LC-MS analysis was performed on a quadrupole-time-of-flight (Q-TOF) 6550 mass spectrometer (Aglient Technologies, Santa Clara, CA) with an electrospray ionization source. The mass spectrometer was interfaced with an Agilent 1290 Infinity II UPLC system (Aglient Technologies, Santa Clara, CA). Metabolites were analyzed in the positive mode over a m/z range of 50–1000 with a C18 T3 reverse-phased column (Waters Corporation, Milford, MA). MS/MS data were generated on the Q-TOF for the identification of perturbed metabolites. The XCMS Online sever (https://xcmsonline.scripps.edu , version 3.5.1) was applied for peak picking, alignment, integration, and extraction of the peak intensities. A two-tailed Welch’s t-test was used for the assessment of metabolite differences between control and treatment groups. The programs of MS-DIAL56 (link) (version 2.70) and MS-FINDER57 (link) (version 2.20) were used for the identification of metabolites.
C18 t3 reverse phased column
Manufactured by Waters Corporation
The C18 T3 reverse-phased column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. The column features a stationary phase consisting of chemically bonded C18 alkyl chains, which provide excellent retention and selectivity for non-polar and moderately polar analytes. The T3 technology enhances the stability and longevity of the column under a variety of mobile phase conditions.
Automatically generated - may contain errors
2 protocols using c18 t3 reverse phased column
1
LC-MS Metabolite Profiling Workflow
2
Fecal Metabolomics Analysis by LC-MS
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The LC–MS
analysis was performed as previously described with minor modifications.58 (link) Briefly, 20 mg fecal sample and 50 mg glass
beads (Sigma-Aldrich, MO) were added to 400 μL cooled methanol
solution (methanol/water, 1:1), followed by homogenization using a
TissueLyser (Qiagen, Hilden, Germany) at 50 Hz for 10 min. The supernatant
was collected after centrifuging for 10 min at 12 000 rpm,
followed by drying up in a speed vacuum (Thermo), and then resuspending
for injection. The mass spectrometer was interfaced with an Agilent
1290 Infinity II UPLC system. The LC–MS analysis was performed
on a quadrupole time-of-flight (Q-TOF) 6530 mass spectrometer (Agilent
Technologies, Santa Clara, CA) with an electrospray ionization source.
Metabolic features were analyzed in the positive mode over a m/z range of 50–1000 with a C18
T3 reverse-phased column (Waters Corporation, Milford, MA). The XCMS
Online sever (https://xcmsonline.scripps.edu , version 3.5.1) was applied for peak picking, alignment, integration,
and extraction of the peak intensities. MS/MS data were generated
on the Q-TOF for the identification of differentiated molecular features.
The software of MS-DIAL (version 2.90)59 (link) and MS-FINDER (version 2.40)60 (link) were used
for the identification of metabolites based on the MS/MS spectrum.
analysis was performed as previously described with minor modifications.58 (link) Briefly, 20 mg fecal sample and 50 mg glass
beads (Sigma-Aldrich, MO) were added to 400 μL cooled methanol
solution (methanol/water, 1:1), followed by homogenization using a
TissueLyser (Qiagen, Hilden, Germany) at 50 Hz for 10 min. The supernatant
was collected after centrifuging for 10 min at 12 000 rpm,
followed by drying up in a speed vacuum (Thermo), and then resuspending
for injection. The mass spectrometer was interfaced with an Agilent
1290 Infinity II UPLC system. The LC–MS analysis was performed
on a quadrupole time-of-flight (Q-TOF) 6530 mass spectrometer (Agilent
Technologies, Santa Clara, CA) with an electrospray ionization source.
Metabolic features were analyzed in the positive mode over a m/z range of 50–1000 with a C18
T3 reverse-phased column (Waters Corporation, Milford, MA). The XCMS
Online sever (
and extraction of the peak intensities. MS/MS data were generated
on the Q-TOF for the identification of differentiated molecular features.
The software of MS-DIAL (version 2.90)59 (link) and MS-FINDER (version 2.40)60 (link) were used
for the identification of metabolites based on the MS/MS spectrum.
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