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MB Sample ID: SA366598
| Local Sample ID: | 38 Bottle med |
| Subject ID: | SU003488 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 60 |
| Gender: | Female |
| Cell Strain Details: | BT-474, breast cancer cell line |
| Species Group: | Mammals |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003488 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 60 |
| Gender: | Female |
| Cell Strain Details: | BT-474, breast cancer cell line |
| Species Group: | Mammals |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 38 Bottle med | SA366598 | FL042623 | NA | Sample source |
| 38 Bottle med | SA366598 | FL042623 | Medium | Sample Type |
| 38 Bottle med | SA366598 | FL042623 | NA | Drug Treatment |
Collection:
| Collection ID: | CO003481 |
| Collection Summary: | For medium metabolites, 100 µl of medium was mixed with 400 µl of 100% methanol (80% final). For intracellular metabolite samples, the medium was aspirated, and cells were washed with PBS volume matching the volume of the medium. Metabolites were extracted with ice-cold 80% methanol. The volume of the solvent was 500 µl per 6-cm Petri dish (scaled according to the ratio of surface areas for other cell containers). After adding the methanol solution, cells were scraped from the plates, and all the content was transferred to Eppendorf tubes. |
| Collection Protocol Filename: | DM_metabolomics_samples.txt |
| Sample Type: | Breast cancer cells |
| Collection Method: | 80% methanol extraction |
| Storage Conditions: | -80℃ |
| Collection Vials: | 1.5 ml plastic centrifuge tubes |
| Storage Vials: | 1.5 ml plastic centrifuge tubes |
Treatment:
| Treatment ID: | TR003497 |
| Treatment Summary: | Cells were grown in RPMI-1640 supplemented with 10% dialyzed FBS and treated with various drug concentrations. |
| Treatment Compound: | C75 |
| Treatment Doseduration: | 24 h |
| Treatment Vehicle: | DMSO |
| Cell Growth Container: | 6-cm Petri dishes |
| Cell Media: | RPMI-1640 |
| Cell Envir Cond: | 37C, 5% CO2 |
| Cell Pct Confluence: | ~70% |
| Cell Media Lastchanged: | 24 h before collection |
Sample Preparation:
| Sampleprep ID: | SP003495 |
| Sampleprep Summary: | Intracellular metabolites were extracted with ice-cold 80% methanol, and medium samples were extracted with 100% methanol. The samples were centrifuged at 18,000 g 4C for 20 min. After transferring the supernatant to new tubes, centrifugation was repeated with the same parameters. |
| Sampleprep Protocol Filename: | DM_metabolomics_samples.txt |
| Processing Storage Conditions: | 4℃ |
| Extraction Method: | 80% methanol |
| Extract Enrichment: | None |
| Extract Cleanup: | None |
| Extract Storage: | -80℃ |
| Sample Resuspension: | None |
| Sample Derivatization: | None |
| Sample Spiking: | None |
| Subcellular Location: | Intracellular metabolites and medium metabolites |
Combined analysis:
| Analysis ID | AN005518 | AN005519 |
|---|---|---|
| Chromatography ID | CH004195 | CH004195 |
| MS ID | MS005243 | MS005244 |
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Shimadzu 20AD | Shimadzu 20AD |
| Column | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| MS Type | ESI | ESI |
| MS instrument type | Triple quadrupole | Triple quadrupole |
| MS instrument name | ABI Sciex 5500 QTrap | ABI Sciex 5500 QTrap |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | Counts per second (cps) | Counts per second (cps) |
Chromatography:
| Chromatography ID: | CH004195 |
| Methods Filename: | DM_metabolomics_samples.txt |
| Instrument Name: | Shimadzu 20AD |
| Column Name: | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| Column Pressure: | 900-3000 psi |
| Column Temperature: | 40 |
| Flow Gradient: | 0-12.5 min, 80-30% B; 12.5-15 min, 30% B; 15-15.2 min, 30-80% B; 15.2-22.5 min, 80% B |
| Flow Rate: | 0-20 min, 0.2 ml/min; 20-21 min 0.2-0.3 ml/min; 21-22 min, 0.3 ml/min; 22-22.1 min, 0.2 ml/min; 22.1-22.5 min, 0.2 ml/min |
| Injection Temperature: | 4 |
| Sample Injection: | 1-5 ul |
| Solvent A: | 100% Water; 0.01% ammonium hydroxide; 20 mM ammonium bicarbonate |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS005243 |
| Analysis ID: | AN005518 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | SCIEX Analyst 1.7.3 |
| Ion Mode: | NEGATIVE |
| Capillary Temperature: | 500 °C |
| Capillary Voltage: | -4500 |
| Dry Gas Flow: | 70 |
| Dry Gas Temp: | 500 °C |
| Ion Source Temperature: | 500 °C |
| Ion Spray Voltage: | -4500 |
| Mass Accuracy: | UNIT |
| Source Temperature: | 500 °C |
| Spray Voltage: | -4500 |
| Dataformat: | .WIFF |
| Analysis Protocol File: | DM_metabolomics_samples.txt |
| MS ID: | MS005244 |
| Analysis ID: | AN005519 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | SCIEX Analyst 1.7.3 |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 500 °C |
| Capillary Voltage: | 4500 |
| Dry Gas Flow: | 70 |
| Dry Gas Temp: | 500 °C |
| Ion Source Temperature: | 500 °C |
| Ion Spray Voltage: | 4500 |
| Mass Accuracy: | UNIT |
| Source Temperature: | 500 °C |
| Spray Voltage: | 4500 |
| Dataformat: | .WIFF |
| Analysis Protocol File: | DM_metabolomics_samples.txt |
