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MB Sample ID: SA254867
| Local Sample ID: | S_189 |
| 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_189 | SA254867 | FL032644 | Without 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 |
