Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA051453

Local Sample ID:	0064.d
Subject ID:	SU000919
Subject Type:	Animal
Subject Species:	Mus musculus
Taxonomy ID:	10090

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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
0064.d	SA051453	FL009916	HFD	Diet
0064.d	SA051453	FL009916	vancomycin	Treatment
0064.d	SA051453	FL009916	plasma	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).

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.

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