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MB Sample ID: SA235996
| Local Sample ID: | PZDu_S_327 |
| Subject ID: | SU002440 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002440 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| PZDu_S_327 | SA235996 | FL029266 | Study Sample | Sample Type |
Collection:
| Collection ID: | CO002433 |
| Collection Summary: | The Atlanta ECHO Cohort is comprised of African American mother-child dyads who reside in the greater metropolitan Atlanta area. Enrollment begins during pregnancy from prenatal care clinics affiliated with two metro hospital systems. During pregnancy, data and biospecimens are obtained during an initial study visit (between 8-14 weeks gestation) and during a second study visit (between 24-30 weeks gestation). In the perinatal period, we collect the residual newborn blood spot (obtained from the neonate during the delivery hospitalization for state metabolic screening). Details of pregnancy complications, birth, and neonatal outcomes are ascertained via medical record abstraction. Children are then followed annually via in-person clinical assessments and maternal questionnaire data. |
| Sample Type: | Blood (serum) |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002452 |
| Treatment Summary: | N/A |
Sample Preparation:
| Sampleprep ID: | SP002446 |
| Sampleprep Summary: | Serum samples (100 µL per tube) were shipped from Emory University to the NC HHEAR Hub on dry ice. The NC HHEAR Hub thawed and transferred 50 µL of the study samples to a new set of tubes and used them for the analysis. An additional 10 µL was taken from the original study sample and transferred to another tube to make the total study pool for this project, and then distributed with 50 µL per aliquot, used as Quality control study pools (QC study pools) throughout the whole analysis. All sample aliquots (50 µL each) and QC study pools (50 µL each) were stored at -80° C until the day for sample preparation. HHEAR reference plasma (50 µL each) and NIST serum (909c) reference material (50 µL each) were provided by the NC HHEAR Hub. The LC-MS grade water (50 µL) was used as blank. Sample preparation was conducted by batch, and all samples (except blank) were thawed at 4°C overnight before the preparation. Samples, including study samples, study pool samples, HHEAR reference plasma, NIST reference serum, 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 |
| Extract Storage: | Described in summary |
Combined analysis:
| Analysis ID | AN003837 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Scientific™ Vanquish™ UPHPLC |
| Column | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,3um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive HF-X Orbitrap |
| Ion Mode | POSITIVE |
| Units | Normalized intensity |
Chromatography:
| Chromatography ID: | CH002841 |
| Instrument Name: | Thermo Scientific™ Vanquish™ UPHPLC |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,3um) |
| 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 |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% methanol; 0.1% formic acid |
| Randomization Order: | Randomized |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003579 |
| Analysis ID: | AN003837 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Instrument: Thermo Q Exactive HFx Software: 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 Flow: | 55 |
| Dry Gas Temp: | 400°C |
| Fragmentation Method: | CID |
| Desolvation Gas Flow: | 55 |
