Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA251610

Local Sample ID:	90
Subject ID:	SU002601
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Male and female

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Sample Preparation:

Sampleprep ID:	SP002607
Sampleprep Summary:	We performed an untargeted metabolomics using a platform, previously described in Contrepois et al., 2015 (Contrepois et al., 2015) to profile plasma extracted metabolites. Samples were thawed and 100 μL of plasma was transferred into a 2mL Protein Lobind Eppendorf tube. In order to precipitate and remove proteins, 400 μL of 1:1:1 methanol:acetonitrile:acetone solution, including 17 internal standards to control for extraction efficiency and evaluate LC-MS performance, was added to each plasma sample while on a cold plate. Samples were placed on the vortex shaker for 15 minutes at 4C then placed at -20C for two hours to allow for sufficient protein precipitation. Samples were centrifuged at 10,000 rpm for 10 min at 4C and the supernatant (metabolite extract) from each sample was carefully removed and placed in a new 2mL Protein Lobind Eppendorf tube. Samples were placed in the turbovap, evaporated to dryness under nitrogen and stored at -80C. Before analysis on the MS, samples were reconstituted in 100 μL 1:1 methanol:water solution and vortexed for 30s, placed in a bath sonicator for 30s and incubated on ice for 30s. This process was repeated 3 times for all samples and were centrifuged at 10,000 rpm for 10 min at 4C. The supernatant of each of these was then transferred to MS tubes and stored at -20C until ran on the mass spectrometer.

Sampleprep ID:

SP002607

Sampleprep Summary:

We performed an untargeted metabolomics using a platform, previously described in Contrepois et al., 2015 (Contrepois et al., 2015) to profile plasma extracted metabolites. Samples were thawed and 100 μL of plasma was transferred into a 2mL Protein Lobind Eppendorf tube. In order to precipitate and remove proteins, 400 μL of 1:1:1 methanol:acetonitrile:acetone solution, including 17 internal standards to control for extraction efficiency and evaluate LC-MS performance, was added to each plasma sample while on a cold plate. Samples were placed on the vortex shaker for 15 minutes at 4C then placed at -20C for two hours to allow for sufficient protein precipitation. Samples were centrifuged at 10,000 rpm for 10 min at 4C and the supernatant (metabolite extract) from each sample was carefully removed and placed in a new 2mL Protein Lobind Eppendorf tube. Samples were placed in the turbovap, evaporated to dryness under nitrogen and stored at -80C. Before analysis on the MS, samples were reconstituted in 100 μL 1:1 methanol:water solution and vortexed for 30s, placed in a bath sonicator for 30s and incubated on ice for 30s. This process was repeated 3 times for all samples and were centrifuged at 10,000 rpm for 10 min at 4C. The supernatant of each of these was then transferred to MS tubes and stored at -20C until ran on the mass spectrometer.