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MB Sample ID: SA301815
| Local Sample ID: | neg-24 |
| Subject ID: | SU002921 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6 |
| Age Or Age Range: | 14-15 weeks old |
| Gender: | Male |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002921 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6 |
| Age Or Age Range: | 14-15 weeks old |
| Gender: | Male |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| neg-24 | SA301815 | FL036620 | 4°C | Temperature |
Collection:
| Collection ID: | CO002914 |
| Collection Summary: | Blood was collected from indwelling arterial catheters and centrifuged for 15 minutes at 10,000 x g, followed by plasma collection. Tissues were harvested from euthanized mice and freeze-clamped in liquid nitrogen. All samples were stored at -80°C until processing. |
| Sample Type: | Multiple |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002930 |
| Treatment Summary: | Mice were individually housed without food or nestlets for 6 hours at room temperature or 4°C. |
Sample Preparation:
| Sampleprep ID: | SP002927 |
| Sampleprep Summary: | For extraction of water-soluble metabolites from plasma samples, 200 µl of methanol was added to 5 µl plasma. This mixture was incubated on ice for 10 minutes, then centrifuged at 17,000g for 10 minutes at 4°C. The supernatant was transferred to an MS vial to run on LCMS. For extraction of hydrophobic metabolites (lipids) from plasma samples, 120 µl of isopropanol kept on dry ice was added to 3 µl plasma. This mixture was vortexed for 10 seconds and kept on dry ice for 10 minutes, then centrifuged at 17,000g for 10 minutes at 4°C. The supernatant was loaded into glass inserts in an MS vial to run on LCMS. For extraction of water-soluble metabolites from tissue samples, frozen tissues were first ground to a powder using a cryomill (Restch, Newtown, PA). Extraction solution kept on ice (40:40:20 methanol:acetonitrile:water) was added to ~10 mg of ground tissue (40 µl solution per mg tissue), vortexed, and kept on ice for 10 minutes, then centrifuged at 17,000g for 10 minutes at 4°C. The supernatant was collected and centrifuged again at the same speed for 10 minutes, and this second supernatant was transferred to an MS vial to run on LCMS. |
Combined analysis:
| Analysis ID | AN004575 | AN004576 | AN004577 | AN004578 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | HILIC | HILIC | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish |
| Column | Waters XBridge BEH Amide (150 x 2.1mm,2.5um) | Waters XBridge BEH Amide (150 x 2.1mm,2.5um) | Agilent InfinityLab Poroshell 120 EC-C8 (150 x 2.1mm,2.7um) | Agilent InfinityLab Poroshell 120 EC-C8 (150 x 2.1mm,2.7um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Plus Orbitrap | Thermo Q Exactive Plus Orbitrap | Thermo Q Exactive Plus Orbitrap | Thermo Q Exactive Plus Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE | POSITIVE | NEGATIVE |
| Units | Intensity | Intensity | Intensity | intensity |
Chromatography:
| Chromatography ID: | CH003439 |
| Chromatography Summary: | Chromatography for hydrophilic compounds |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters XBridge BEH Amide (150 x 2.1mm,2.5um) |
| Column Temperature: | 35 |
| Flow Gradient: | 0 min, 85% B; 2 min, 85% B; 3 min, 80% B; 5 min, 80% B; 6 min, 75% B; 7 min, 75% B; 8 min, 70% B; 9 min, 70% B; 10 min, 50% B; 12 min, 50% B; 13 min, 25% B; 16 min, 25% B; 18 min, 0% B; 23 min, 0% B; 24 min, 85% B; and 30 min, 85% B. |
| Flow Rate: | 150 uL/min |
| Solvent A: | 95% water/5% acetonitrile; 20 mM ammonium acetate; mM ammonium hydroxide, pH 9.45) |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH003440 |
| Chromatography Summary: | Chromatography for lipids |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Agilent InfinityLab Poroshell 120 EC-C8 (150 x 2.1mm,2.7um) |
| Column Temperature: | 35 |
| Flow Gradient: | 0 minutes, 25% B; 2 minutes, 25% B; 4 minutes, 65% B; 16 minutes, 100% B; 20 minutes, 100% B; 21 minutes, 25% B; 27 minutes, 25% B |
| Flow Rate: | 150 uL/min |
| Solvent A: | 90% water/10% methanol; 1mM ammonium acetate ; 0.2% acetic acid |
| Solvent B: | 2% methanol/98% isopropano; 1 mM ammonium acetate; 0.2% acetic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004321 |
| Analysis ID: | AN004575 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Processing performed in El-Maven |
| Ion Mode: | POSITIVE |
| MS ID: | MS004322 |
| Analysis ID: | AN004576 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Processing performed in El-Maven |
| Ion Mode: | NEGATIVE |
| MS ID: | MS004323 |
| Analysis ID: | AN004577 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Processing performed in El-Maven |
| Ion Mode: | POSITIVE |
| MS ID: | MS004324 |
| Analysis ID: | AN004578 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Processing performed in El-Maven |
| Ion Mode: | NEGATIVE |
