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MB Sample ID: SA244085
| Local Sample ID: | MAAP_S_30 |
| Subject ID: | SU002529 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002529 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| MAAP_S_30 | SA244085 | FL030600 | Study Sample | Sample Type |
Collection:
| Collection ID: | CO002522 |
| Collection Summary: | In 2014, the Microbes, Allergy, Asthma and Pets (MAAP) study (P01 AI089473) was launched. All women ages 18-49 years scheduled for prenatal visits at Henry Ford Health, expected to deliver during the recruitment period at a Henry Ford Health hospital, and whose residence was in a predefined area of Detroit and metropolitan Detroit suburbs were potentially eligible. There were no medical insurance requirements. Between 2014 and 2016, 141 women were enrolled; their infant was followed through age 18 months (with continuing follow-up through early/mid childhood underway through the NIH ECHO program under the CREW birth cohort consortium (5UG3OD023282, Gern PI). Blood and stool samples were obtained from the mother prenatally and stool specimens were obtained from the infant at 5 time points (1-week, 1-, 3-, 6- and 18-months) with 16S sequencing already completed. Blood was also obtained from the infant at pre-defined intervals. Please contact Andrea Cassidy-Bushrow at acassid1@hfhs.org for questions related to the subject characteristics and outcomes. |
| Sample Type: | Blood (plasma) |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002541 |
| Treatment Summary: | N/A |
Sample Preparation:
| Sampleprep ID: | SP002535 |
| Sampleprep Summary: | Study samples (500-1000 µL aliquots) were shipped from the Henry Ford Health System to the NC HHEAR Hub on dry ice. The NC HHEAR Hub thawed and aliquoted the study samples at 50 µL per aliquot. An additional 10 µL was taken from each study sample, mixed, and distributed at 50 µL per aliquot, which were used as QC study pools throughout the study. HHEAR reference plasma (50 µL each) and NIST plasma (1950) reference material (50 µL each) were provided by the NC HHEAR Hub. LC-MS grade water (50 µL) was used as blank. All samples were thawed at 4°C overnight before the preparation. Samples, including study samples, study pool samples, HHEAR reference plasma, NIST reference plasma, and blanks were mixed with 400 µL methanol containing 500 ng/mL tryptophan-d5 as internal standard and vortexed by a multiple tube vortex mixer for 2 min at 5000 rpm at room temperature. All samples were centrifuged at 16,000 rcf for 10 min at 4°C. The supernatant (350 µL) was transferred into a pre-labeled 2.0 mL Lo-bind Eppendorf tube, dried by a SpeedVac overnight, and stored at -80° C. For immediate analysis, 100 µL of water-methanol solution (95:5, v/v) was used to reconstitute the dried extracts. Samples were thoroughly mixed on a multiple tube vortex mixer for 10 min at 5000 rpm at room temperature and then centrifuged at 4°C for 10 min at 16,000 rcf. The supernatant 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 | AN003977 |
|---|---|
| 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: | CH002940 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| Column Pressure: | 6000-10000 psi |
| 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 6. 22.00 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 min |
| Weak Wash Solvent Name: | 10:90 Methanol:Water with 0.1% FA solution |
| Strong Wash Solvent Name: | 75:25 2-Propanol:Water with 0.1% FA solution |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003711 |
| Analysis ID: | AN003977 |
| 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: | 320 °C |
| Capillary Voltage: | 3.5 kV |
| Collision Energy: | 20-45, ramp |
| Collision Gas: | N2 |
| Dry Gas Temp: | 400°C |
| Fragmentation Method: | CID |
| Desolvation Gas Flow: | 55 |
