Approximately 50 mg of frozen heart was grinded in a mortar submerged in liquid nitrogen. Metabolites were extracted using a solution with 50% methanol, 20% acetonitrile and 30% H2O. LC-MS analysis of the extracted samples was performed using a Q Exactive Hybrid Quadrupole-Orbitrap mass spectrometer coupled to a Vanquish UHPLC system (Thermo Fisher Scientific). The liquid chromatography system was fitted with a Sequant ZIC-pHILIC column (150 mm × 2.1 mm) and guard column (20 mm × 2.1 mm) from Merck Millipore (Germany) and temperature maintained at 35°C. The mobile phase was composed of 10 mM ammonium carbonate and 0.1% ammonium hydroxide in water (solvent A), and acetonitrile (solvent B). The flow rate was set at 100 μL/min with the gradient described previously (Mackay et al., 2015). The mass spectrometer was operated in full MS and polarity switching mode. The acquired spectra were analyzed using XCalibur Qual Browser and XCalibur Quan Browser software 4.2 (Thermo Scientific)
Xcalibur quan browser software 4
Manufactured by Thermo Fisher Scientific
Sourced in United States
XCalibur Quan Browser software 4.2 is a data analysis software tool designed for Thermo Fisher Scientific's mass spectrometry instruments. The software provides a user interface for reviewing and processing quantitative data generated from these instruments.
Automatically generated - may contain errors
Lab products found in correlation
Sequant zic philic column, by Merck Group (2 mentions)
Q exactive hybrid quadrupole orbitrap mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Vanquish uhplc system, by Thermo Fisher Scientific (2 mentions)
Xcalibur qual browser, by Thermo Fisher Scientific (2 mentions)
Isq lt, by Thermo Fisher Scientific (1 mentions)
3 protocols using xcalibur quan browser software 4
1
Metabolite Extraction and LC-MS Analysis of Frozen Heart Tissue
2
Metabolite Extraction and LC-MS Analysis of Frozen Heart Tissue
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Approximately 50 mg of frozen heart was grinded in a mortar submerged in liquid nitrogen. Metabolites were extracted using a solution with 50% methanol, 20% acetonitrile and 30% H2O. LC-MS analysis of the extracted samples was performed using a Q Exactive Hybrid Quadrupole-Orbitrap mass spectrometer coupled to a Vanquish UHPLC system (Thermo Fisher Scientific). The liquid chromatography system was fitted with a Sequant ZIC-pHILIC column (150 mm × 2.1 mm) and guard column (20 mm × 2.1 mm) from Merck Millipore (Germany) and temperature maintained at 35°C. The mobile phase was composed of 10 mM ammonium carbonate and 0.1% ammonium hydroxide in water (solvent A), and acetonitrile (solvent B). The flow rate was set at 100 μL/min with the gradient described previously (Mackay et al., 2015). The mass spectrometer was operated in full MS and polarity switching mode. The acquired spectra were analyzed using XCalibur Qual Browser and XCalibur Quan Browser software 4.2 (Thermo Scientific)
3
Quantification of Gut Short-Chain Fatty Acids
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Cecal content samples (10 mg) were homogenized using Precellys in 0.5 mL of a 5 mM NaOH solution containing internal standards (Acetate-D3, Propionate-D2, Butyrate-13C2 and Valerate-D9). Super-natant (300 µL) was mixed with 500 µL propanol/pyridine solution (3:2 v/v). SCFAs were derivatized using isopropylchloroformate and extracted using 0.5 mL of hexane. Quantification of SCFAs was performed with a gas chromatograph coupled to a mass spectrometer (ISQ LT, ThermoFisher Scientific, Waltham, MA, USA) and XCalibur QuanBrowser software 4.2. (ThermoFisher Scientific).
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