Return to study ST002115 main page
MB Sample ID: SA205792
| Local Sample ID: | HT1080_ML162_01 |
| Subject ID: | SU002231 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Strain Details: | HT1080 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002231 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Strain Details: | HT1080 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| HT1080_ML162_01 | SA205792 | FL025254 | ML162 | Treatment |
Collection:
| Collection ID: | CO002224 |
| Collection Summary: | Metabolites were extracted from cells in 35 mm culture plates by rapidly aspirating the culture medium and incubating the plates with 0.6 ml of an 80% methanol: 20% water mixture on a cold block on dry ice for 15 min. Next, the cell material was scraped into Eppendorf tubes pre-chilled on ice. After centrifugation at 13,000 RCF for 5 min at 4 °C, the supernatant was collected into a fresh tube and stored on dry ice until analysis. |
| Sample Type: | Cultured cells |
| Storage Conditions: | -80? |
Treatment:
| Treatment ID: | TR002243 |
| Treatment Summary: | Cells were seeded in 35-mm culture plates. When the cell confluence reached 70-80%, cells were treated with RSL3, ML210, or ML162 for 2 hours. |
Sample Preparation:
| Sampleprep ID: | SP002237 |
| Sampleprep Summary: | For analysis by reverse phase chromatography, just before analysis, 500 µL of extract was dried under a nitrogen gas flow and then resuspended in 100 µL of water. For analysis by HILIC chromatography, the extracts were analyzed directly. |
| Processing Storage Conditions: | 4? |
| Extract Storage: | 4? |
Combined analysis:
| Analysis ID | AN003513 | AN003514 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | HILIC |
| Chromatography system | Thermo Accela 1250 | Thermo Accela 1250 |
| Column | Phenomenex Synergi Hydro RP 100 A (100 x 2mm,2.5um) | Waters XBridge BEH Amide (150 x 2.1mm,2.5um,100A) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Exactive Orbitrap | Thermo Exactive Orbitrap |
| Ion Mode | NEGATIVE | NEGATIVE |
| Units | Peak area (top) | Peak area (top) |
Chromatography:
| Chromatography ID: | CH002593 |
| Chromatography Summary: | The gradient was 0 min, 0% B; 2.5 min, 0% B; 5 min, 20% B; 7.5 min, 20% B; 13 min, 55% B; 15.5 min, 95% B; 18.5 min, 95% B; 19 min, 0% B; and 25 min, 0% B. Solvent A was 10 mM tributylamine and 15 mm acetic acid in water; Solvent B was methanol. The injection volume was 10 µL. |
| Instrument Name: | Thermo Accela 1250 |
| Column Name: | Phenomenex Synergi Hydro RP 100 A (100 x 2mm,2.5um) |
| Column Temperature: | 40 |
| Flow Rate: | 200 |
| Solvent A: | 100% water; 15 mM acetic acid; 10 mM tributylamine |
| Solvent B: | 100% methanol |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH002594 |
| Chromatography Summary: | The gradient was 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; 30 min, 85% B. The injection volume was 5 µL. |
| Instrument Name: | Thermo Accela 1250 |
| Column Name: | Waters XBridge BEH Amide (150 x 2.1mm,2.5um,100A) |
| Column Temperature: | 40 |
| Flow Rate: | 150 |
| Solvent A: | 95% water/5% acetonitrile; 20 mM ammonium acetate, pH 9.4 |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003271 |
| Analysis ID: | AN003513 |
| Instrument Name: | Thermo Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The scan range was 80-1000 m/z. Raw data files were converted to mzXML format using msconvert (ProteoWizard). Data was analyzed in the MAVEN software suite and metabolite assignments were made using a previously generated list of retention times derived from pure standard solutions. |
| Ion Mode: | NEGATIVE |
| MS ID: | MS003272 |
| Analysis ID: | AN003514 |
| Instrument Name: | Thermo Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The scan range was 80-1000 m/z. Raw data files were converted to mzXML format using msconvert (ProteoWizard). Data was analyzed in the MAVEN software suite and metabolite assignments were made using a previously generated list of retention times derived from pure standard solutions. |
| Ion Mode: | NEGATIVE |
