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MB Sample ID: SA273733
| Local Sample ID: | A549-GAA-1 |
| Subject ID: | SU002824 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002824 |
| 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 |
|---|---|---|---|---|
| A549-GAA-1 | SA273733 | FL035383 | 10μM | Treatment |
Collection:
| Collection ID: | CO002817 |
| Collection Summary: | Cultured cells were collected by trypsin digestion and low-speed centrifugation. The collected cells were rapidly cooled using liquid nitrogen, and the cooled samples were stored in a cryogenic refrigerator at -80 degrees Celsius. |
| Sample Type: | Lung |
Treatment:
| Treatment ID: | TR002833 |
| Treatment Summary: | A549 cells were seeded in six-well plates and cultured at 37 °C and 5% CO2. When the cell confluence reached about 60%, the corresponding final concentration of GAA or equivalent volume of DMSO was added. The final volume of the medium was 3 ml/well, and the treatment time was 48 hours. |
Sample Preparation:
| Sampleprep ID: | SP002830 |
| Sampleprep Summary: | The collected cells were thawed on ice, then added 500 μM pre-cooled extractant (80% methanol aqueous solution), and whirl for 2 min. Freeze the mixture for 5 min in liquid nitrogen after remove ice for 5 min, it will be whirled for 2 min, circulate this at 3 times. Centrifuge the mixture again with 15000 rpm/min at 4 ℃ for 20 min. Finally take the supernatant into the sample bottle for LC-MS/MS analysis. |
Combined analysis:
| Analysis ID | AN004408 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Nexera UHPLC LC-30A |
| Column | Merck SeQuant ZIC-pHILIC (100 x 2.1mm,5um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | ABI Sciex API 6500 QTrap |
| Ion Mode | POSITIVE |
| Units | relative intensity |
Chromatography:
| Chromatography ID: | CH003308 |
| Instrument Name: | Nexera UHPLC LC-30A |
| Column Name: | Merck SeQuant ZIC-pHILIC (100 x 2.1mm,5um) |
| Column Temperature: | 40℃ |
| Flow Gradient: | 0min A/B (5:95 V/V), 9.5min (50:50 V/V), 11.1 min（5:95 V/V）,14 min (5:95 V/V)) |
| Flow Rate: | 0.4 ml/min |
| Solvent A: | 10mmol/L ammonium acetate+0.3% ammonia |
| Solvent B: | 90% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004156 |
| Analysis ID: | AN004408 |
| Instrument Name: | ABI Sciex API 6500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | The data collection system mainly includes ultra-high performance liquid chromatography and tandem mass spectrometry. LIT and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap mass spectrometer(QTRAP), QTRAP® LC-MS/MS System, equipped with an ESI Turbo Ion-Spray interface, operatingin positive and negative ion mode and controlled by Analyst 1.6.3 software (Sciex). The ESI source operation parameters were as follows: source temperature 450 ∘C; ion spray voltage (IS) 5500 V (positive), -4500 V (negative); ion source gas I (GSI), gas II (GSII), curtain gas (CUR) were set at 40, 55, and 35.0 psi, respectively; the collision gas (CAD) was medium. Instrument tuning and mass calibration were performed with 10 and 100 μmol/L polypropylene glycol solutions in QQQ and LIT modes, respectively. A specific set of MRM transitions were monitored for each period according to the metabolites eluted within this period. |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | Protocol.pdf |
