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MB Sample ID: SA184598

Local Sample ID:	B333pos
Subject ID:	SU002039
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

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Subject:

Subject ID:	SU002039
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
B333pos	SA184598	FL022704	AMPK intestinal KO mice	Genotype
B333pos	SA184598	FL022704	HFD	Treatment

Collection:

Collection ID:	CO002032
Collection Summary:	Collect whole blood in a covered test tube,leave the tube in a standing position and wait 30 min.Centrifuge 1500 g 10 min at 4ºC.Take out the serum.
Sample Type:	Blood (serum)

Treatment:

Treatment ID:	TR002051
Treatment Summary:	C57BL/6-AMPKa1flox/flox mice (#014141), and B6.Cg-Tg (Vil-cre) 1000Gum/J (#021504) were purchased from Jackson Laboratory (Bar Harbor, ME). AMPK intestinal KO (AMPK-IKO) mice were generated by cross-breeding C57BL/6-AMPKa1flox/flox mice with B6.Cg-Tg (Vil-cre) 1000Gum/J. All animal procedures were approved by the City of Hope Institutional Animal Care and Use Committee and conducted in accordance with the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals. 6-week-old male mice were housed in a temperature (22-23°C) and light-controlled vivarium with free access to water and high-fat diet (HFD, 60% kcal fat; D12492, Research Diets, New Brunswick, NJ) for 12 weeks.

Treatment ID:

TR002051

Treatment Summary:

C57BL/6-AMPKa1flox/flox mice (#014141), and B6.Cg-Tg (Vil-cre) 1000Gum/J (#021504) were purchased from Jackson Laboratory (Bar Harbor, ME). AMPK intestinal KO (AMPK-IKO) mice were generated by cross-breeding C57BL/6-AMPKa1flox/flox mice with B6.Cg-Tg (Vil-cre) 1000Gum/J. All animal procedures were approved by the City of Hope Institutional Animal Care and Use Committee and conducted in accordance with the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals. 6-week-old male mice were housed in a temperature (22-23°C) and light-controlled vivarium with free access to water and high-fat diet (HFD, 60% kcal fat; D12492, Research Diets, New Brunswick, NJ) for 12 weeks.

Sample Preparation:

Sampleprep ID:	SP002045
Sampleprep Summary:	50 µL of serum was used for metabolites extraction. 200 μL extract solution (acetonitrile: methanol = 1: 1) containing isotopically labeled internal standard mixture was added to each sample. After vortexing for 1 min, the samples were sonicated for 10 min in cold water bath. Then, the samples were incubated at -40 °C for 1 h, and centrifuged at 12000 rpm for 15 min at 4°C. 200 µL of supernatant were transferred to a fresh tube, and dried in a vacuum concentrator at 37 °C. Then, the dried samples were reconstituted in 200 µL of 50% acetonitrile by sonication on ice for 10 min. The constitution was then centrifuged at 13000 rpm for 15 min at 4°C, and 75 µL of supernatant were transferred to a fresh glass vial for LC/MS analysis.

Sampleprep ID:

SP002045

Sampleprep Summary:

50 µL of serum was used for metabolites extraction. 200 μL extract solution (acetonitrile: methanol = 1: 1) containing isotopically labeled internal standard mixture was added to each sample. After vortexing for 1 min, the samples were sonicated for 10 min in cold water bath. Then, the samples were incubated at -40 °C for 1 h, and centrifuged at 12000 rpm for 15 min at 4°C. 200 µL of supernatant were transferred to a fresh tube, and dried in a vacuum concentrator at 37 °C. Then, the dried samples were reconstituted in 200 µL of 50% acetonitrile by sonication on ice for 10 min. The constitution was then centrifuged at 13000 rpm for 15 min at 4°C, and 75 µL of supernatant were transferred to a fresh glass vial for LC/MS analysis.

Combined analysis:

Analysis ID	AN003194
Analysis type	MS
Chromatography type	HILIC
Chromatography system	Agilent 1290 Infinity
Column	Waters Acquity BEH Amide (150 x 2.1mm,1.7um)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	ABI Sciex 5600 TripleTOF
Ion Mode	POSITIVE
Units	Peak area

Chromatography:

Chromatography ID:	CH002362
Instrument Name:	Agilent 1290 Infinity
Column Name:	Waters Acquity BEH Amide (150 x 2.1mm,1.7um)
Chromatography Type:	HILIC

MS:

MS ID:	MS002972
Analysis ID:	AN003194
Instrument Name:	ABI Sciex 5600 TripleTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	UHPLC-MS raw data files were converted into mzXML format using the “msconvert” program from Pro-teoWizard and then analyzed by the XCMS 67 and CAMERA toolbox 68 with R statistical software. The CentWave algorithm in XCMS was used for peak detection. The parameter “peak-width” was set as (5, 20) in units of seconds, referring to the minimum and maximum peak widths for peak detection. The pa-rameter ‘‘snthresh’’ is set as 3 for sensitive peak detection. For multiple UHPLC–MS data files, an or-dered bijective interpolated warping (OBI-Warp) algorithm in XCMS was used for peak alignment.
Ion Mode:	POSITIVE