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MB Sample ID: SA051390
Local Sample ID: | 0015.d |
Subject ID: | SU000919 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000919 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
0015.d | SA051390 | FL009905 | Chow | Diet |
0015.d | SA051390 | FL009905 | none | Treatment |
0015.d | SA051390 | FL009905 | hypothalamus | Tissue |
Collection:
Collection ID: | CO000913 |
Collection Summary: | The mice were fasted for 2 hours and anesthetized with isoflurane before collecting cecum and plasma. |
Sample Type: | Blood |
Blood Serum Or Plasma: | Plasma |
Treatment:
Treatment ID: | TR000933 |
Treatment Summary: | 6 weeks-old male C57BL/6J mice were purchased from Jackson Laboratory and maintained on either a normal chow containing 22% of calories from fat, 23% from protein, and 55% from carbohydrates (Mouse diet 9F 5020; PharmaServ) or a high-fat diet (Open Source Diet, D12492; Research Diets) containing 60% of calories from fat, 20% from protein, and 20% from carbohydrates for 6 weeks. During the last 2 weeks, some of the HFD mice were treated with vancomycin or metronidazole (1 g/L in the drinking water). |
Sample Preparation:
Sampleprep ID: | SP000926 |
Sampleprep Summary: | Plasma: LC-MS samples were prepared from plasma (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 are centrifuged (10 min, 9,000 x g, 4°C), and the supernatants were injected directly. Nucleus accumbens and Hypothalamus: Frozen brain tissue samples were homogenized in 4 volumes of HPLC water (J. T. Baker, Center Valley Pa.) using a TissueLyser II (Qiagen, Hilden, Germany) with 3mm tungsten beads at 20 Hz in two 2-minute cycles. Homogenates were then aliquoted for profiling. Samples were prepared from lysates(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 are centrifuged (10 min, 9,000 x g, 4°C), and the supernatants were injected directly. |
Combined analysis:
Analysis ID | AN001442 |
---|---|
Analysis type | MS |
Chromatography type | HILIC |
Chromatography system | Agilent 1290 infinity II |
Column | Waters Atlantis HILIC (150 x 2mm) |
MS Type | ESI |
MS instrument type | Triple quadrupole |
MS instrument name | Agilent 6495 QQQ |
Ion Mode | POSITIVE |
Units | Peak area |
Chromatography:
Chromatography ID: | CH001013 |
Chromatography Summary: | A 150 x 2.1 mm Atlantis HILIC column (Waters) 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: | Agilent 1290 infinity II |
Column Name: | Waters Atlantis HILIC (150 x 2mm) |
Solvent A: | 100% water; 0.1% formic acid; 10 mM ammonium formate |
Solvent B: | 100% acetonitrile; 0.1% formic acid |
Chromatography Type: | HILIC |
MS:
MS ID: | MS001332 |
Analysis ID: | AN001442 |
Instrument Name: | Agilent 6495 QQQ |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
Ion Mode: | POSITIVE |