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MB Sample ID: SA307256
| Local Sample ID: | SUB11589p2_Spl31 |
| Subject ID: | SU002945 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002945 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| SUB11589p2_Spl31 | SA307256 | FL036991 | ICC21 | cell line |
| SUB11589p2_Spl31 | SA307256 | FL036991 | Infigratinib | treatment |
| SUB11589p2_Spl31 | SA307256 | FL036991 | N/A | labeling status |
Collection:
| Collection ID: | CO002938 |
| Collection Summary: | 1 million ICC cells were seeded in 6 cm dishes under indicated treatments with triplicates to identify metabolic characteristics. Fresh media was provided 2 hours before harvest. For metabolite collection, media was completely aspirated, and cells were washed with ice-cold saline quickly. After washing, fully remove saline and cells can be scrapped in 1 ml pre-cooled methanol (-20°C) with internal standards (Cambridge Isotope Laboratories, MSK-A2-1.2), and transferred to glass vials, stored at -80°C until extraction. |
| Sample Type: | Tumor cells |
Treatment:
| Treatment ID: | TR002954 |
| Treatment Summary: | ICC13-7 cells were treated with DMSO or 100 nM infigratinib for 24 hours, then labeled with 13C6-Glucose for indicated time points (0,1,24 hours). ICC21 cells were treated with DMSO, 100 nM infigratinib, 100 nM afatinib or combo for 24 hours. |
Sample Preparation:
| Sampleprep ID: | SP002951 |
| Sampleprep Summary: | Samples were dried under N2 flow. Samples were resuspended in acetonitrile 50% in water. The volume was scaled to the biomass. 15ul was used for the lowest biomass, and all other were scaled accordingly. Standard mixes were prepared at 100uM and run after the samples to allow for identification of the targets |
Combined analysis:
| Analysis ID | AN004631 | AN004632 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| Column | Thermo BETASIL Diol (150 x 2.1mm,5um) | Thermo BETASIL Diol (150 x 2.1mm,5um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Orbitrap ID-X tribrid | Thermo Orbitrap ID-X tribrid |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | area analyzed by Compound discoverer (counts x seconds) | area analyzed by Compound discoverer (counts x seconds) |
Chromatography:
| Chromatography ID: | CH003485 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Thermo BETASIL Diol (150 x 2.1mm,5um) |
| Column Temperature: | 40 |
| Flow Gradient: | The LC program was as follow: starting at 93% B, to 40% B in 19 min, then to 0% B in 9 min, maintained at 0% B for 5 min, then back to 93% B in 3 min and re-equilibrated at 93% B for 9 min. |
| Flow Rate: | The flow rate was maintained at 0.15 mL/min, except for the first 30 seconds where the flow rate was uniformly ramped from 0.05 to 0.15 mL/ min. |
| Solvent A: | 20 mMAmmonium Carbonate, 0.1% Ammonium hydroxide, in Water |
| Solvent B: | Acetonitrile 97%, in water |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004378 |
| Analysis ID: | AN004631 |
| Instrument Name: | Thermo Orbitrap ID-X tribrid |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Data was acquired on the ID-X in switching polarities at 120000 resolution, with an AGC target of 1e5, and a m/z range of 65 to 1000. MS1 data is acquired in switching polarities for all samples. Data is analyzed in Compound Discoverer 3.3 (CD, ThermoFisher Scientific). A mix of standard of each target was run along the samples and used as the unlabeled reference for the CD labelling workflow. Isotopomer distribution is found in the exchange column, as % of 0 labelled carbon, 1 labelled carbon, 2 labelled carbon, etc. Those are corrected for natural abundance. For the compounds that couldn’t be analyzed by CD, the areas have been extracted with Tracefinder manually. |
| Ion Mode: | POSITIVE |
| MS ID: | MS004379 |
| Analysis ID: | AN004632 |
| Instrument Name: | Thermo Orbitrap ID-X tribrid |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Data was acquired on the ID-X in switching polarities at 120000 resolution, with an AGC target of 1e5, and a m/z range of 65 to 1000. Data is analyzed in Compound Discoverer 3.3 (CD, ThermoFisher Scientific). A mix of standard of each target was run along the samples and used as the unlabeled reference for the CD labelling workflow. Isotopomer distribution is found in the exchange column, as % of 0 labelled carbon, 1 labelled carbon, 2 labelled carbon, etc. Those are corrected for natural abundance. For the compounds that couldn’t be analyzed by CD, the areas have been extracted with Tracefinder manually. |
| Ion Mode: | NEGATIVE |
