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MB Sample ID: SA326875
| Local Sample ID: | BAT_TN_chow_neg_9 |
| Subject ID: | SU003120 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003120 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| BAT_TN_chow_neg_9 | SA326875 | FL038833 | TN | Temperature |
Collection:
| Collection ID: | CO003113 |
| Collection Summary: | Animals were sacrificed immediately by cervical dislocation and tissues were rapidly extracted and immediately snap frozen in liquid nitrogen for metabolite profiling. |
| Sample Type: | Adipose tissue |
Treatment:
| Treatment ID: | TR003129 |
| Treatment Summary: | All mice were housed under a 12 h – 12 h light/dark cycle. Room-temperature mice were housed at 23˚C in ventilated cages with an ACH of 25. Mice housed at thermal neutral conditions were housed in an incubator at 30˚C. Mice exposed to cold were individually housed in an incubator set to 6˚C. Mice were fed a standard diet (Lab Diet 5008) and had free access to food and water. |
Sample Preparation:
| Sampleprep ID: | SP003126 |
| Sampleprep Summary: | For metabolite extraction, tissues were weighed and then homogenized with extraction buffer which consisted of 80% methanol containing Phenylalanine-D₈ internal standard at a 40:1 volume to wet weight ratio. Samples were then centrifuged at 16,000 x g at 4 °C for 15 min. Finally, 50 μL supernatant was transferred to the glass insert for LC-MS detection. |
Combined analysis:
| Analysis ID | AN004937 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Vanquish Horizon |
| Column | Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo orbitrap exploris 240 |
| Ion Mode | NEGATIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH003726 |
| Instrument Name: | Vanquish Horizon |
| Column Name: | Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um) |
| Column Temperature: | 25 °C |
| Flow Gradient: | The linear gradient eluted from 95% B (0.0–1 min), 95% B to 65% B (1–7.0 min), 65% B to 40% B (7.0–8.0 min), 40% B (8.0–9.0 min), 40% B to 95% B (9.0–9.1 min), then stayed at 95% B for 5.9 min. |
| Flow Rate: | 0.4 mL/min |
| Solvent A: | 100% water; 25mM ammonium acetate; 25mM ammonium hydroxide |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004680 |
| Analysis ID: | AN004937 |
| Instrument Name: | Thermo orbitrap exploris 240 |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | ESI source parameters were set as follows: spray voltage, 3500 V or −2800 V, in positive or negative modes, respectively; vaporizer temperature, 350 °C; sheath gas, 50 arb; aux gas, 10 arb; ion transfer tube temperature, 325 °C. The full scan was set as: orbitrap resolution, 60,000; maximum injection time, 100 ms; scan range, 70–1050 Da. The ddMS2 scan was set as: orbitrap resolution, 30,000; maximum injection time, 60 ms; top N setting, 6; isolation width, 1.0 m/z; HCD collision energy (%), 30; Dynamic exclusion mode was set as auto. The metabolites was quantified by Compound Discoverer 3.3. |
| Ion Mode: | NEGATIVE |
