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MB Sample ID: SA246372
| Local Sample ID: | HR1 |
| Subject ID: | SU002551 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002551 |
| 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 |
|---|---|---|---|---|
| HR1 | SA246372 | FL030897 | hypoxia/reoxygenation | Treatment |
Collection:
| Collection ID: | CO002544 |
| Collection Summary: | HUVECs were grown to confluence and subjected to normoxia or hypoxia (2% oxygen for 30 minutes) followed by reoxygenation for 30 minutes. Cells were washed with PBS and scraped from the plates. They were then flash frozen and saved for analysis. |
| Collection Protocol Filename: | HUVECmetabolomics.docx |
| Sample Type: | Endothelial cells |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002563 |
| Treatment Summary: | Cells were subjected to either normoxia or 30 minutes of hypoxia (2%) followed by 30 minutes of reoxygenation. |
Sample Preparation:
| Sampleprep ID: | SP002557 |
| Sampleprep Summary: | Samples were thawed and add with 800μL of 80% methanol, vortexed for 30 s. Then all samples were extracted at 4°C with ultrasound for 30 mins, kept at -40°C for 1 h, and centrifuged at 12000 rpm and 4°C for 15 mins. Finally, supernatant dried down under a nitrogen gas, the residue was resuspended with 100μL of 80% methanol and 5μL of DL-o-Chlorophenylalanine (140μg/mL) for LC-MS analysis. |
Combined analysis:
| Analysis ID | AN004015 | AN004016 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Normal phase | Normal phase |
| Chromatography system | Ultimate 3000LC (Thermo) | Ultimate 3000LC (Thermo) |
| Column | Waters ACQUITY UPLC HSS CN (100 x 2.1mm,1.8um) | Waters ACQUITY UPLC HSS CN (100 x 2.1mm,1.8um) |
| MS Type | ESI | ESI |
| MS instrument type | Single quadrupole | Single quadrupole |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Peak area | Peak area |
Chromatography:
| Chromatography ID: | CH002966 |
| Chromatography Summary: | Separation is performed by Ultimate 3000LC combined with Q Exactive MS (Thermo) and screened with ESI-MS. The LC system is comprised of an ACQUITY UPLC HSS T3 (100×2.1mm 1.8 μm) with Ultimate 3000LC. The mobile phase is composed of solvent A (0.05% formic acid-water) and solvent B (acetonitrile) with a gradient elution (0-1 min, 5% B; 1-12 min, 5-95% B; 12-13.5 min, 95% B; 13.5-13.6 min, 95-5% B; 13.6-16.0 min, 5% B). The flow rate of the mobile phase is 0.3 mL·min-1 . The column temperature is maintained at 40°C, and the sample manager temperature is set at 4°C. Mass spectrometry parameters in ESI+ and ESI- mode are listed as follows: ESI+: Heater Temp 300 °C; Sheath Gas Flow rate, 45 arb; Aux Gas Flow Rate, 15 arb; Sweep Gas Flow Rate, 1 arb; spray voltage, 3.0 kV; Capillary Temp, 350 °C; S-Lens RF Level, 30%. ESI-: Heater Temp 300 °C, Sheath Gas Flow rate, 45 arb; Aux Gas Flow Rate, 15arb; Sweep Gas Flow Rate, 1 arb; spray voltage, 3.2 kV; Capillary Temp,350 °C; S-Lens RF Level, 60%. |
| Instrument Name: | Ultimate 3000LC (Thermo) |
| Column Name: | Waters ACQUITY UPLC HSS CN (100 x 2.1mm,1.8um) |
| Column Temperature: | 40°C |
| Flow Gradient: | 0-1 min, 5% B; 1-12 min, 5-95% B; 12-13.5 min, 95% B; 13.5-13.6 min, 95-5% B; 13.6-16.0 min, 5% B |
| Flow Rate: | 0.3 mL·min-1 |
| Solvent A: | 100% water; 0.05% formic acid |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | Normal phase |
MS:
| MS ID: | MS003762 |
| Analysis ID: | AN004015 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Single quadrupole |
| MS Type: | ESI |
| MS Comments: | ESI+: Heater Temp 300 °C; Sheath Gas Flow rate, 45 arb; Aux Gas Flow Rate, 15 arb; Sweep Gas Flow Rate, 1 arb; spray voltage, 3.0 kV; Capillary Temp, 350 °C; S-Lens RF Level, 30%. |
| Ion Mode: | POSITIVE |
| MS ID: | MS003763 |
| Analysis ID: | AN004016 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Single quadrupole |
| MS Type: | ESI |
| MS Comments: | ESI-: Heater Temp 300 °C, Sheath Gas Flow rate, 45 arb; Aux Gas Flow Rate, 15arb; Sweep Gas Flow Rate, 1 arb; spray voltage, 3.2 kV; Capillary Temp,350 °C; S-Lens RF Level, 60%. |
| Ion Mode: | NEGATIVE |
