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MB Sample ID: SA177699
Local Sample ID: | SP_1 |
Subject ID: | SU001992 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male and female |
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Sample Preparation:
Sampleprep ID: | SP001998 |
Sampleprep Summary: | Ninety-two fecal samples (250-300 mg) were randomized and homogenized in 50:50 acetonitrile:water (5 mL/ mg fecal mass) in MagNALyser tubes with ceramic beads, using an Omni Bead Ruptor (5 meters per second, two 30-sec cycles, 15 sec dwell time in between cycles). Samples were centrifuged at 16,000 relative centrifugal force (rcf) for 15 min, the supernatant was transferred, and centrifuged again at 16,000 rcf for 5 min. Quality control (QC) samples were prepared by pooling 70 µL supernatant from each of the study samples and processed identically to the study samples. An aliquot of the supernatant (100 mL) from each sample was transferred to 2.0 mL tubes and dried overnight by SpeedVac and then reconstituted in 200 mL of reconstitution solution (95:5 water-methanol solvent containing 500 ng/ml tryptophan d-5). Samples were centrifuged at 16,000 rcf at 4°C for 4 minutes and the supernatants were transferred to autosampler vials. The study samples were randomized with interspersed QC pools before data acquisition. An injection volume of 5 mL was used for the UPLC-MS analysis. |
Sampleprep Protocol Filename: | OA-Fecal LCMS procedures |
Processing Method: | Extraction |
Processing Storage Conditions: | On ice |
Extract Storage: | -80℃ |
Sample Resuspension: | 95:5 water-methanol solvent containing 500 ng/ml tryptophan d-5 |