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MB Sample ID: SA063434

Local Sample ID:C5
Subject ID:SU001047
Subject Type:Other
Subject Species:Mus musculus
Taxonomy ID:10090

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Combined analysis:

Analysis ID AN001650
Analysis type MS
Chromatography type Reversed phase
Chromatography system Thermo Fisher Orbitrap Exactive plus
Column Phenomenex (Torrance,CA) Hydro-RP C18
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Plus Orbitrap
Ion Mode NEGATIVE
Units ppm

Chromatography:

Chromatography ID:CH001161
Chromatography Summary:Metabolomics profiling was performed with a Dionex Ultimate 3000 quaternary high-performance liquid chromatography (HPLC) pump, column compartment and autosampler coupled Exactive plus Orbitrap mass spectrometer controlled by Xcalibur 2.2 software (Thermo Fisher Scientific, Waltham, MA). LC-MS was run with a modified ion pairing reversed phase negative ion electrospray ionization method (61). A total volume of 10 µL sample is injected and separated on a Phenomenex (Torrance, CA) Hydro-RP C18 column (100 × 2.1 mm, 2.5 µm particle size) using a water/methanol gradient with tributylamine and acetic acid added to the aqueous mobile phase to enhance separation. The HPLC column is maintained at flow rate of 200 µL/min with the temperature of 30 °C. Solvents and gradient are as follows: Solvent A is 3% aqueous methanol with 10 mM tributylamine and 15 mM acetic acid; solvent B is 100% methanol. The gradient is 0 min, 0% B; 5 min, 20% B; 7.5 min, 20% B; 13 min, 55% B; 15.5 min, 95% B; 18.5 min, 95% B; 19 min, 0% B; and 25 min, 0% B. The Exactive plus is operated in negative ion mode at maximum resolving power (140,000), and scanned from m/z 72 -1000 for the first 90 sec and then from m/z 85-1000 for the remainder of the chromatographic run for the detection of small molecule metabolites. The automatic gain control target is 3E6 with a maximum injection time of 100 us, the nitrogen sheath gas flow rate is set at 35, the auxillary gas at 10 and the sweep gas at 1. The capillary voltage is 3.2 kV and both the capillary and heater set at 200 °C, the S-lens was 55.
Instrument Name:Thermo Fisher Orbitrap Exactive plus
Column Name:Phenomenex (Torrance,CA) Hydro-RP C18
Column Temperature:30
Flow Gradient:0 min, 0% B; 5 min, 20% B; 7.5 min, 20% B; 13 min, 55% B; 15.5 min, 95% B; 18.5 min, 95% B; 19 min, 0% B; and 25 min, 0% B.
Flow Rate:200 µL/min
Solvent A:97% water/3% methanol; 15 mM acetic acid; 10 mM tributylamine
Solvent B:100% methanol
Chromatography Type:Reversed phase
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