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MB Sample ID: SA212614
| Local Sample ID: | ML-08_Positive |
| Subject ID: | SU002317 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Genotype Strain: | HOG |
| Gender: | Male |
| Cell Strain Details: | Human oligodendroglioma cell line |
| Cell Counts: | 500,000 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002317 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Genotype Strain: | HOG |
| Gender: | Male |
| Cell Strain Details: | Human oligodendroglioma cell line |
| Cell Counts: | 500,000 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| ML-08_Positive | SA212614 | FL025938 | HOG R132H | Genotype |
| ML-08_Positive | SA212614 | FL025938 | BAY2402234 | Treatment |
Collection:
| Collection ID: | CO002310 |
| Collection Summary: | Cells were washed with ice-cold saline and snap frozen in liquid nitrogen. |
| Sample Type: | Cultured cells |
Treatment:
| Treatment ID: | TR002329 |
| Treatment Summary: | HOG cells were plated in 6-well plates (0.5 × 10^6 cells per well). 24 hours later, HOG-EV or HOG-R132H cells were treated for 24 hours with 10 nM BAY 2402234 or DMSO. |
Sample Preparation:
| Sampleprep ID: | SP002323 |
| Sampleprep Summary: | Metabolites were extracted in 80% methanol, vortexed for 20 min at 4°C, and centrifuged (21,100 × g for 10 min at 4°C). Metabolite samples were dried using a CentriVap (Labconco) or SpeedVac (Thermo Fisher) concentrator. Dried metabolites were resuspended in 40-50 µL 80% acetonitrile, vortexed for 20 min at 4°C, and centrifuged (21,100 × g for 10 min at 4°C). 10 µL was injected and analyzed with a Q-Exactive HF-X hybrid quadrupole-orbitrap mass spectrometer (Thermo Fisher) coupled to a Vanquish Flex UHPLC system (Thermo Fisher). |
Combined analysis:
| Analysis ID | AN003640 | AN003641 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Vanquish Flex UHPLC | Vanquish Flex UHPLC |
| Column | Millipore ZIC-pHILIC | Millipore ZIC-pHILIC |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF-X Orbitrap | Thermo Q Exactive HF-X Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | TIC-Corrected Peak Area | TIC-Corrected Peak Area |
Chromatography:
| Chromatography ID: | CH002695 |
| Chromatography Summary: | Chromatographic resolution of metabolites was achieved using on a Millipore ZIC-pHILIC column using a linear gradient of 10 mM ammonium formate pH 9.8 and acetonitrile. |
| Instrument Name: | Vanquish Flex UHPLC |
| Column Name: | Millipore ZIC-pHILIC |
| Flow Gradient: | linear gradient of 10 mM ammonium formate pH 9.8 and acetonitrile |
| Solvent A: | 100% water; 10 mM ammonium formate pH 9.8 |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003391 |
| Analysis ID: | AN003640 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Spectra were acquired with a resolving power of 60,000 full width at half maximum (FWHM), a scan range set to 80–1,200 m/z, and polarity switching. Data-dependent MS/MS data was acquired on unlabeled pooled samples to confirm metabolite IDs when necessary. Peaks were integrated using El-Maven 0.12.0 software (Elucidata) or TraceFinder 5.1 SP2 software (Thermo Fisher). Total ion counts were quantified using Freestyle 1.7 SP1 software (Thermo Fisher). Peaks were normalized to total ion counts using the R statistical programming language. |
| Ion Mode: | POSITIVE |
| MS ID: | MS003392 |
| Analysis ID: | AN003641 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Spectra were acquired with a resolving power of 60,000 full width at half maximum (FWHM), a scan range set to 80–1,200 m/z, and polarity switching. Data-dependent MS/MS data was acquired on unlabeled pooled samples to confirm metabolite IDs when necessary. Peaks were integrated using El-Maven 0.12.0 software (Elucidata) or TraceFinder 5.1 SP2 software (Thermo Fisher). Total ion counts were quantified using Freestyle 1.7 SP1 software (Thermo Fisher). Peaks were normalized to total ion counts using the R statistical programming language. |
| Ion Mode: | NEGATIVE |
