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MB Sample ID: SA251022
Local Sample ID: | IKIDS_SP_18 |
Subject ID: | SU002599 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
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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 |