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MB Sample ID: SA254766
Local Sample ID: | S_85 |
Subject ID: | SU002633 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Female |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002633 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Female |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
S_85 | SA254766 | FL032642 | HDP | Condition |
Collection:
Collection ID: | CO002626 |
Collection Summary: | N/A |
Sample Type: | Blood (serum) |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR002645 |
Treatment Summary: | N/A |
Sample Preparation:
Sampleprep ID: | SP002639 |
Sampleprep Summary: | Samples were stored at -80 °C in the Global Alliance to Prevent Prematurity and Stillbirth (GAPPS) repository before being shipped (on dry ice) to Nutrition Research Institute, UNC-CH, for metabolomics analysis. Serum samples were thawed at 4 °C overnight. Samples were vortexed via a multiple-tube vortex mixer for 2 min at 5000 rpm and then sat on ice for 10 min before sample preparation. Quality control samples (QC pools) were made by pooling 7-µL serum from each parent sample (if sufficient volume is available). An aliquot of the sample (50-µL), including the study sample and QC pool, was transferred into a new pre-labeled tube and mixed with 400-µL methanol containing 500 ng/ml L-tryptophan-d5, vortexed at 5,000 rpm for 2 min, for protein precipitation. All samples were centrifuged at 16, 000 rcf for 10 min at 4°C. The supernatant (350-uL) was dried by a SpeedVac overnight. The dried extracts were reconstituted with 100 µl water-methanol (95:5, v/v) by thoroughly mixing on multiple tube vortex mixer for 10 min at 5000 rpm and centrifuging at 4°C for 10 min at 16,000 rcf. The supernatant (about 100 µL) was transferred to pre-labeled autosampler vials for data acquisition by LC-MS. |
Processing Storage Conditions: | On ice |
Extraction Method: | Vortex with methanol containing 500ng/ml tryptophan-d5 as internal standard |
Extract Storage: | -80℃ |
Sample Resuspension: | Water-Methanol (95:5, v/v) |
Sample Spiking: | Tryptophan-d5 stock solution at 500 ng/mL |
Combined analysis:
Analysis ID | AN004168 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Thermo Vanquish |
Column | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Q Exactive HF-X Orbitrap |
Ion Mode | POSITIVE |
Units | Peak area |
Chromatography:
Chromatography ID: | CH003086 |
Instrument Name: | Thermo Vanquish |
Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
Column Temperature: | 50 |
Flow Gradient: | Time(min) Flow Rate %A %B Curve 1. 0 0.4 99.0 1.0 5 2. 1.00 0.4 99.0 1.0 5 3. 16.00 0.4 1.0 99.0 5 4. 19.00 0.4 1.0 99.0 5 5. 19.50 0.4 99.0 1.0 5 |
Flow Rate: | 0.4 mL/min |
Injection Temperature: | 8 |
Internal Standard: | Tryptophan-d5 |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100% methanol; 0.1% formic acid |
Analytical Time: | 22 |
Weak Wash Solvent Name: | 10% methanol/90% water; 0.1% formic acid |
Strong Wash Solvent Name: | 75% 2-Propanol/25% Water; 0.1% formic acid |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003915 |
Analysis ID: | AN004168 |
Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Xcalibur 4.1.31.9 for data acquisition; Progenesis QI 2.4 for data preprocessing |
Ion Mode: | POSITIVE |
Capillary Temperature: | 275 C |
Collision Energy: | 20-45, ramp |
Fragmentation Method: | CID |
Ion Spray Voltage: | 3.5 kV |
Ionization: | ES+ |
Mass Accuracy: | 5 ppm |
Dataformat: | Profile |
Desolvation Gas Flow: | 45 |
Desolvation Temperature: | 325 C |