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MB Sample ID: SA354224
| Local Sample ID: | Les-Jas-20230207-08 |
| Subject ID: | SU003382 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003382 |
| 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 |
|---|---|---|---|---|
| Les-Jas-20230207-08 | SA354224 | FL041330 | Brain | Sample source |
| Les-Jas-20230207-08 | SA354224 | FL041330 | 4hrs | Treatment |
Collection:
| Collection ID: | CO003375 |
| Collection Summary: | GBM cells were cultured under the conditions indicated in the manuscript, then metabolites were collected using 80% methanol extraction. |
| Sample Type: | Brain |
Treatment:
| Treatment ID: | TR003391 |
| Treatment Summary: | metabolites isolated from Cki treated cells. Samples 1-3 are control treatment, Sample 4-6 are 1 hour CKi treatment, Samples 7-9 are 4 hours CKi treatmet, and 10-12 are 8 hours CKi treatment. |
Sample Preparation:
| Sampleprep ID: | SP003389 |
| Sampleprep Summary: | Isolated samples were dried using a SpeedVac. Acetonitrile (50%) was added to the tube for reconstitution following overtaxing for 30s. Sample solution was then centrifuged for 15min at 20,000g and 4°C. Supernatant was collected for LC-MS analysis. |
Combined analysis:
| Analysis ID | AN005346 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 |
| Column | Water's Xbridge amide (100 x 3mm, 3.5 um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Plus Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH004048 |
| Chromatography Summary: | Samples were analyzed by high-performance LC (HPLC) and high-resolution MS and MS/MS (HPLC-MS/MS). The system consists of Thermo Q Exactive with an electrospray source and an UltiMate3000 (Thermo Fisher Scientific) series HPLC consisting of a binary pump, degasser, and autosampler outfitted with an XBridge Amide column (Waters; dimensions of 4.6 mm by 100 mm and a 3.5-μm particle size). The mobile phase A contained 95% water/5% acetonitrile (v/v), 20 mM ammonium hydroxide, and 20 mM ammonium acetate (pH 9.0); phase B was 100% acetonitrile. The gradient was performed as follows: 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1 to 18 min, 75% A; and 18 to 25 min, 15% A with a flow rate of 400 µl/min. |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Water's Xbridge amide (100 x 3mm, 3.5 um) |
| Column Temperature: | - |
| Flow Gradient: | 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1 to 18 min, 75% A; and 18 to 25 min, 15% A |
| Flow Rate: | 400 μl/min |
| Solvent A: | 95% water/5% acetonitrile; 20 mM ammonium hydroxide; 20 mM ammonium acetate (pH 9.0) |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS005076 |
| Analysis ID: | AN005346 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The capillary of the electrospray ionization source was set to 275°C, with sheath gas at 45 arbitrary units, auxiliary gas at 5 arbitrary units, and the spray voltage at 4.0 kV. In positive/negative polarity switching mode, a mass/charge ratio (m/z) scan range from 70 to 850 was chosen and MS1 data were collected at a resolution of 70,000. The automatic gain control target was set at 1 × 106, and the maximum injection time was 200 ms. The top five precursor ions were subsequently fragmented, in a data-dependent manner, using the higher-energy collisional dissociation cell set to 30% normalized collision energy in MS2 at a resolution power of 17,500. |
| Ion Mode: | UNSPECIFIED |
