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MB Sample ID: SA205794
Local Sample ID: | HT1080_ML210_03 |
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_ML210_03 | SA205794 | FL025255 | ML210 | 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 |