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MB Sample ID: SA244070

Local Sample ID:MAAP_NIST_3
Subject ID:SU002529
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

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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
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