 008, 2004, Bruker Daltonics, Biellerica, MA, USA). An accuracy threshold of 5 ppm was established to confirm the elemental compositions.
5
Quantitative Analysis of Targeted Metabolites
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Samples were ground in liquid nitrogen and aliquoted to 10 – 100 mg depending on the tissues and their known DTG and flavonoid concentrations (precise mass was recorded). Approximately 100 mg of aliquoted samples were extracted using 1 mL 80% methanol aqueous buffer and analyzed on a micrOTOF-Q II system (Bruker Daltonics) as previously described (Heiling et al., 2016 (link); Li et al., 2016 (link)). QuantAnalysis (Bruker Daltonics) software was used to integrate the DTG peak areas based on each compound’s diagnostic m/z value and retention time as described in Figure 1—figure supplement 1 .
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