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MB Sample ID: SA251003
| Local Sample ID: | IKIDS_SP_8 |
| Subject ID: | SU002599 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002599 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| IKIDS_SP_8 | SA251003 | FL032007 | Study Pool | Sample Type |
| IKIDS_SP_8 | SA251003 | FL032007 | 1 | Batch |
Collection:
| Collection ID: | CO002592 |
| Collection Summary: | IKIDS is a prospective birth cohort in Central Illinois. Pregnant women were recruited at two obstetrical clinics in Urbana Champaign IL from 2014-2021. Women were enrolled between 8-14 weeks of pregnancy. Eligibility criteria included 18-40 years of age, English speaking, not in a high risk pregnancy, planning to stay in the area until their child's first birth day and living within 30 minutes of the University of Illinois campus. Health, demographic and life style data and biospecimens were collected at 5 time points across pregnancy and children were followed prospectively from birth including assessments at birth, 1-5 weeks 4.5 months, 7.5 months, 2 years, 3 years, 4 years and 7.5 years of age. Physical and neurobehavioral development were assessed, with a particular focus on the impacts of prenatal chemical exposures (phthalates, phenols, PFAS) or prenatal maternal stress on child health outcomes. |
| Sample Type: | Urine |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002611 |
| Treatment Summary: | NA |
Sample Preparation:
| Sampleprep ID: | SP002605 |
| Sampleprep Summary: | Study samples and study pools (50 µL aliquots) were shipped from the University of Illinois at Urbana-Champaign to the NC HHEAR Hub on dry ice. HHEAR Urine Pool 1 (50 µL each), HHEAR Urine Pool 2 (50 µL each), CHEAR reference urine (50 µL each) and NIST urine (SRM 3672) reference material (50 µL each) were provided by the NC HHEAR Hub. LC-MS grade water (50 µL) was used as blanks. All samples were thawed at 4°C overnight before the preparation. Samples, including study samples, study pool samples, HHEAR reference urine, CHEAR reference urine, NIST reference urine, 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: | 4℃ |
| 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 | AN004111 |
|---|---|
| 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 | m/z |
Chromatography:
| Chromatography ID: | CH003046 |
| Chromatography Summary: | Reverse phase |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| Column Pressure: | 6000-10000 |
| 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: | 8oC |
| 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% 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: | MS003858 |
| Analysis ID: | AN004111 |
| 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 |
| Ion Spray Voltage: | 3.5kV |
| Ionization: | ES+ |
| Mass Accuracy: | 5 ppm |
| Dataformat: | Profile |
| Desolvation Gas Flow: | 55 |
| Desolvation Temperature: | 400 oC |
