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MB Sample ID: SA329866
| Local Sample ID: | POS_06 |
| Subject ID: | SU003152 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Age Or Age Range: | 12 weeks |
| Weight Or Weight Range: | 25 g |
| Gender: | Male |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003152 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Age Or Age Range: | 12 weeks |
| Weight Or Weight Range: | 25 g |
| Gender: | Male |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| POS_06 | SA329866 | FL039067 | 2c44KO | Factor |
Collection:
| Collection ID: | CO003145 |
| Collection Summary: | 100 mg of mouse livers were collected from standard chow diet fed WT and Cyp2c44(-/-) mice. |
| Sample Type: | Liver |
Treatment:
| Treatment ID: | TR003161 |
| Treatment Summary: | Experiments conducted in mice were approved by the Vanderbilt Institutional Animal Care and Use Committee. Male 129/SvJ wild-type (WT) and Cyp2c44(-/-) mice littermates were housed in an Association for Assessment and Accreditation of Laboratory Animal Care–accredited facility. Mice were generated and maintained as described previously (Ghoshal K et al, Diabetes, 2021, doi: 10.2337/db21-0298). Mice were fed SD (~14% fat Purina Laboratory Rodent 5001) up to 12 weeks of age until euthanized to collect livers. |
Sample Preparation:
| Sampleprep ID: | SP003158 |
| Sampleprep Summary: | Mouse liver (100 mg) was homogenized in 500 µl PBS using FisherBrand BeadMill. Homogenates were spiked with 10 µl SPLASH-lipidomics internal standard mix (Avanti Polar Lipids/CRODA), transferred to glass tubes and extracted using the method of Bligh and Dyer. Lipid extracts were evaporated under a gentle stream of nitrogen gas and resuspended in 100 µl of HPLC-grade methanol and chloroform (9:1 ratio). Discovery lipidomics were performed using a Vanquish ultrahigh performance liquid chromatography (UHPLC) system interfaced to a Q Exactive HF quadrupole/orbitrap mass spectrometer (Thermo Fisher Scientific) operating in data-dependent mode. Lipid extracts (5 µl) were injected in both positive and negative ESI modes. Pooled QCs were injected to assess the performance of the LC and MS instruments at the beginning, middle and at the end of each sequence. |
Combined analysis:
| Analysis ID | AN004984 | AN004985 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| Column | Waters ACQUITY UPLC BEH C18 (150 x 2.1mm,1.7um) | Waters ACQUITY UPLC BEH C18 (150 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | peakarea | peakarea |
Chromatography:
| Chromatography ID: | CH003765 |
| Chromatography Summary: | MSRC Lipidomics LC method |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters ACQUITY UPLC BEH C18 (150 x 2.1mm,1.7um) |
| Column Temperature: | 50 |
| Flow Gradient: | Linear gradient from 20% B to 100% B over 7 minutes |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 40% water/60% acetonitrile; 0.1% formic acid; 10 mM Ammonium formate, pH ~3 |
| Solvent B: | 90% isopropanol/10% acetonitrile; 0.1% formic acid; 10 mM Ammonium formate, pH ~3 |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004724 |
| Analysis ID: | AN004984 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS was operating in data-dependent mode (Top7); Raw data data were processed with MS-DIAL version 4.70 in POS-lipidomics mode. |
| Ion Mode: | POSITIVE |
| MS ID: | MS004725 |
| Analysis ID: | AN004985 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS was operating in data-dependent mode (Top7); Raw data data were processed with MS-DIAL version 4.70 in NEG-lipidomics mode. |
| Ion Mode: | NEGATIVE |
