Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA022281

Local Sample ID:	S_153
Subject ID:	SU000461
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Species Group:	Human

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Sample Preparation:

Sampleprep ID:	SP000468
Sampleprep Summary:	A total of 200 study samples were thawed on ice for sample preparation, 400 uL of the thawed serum sample were transferred to labeled tubes on ice where they were mixed with 1200uL of MeOH. Analytical quality control (QC) phenotypic pooled samples (3/Group) were generated by transferring pre-determined volumes of each sample from each respective phenotypic Group’s experimental samples into four different 2.0 mL LoBind tubes. The Phenotypic Pool tubes were vortexed, and 3 aliquots of 400 uL was transferred to Phenotypic Pool tubes for each Group. In addition, a study pool was generated by transferring 200 uL of serum from 25 randomly selected experimental samples into a 10.0 mL tube, vortexed and aliquoted into 10 Study Pool tubes. Methanol was added to all tubes (1200 uL), sample tubes were vortexed for 2 min on a multi-tube vortexer and centrifuged at 16,000 rcf for 5 min. A 1000 µl aliquot of the supernatant was transferred into new pre-labeled 2.0 mL LoBind tubes and lyophilized to complete dryness overnight. Samples were reconstituted with 700 uL of NMR Master Mix solution containing Chenomx ISTD: DSS-d6 and D2O-Phosphate Buffer at 7.4 pH. The tubes were vortexed for 4 min on a multi-tube vortexer and centrifuged at 16,000 rcf for 5 min. A 600 uL of each sample supernatant was transferred into a pre-labeled 5mm 4 NMR tubes for data acquisition on a 700 MHz spectrometer.

Sampleprep ID:

SP000468

Sampleprep Summary:

A total of 200 study samples were thawed on ice for sample preparation, 400 uL of the thawed serum sample were transferred to labeled tubes on ice where they were mixed with 1200uL of MeOH. Analytical quality control (QC) phenotypic pooled samples (3/Group) were generated by transferring pre-determined volumes of each sample from each respective phenotypic Group’s experimental samples into four different 2.0 mL LoBind tubes. The Phenotypic Pool tubes were vortexed, and 3 aliquots of 400 uL was transferred to Phenotypic Pool tubes for each Group. In addition, a study pool was generated by transferring 200 uL of serum from 25 randomly selected experimental samples into a 10.0 mL tube, vortexed and aliquoted into 10 Study Pool tubes. Methanol was added to all tubes (1200 uL), sample tubes were vortexed for 2 min on a multi-tube vortexer and centrifuged at 16,000 rcf for 5 min. A 1000 µl aliquot of the supernatant was transferred into new pre-labeled 2.0 mL LoBind tubes and lyophilized to complete dryness overnight. Samples were reconstituted with 700 uL of NMR Master Mix solution containing Chenomx ISTD: DSS-d6 and D2O-Phosphate Buffer at 7.4 pH. The tubes were vortexed for 4 min on a multi-tube vortexer and centrifuged at 16,000 rcf for 5 min. A 600 uL of each sample supernatant was transferred into a pre-labeled 5mm 4 NMR tubes for data acquisition on a 700 MHz spectrometer.