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MB Sample ID: SA326886
| Local Sample ID: | EV_24h_C_neg_3 |
| Subject ID: | SU003121 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003121 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| EV_24h_C_neg_3 | SA326886 | FL038834 | Control | Genotype |
| EV_24h_C_neg_3 | SA326886 | FL038834 | Cell | Sample type |
Collection:
| Collection ID: | CO003114 |
| Collection Summary: | For the metabolite extraction from adipocytes, after aspirating the tracing media, the cells were immediately incubated with 500 μL cold methanol containing 1ug/ml internal standard (D8-Phe) for 5 min on dry ice and then scrapped into the Eppendorf tubes. |
| Sample Type: | Adipocytes |
Treatment:
| Treatment ID: | TR003130 |
| Treatment Summary: | Tracing media and corresponding unlabeled media were prepared. Twelve hours before the isotope switch, cell media was replaced with fresh unlabeled media. After switching to the tracing media, the cell samples were collected at 0 h, 1 h, 6 h, 24 h. |
Sample Preparation:
| Sampleprep ID: | SP003127 |
| Sampleprep Summary: | The adipocytes were homogenized in a TissueLyser II (Qiagen) (15 min 30 Hz) at 4 °C. 200 μL of the extracts were mixed with 100 μL Milli-Q water and 200 μL chloroform and centrifuged at 16000g for 5 min at 4 °C. Subsequently, 150 μL of the aqueous solution was centrifugally filtered through a 10-kDa cut-off filter (MRCPRT010, Millipore) to remove proteins. The filtrate was transferred to the glass insert for LC-MS detection. |
Combined analysis:
| Analysis ID | AN004938 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Vanquish Horizon |
| Column | Waters ACQUITY UPLC BEH Amide (100 × 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: | CH003727 |
| Instrument Name: | Vanquish Horizon |
| Column Name: | Waters ACQUITY UPLC BEH Amide (100 × 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 hydroxider |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004681 |
| Analysis ID: | AN004938 |
| 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 |
