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MB Sample ID: SA054903
Local Sample ID: | SM-A2HYU |
Subject ID: | SU000961 |
Subject Type: | Human stool |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
Analysis ID | AN001513 | AN001514 | AN001515 | AN001516 |
---|---|---|---|---|
Analysis type | MS | MS | MS | MS |
Chromatography type | HILIC | HILIC | Reversed phase | Reversed phase |
Chromatography system | Shimadzu Nexera X2 | Shimadzu Nexera X2 | Shimadzu Nexera X2 | Shimadzu Nexera X2 |
Column | Atlantis HILIC (Waters; Milford,MA) | Luna NH2 (Phenomenex; Torrance,CA) | Waters ACQUITY UPLC BEH C18 | Waters ACQUITY UPLC BEH C8 (1.7um) |
MS Type | ESI | ESI | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Plus Orbitrap | Thermo Q Exactive Orbitrap | Thermo Q Exactive Plus Orbitrap |
Ion Mode | POSITIVE | NEGATIVE | NEGATIVE | POSITIVE |
Units | abundance | abundance | abundance | abundance |
Chromatography:
Chromatography ID: | CH001066 |
Chromatography Summary: | LC-MS samples were prepared from stool homogenates (10 μL) via protein precipitation with the addition of nine volumes of 74.9:24.9:0.2 v/v/v acetonitrile/methanol/formic acid containing stable isotope-labeled internal standards (valine-d8, Isotec; and phenylalanine-d8, Cambridge Isotope Laboratories; Andover, MA). The samples were centrifuged (10 min, 9,000 x g, 4°C), and the supernatants injected directly onto a 150 x 2 mm Atlantis HILIC column (Waters; Milford, MA). The column was eluted isocratically at a flow rate of 250 μL/min with 5% mobile phase A (10 mM ammonium formate and 0.1% formic acid in water) for 1 minute followed by a linear gradient to 40% mobile phase B (acetonitrile with 0.1% formic acid) over 10 minutes. |
Instrument Name: | Shimadzu Nexera X2 |
Column Name: | Atlantis HILIC (Waters; Milford,MA) |
Flow Gradient: | The column was eluted isocratically with 5% A for 1 minute followed by a linear gradient to 40% B over 10 minutes. |
Flow Rate: | 250 µL/min |
Solvent A: | 100% water; 0.1% formic acid; 10 mM ammonium formate |
Solvent B: | 100% acetonitrile; 0.1% formic acid |
Chromatography Type: | HILIC |
Chromatography ID: | CH001067 |
Chromatography Summary: | LC-MS samples were prepared from stool homogenates (30 μL) via protein precipitation with the addition of four volumes of 80% methanol containing inosine-15N4, thymine-d4 and glycocholate-d4 internal standards (Cambridge Isotope Laboratories; Andover, MA). The samples were centrifuged (10 min, 9,000 x g, 4°C) and the supernatants were injected directly onto a 150 x 2.0 mm Luna NH2 column (Phenomenex; Torrance, CA). The column was eluted at a flow rate of 400 μL/min with initial conditions of 10% mobile phase A (20 mM ammonium acetate and 20 mM ammonium hydroxide in water) and 90% mobile phase B (10 mM ammonium hydroxide in 75:25 v/v acetonitrile/methanol) followed by a 10 min linear gradient to 100% mobile phase A. |
Instrument Name: | Shimadzu Nexera X2 |
Column Name: | Luna NH2 (Phenomenex; Torrance,CA) |
Flow Gradient: | The column was eluted with initial conditions of 10% mobile phase A and 90% mobile phase B followed by a 10 min linear gradient to 100% mobile phase A. |
Flow Rate: | 400 µL/min |
Solvent A: | 100% water; 20 mM ammonium acetate; mM ammonium hydroxide |
Solvent B: | 75% acetonitrile/25% methanol; 10 mM ammonium hydroxide |
Chromatography Type: | HILIC |
Chromatography ID: | CH001068 |
Chromatography Summary: | Stool homogenates (30 μL) were extracted using 90 μL of methanol containing PGE2-d4 as an internal standard (Cayman Chemical Co.; Ann Arbor, MI) and centrifuged (10 min, 9,000 x g, 4°C). The supernatants (10 μL) were injected onto a 150 x 2.1 mm ACQUITY BEH C18 column (Waters; Milford, MA). The column was eluted isocratically at a flow rate of 450 μL/min with 20% mobile phase A (0.01% formic acid in water) for 3 minutes followed by a linear gradient to 100% mobile phase B (0.01% acetic acid in acetonitrile) over 12 minutes. |
Instrument Name: | Shimadzu Nexera X2 |
Column Name: | Waters ACQUITY UPLC BEH C18 |
Flow Gradient: | The column was eluted isocratically at a flow rate of 450 µL/min with 20% A for 3 minutes followed by a linear gradient to 100% B over 12 minutes. |
Flow Rate: | 450 µL/min |
Solvent A: | 100% water; 0.01% formic acid |
Solvent B: | 100% acetonitrile; 0.01% acetic acid |
Chromatography Type: | Reversed phase |
Chromatography ID: | CH001069 |
Chromatography Summary: | Lipids (polar and nonpolar) were extracted from stool homogenates (10 μL) using 190 μL of isopropanol containing 1-dodecanoyl-2-tridecanoyl-sn-glycero-3-phosphocholine as an internal standard (Avanti Polar Lipids; Alabaster, AL). After centrifugation (10 min, 9,000 x g, ambient temperature), supernatants (10 μL) were injected directly onto a 100 x 2.1 mm ACQUITY BEH C8 column (1.7 μm; Waters; Milford, MA). The column was eluted at a flow rate of 450 μL/min isocratically for 1 minute at 80% mobile phase A (95:5:0.1 vol/vol/vol 10 mM ammonium acetate/methanol/acetic acid), followed by a linear gradient to 80% mobile-phase B (99.9:0.1 vol/vol methanol/acetic acid) over 2 minutes, a linear gradient to 100% mobile phase B over 7 minutes, and then 3 minutes at 100% mobile-phase B. |
Instrument Name: | Shimadzu Nexera X2 |
Column Name: | Waters ACQUITY UPLC BEH C8 (1.7um) |
Flow Gradient: | The column was eluted isocratically for 1 minute at 80% mobile phase A, followed by a linear gradient to 80% mobile-phase B over 2 minutes, a linear gradient to 100% mobile phase B over 7 minutes, and then 3 minutes at 100% mobile-phase B. |
Flow Rate: | 450 µL/min |
Solvent A: | 95% water/5% methanol; 0.1% acetic acid; 10 mM ammonium acetate |
Solvent B: | 100% methanol; 0.1% acetic acid |
Chromatography Type: | Reversed phase |