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MB Sample ID: SA184597
Local Sample ID: | B335pos |
Subject ID: | SU002039 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
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 |
---|---|---|---|---|
B335pos | SA184597 | FL022704 | AMPK intestinal KO mice | Genotype |
B335pos | SA184597 | 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. |
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. |
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 |