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MB Sample ID: SA250502
| Local Sample ID: | neg-Pre-TBI-19 |
| Subject ID: | SU002597 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002597 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| neg-Pre-TBI-19 | SA250502 | FL031964 | Control | Treatment |
Collection:
| Collection ID: | CO002590 |
| Collection Summary: | whole blood was collected prior to irradiation (3 days before the first dose of TBI) and at approximately 72h after the end of radiation. |
| Sample Type: | Blood (whole) |
Treatment:
| Treatment ID: | TR002609 |
| Treatment Summary: | 36 patients (28 male and 8 female) received 2 Gy twice a day for three consecutive days (total dose of 12 Gy) at a dose rate 8cGy/min using a six MV medical linear accelerator (Siemens, KD22). The lung was shielded and the dose of supplemental electron fields was no more than 8 Gy. |
Sample Preparation:
| Sampleprep ID: | SP002603 |
| Sampleprep Summary: | GC-MS： 200μL of methanol containing 0.5μg/ml tridecanoic acid (IS) was added into 50μL plasma. After vortexmixing, the solution was centrifuged at 13000 rpm for 15 min at 4 ℃. Then the supernatant was transferred into a new eppendorf tube and dried by vacuum (LABCONCO, USA). The dried sample was re-dissolved in 50μL methoxyamine solution (20 mg/mL) for the oximation reaction (37℃,1.5h), which was followed by a silylation reaction with 50μL MSTFA (37℃,1.5h). Finally, the derivatized sample was centrifuged at 13 000 rpm for 15 min and the supernatant was used for the GC-MS analysis. LC-MS： The internal standards (ISs) were dissolved in acetonitrile including d4-choline (d4-CA, 8μg/mL), C19:0 lysoPC (4μg/mL), C17:0-C17:0 PC (8μg/mL), C17:0-C17:0 PE (8μg/mL), d3-C16:0 free fatty acids (d3-C16:0 FFA, 8μg/mL), d4-cholic acid (CA, 4μg/mL) and d4-chenodeoxycholic acid (d4-CDCA, 4μg/mL). For LC-MS analysis, 200 μL ISs was added into 50 μL plasma sample for the protein precipitation. After vortexing, the sample was centrifuged for 10 min (14,000 rpm, 4 ℃), the supernatant (180 μL ) was redissolved with 100μ L 50% methanol before LC-MS analysis. |
Combined analysis:
| Analysis ID | AN004107 | AN004108 | AN004109 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | GC | Reversed phase | Reversed phase |
| Chromatography system | Agilent 7890A | Thermo TSQ Vantage HPLC-MS/MS | Thermo TSQ Vantage HPLC-MS/MS |
| Column | Agilent DB-5ms (30m x 0.25mm, 0.25um) | Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um) | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | EI | ESI | ESI |
| MS instrument type | Single quadrupole | Triple quadrupole | Triple quadrupole |
| MS instrument name | Agilent 5977 | Thermo TSQ Vantage | Thermo TSQ Vantage |
| Ion Mode | UNSPECIFIED | POSITIVE | NEGATIVE |
| Units | pmoles/l | pmoles/l | pmoles/l |
Chromatography:
| Chromatography ID: | CH003042 |
| Instrument Name: | Agilent 7890A |
| Column Name: | Agilent DB-5ms (30m x 0.25mm, 0.25um) |
| Column Temperature: | 280 |
| Flow Gradient: | The oven temperature was initially set to 70°C, held for 3min, and then increased to 310°C at a rate of 5°C/min, and finally kept for 10min. |
| Flow Rate: | 1.2 mL/min |
| Solvent A: | N/A |
| Solvent B: | N/A |
| Chromatography Type: | GC |
| Chromatography ID: | CH003043 |
| Chromatography Summary: | positive ion mode |
| Instrument Name: | Thermo TSQ Vantage HPLC-MS/MS |
| Column Name: | Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um) |
| Column Temperature: | 50 |
| Flow Gradient: | 0~1 min, 0.1~10% B; 1-5 min, 10-40% B; 5-17 min, 40-100% B; 17-22min, 100% B; 22-22.1 min, 100-10% B |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | water(0.1% formic acid ) |
| Solvent B: | acetonitrile(0.1% formic acid ) |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH003044 |
| Chromatography Summary: | negative ion mode |
| Instrument Name: | Thermo TSQ Vantage HPLC-MS/MS |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 50 |
| Flow Gradient: | 0-1min，0.1%B；1-3min，0.1-40%B；3-5min，40-100%B；5-10.5min，100%B；10.5-10.6min，0.1%B；10.6-18min |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | water(5 mM ammonium bicarbonate ) |
| Solvent B: | 95% methanol/water(5mM ammonium bicarbonate ) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003854 |
| Analysis ID: | AN004107 |
| Instrument Name: | Agilent 5977 |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| MS Comments: | Detector voltage was set at 1.25kV, EI electron impact 70eV. The solvent cutting time was set at 3.5 min. |
| Ion Mode: | UNSPECIFIED |
| MS ID: | MS003855 |
| Analysis ID: | AN004108 |
| Instrument Name: | Thermo TSQ Vantage |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | 350℃capillary temperature, 300℃ vaporize temperature, 40 arbitrary units sheath gas flow rate, 20 arbitrary units auxiliary gas flow rate, 3.0 kV capillary voltage |
| Ion Mode: | POSITIVE |
| MS ID: | MS003856 |
| Analysis ID: | AN004109 |
| Instrument Name: | Thermo TSQ Vantage |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | 350℃capillary temperature, 300℃ vaporize temperature, 40 arbitrary units sheath gas flow rate, 20 arbitrary units auxiliary gas flow rate, -2.5 kV capillary voltage |
| Ion Mode: | NEGATIVE |
