Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA023629

Local Sample ID:	S_22
Subject ID:	SU000488
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Species Group:	Human

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Sample Preparation:

Sampleprep ID:	SP000495
Sampleprep Summary:	Study Samples and Whole Study Pooled QC Aliquots Preparation: Thawed aqueous saliva samples on ice. Samples were then vortexed and centrifuged at room temperature and at 16,000 rcf for four minutes. A 100 µL aliquot of each experimental sample was transferred to a labeled 2.0 mL Lo-Bind Eppendorf tube to make the study sample aliquots. Whole study pooled QC samples were created by combining 100 µL aliquot from each of the study samples into a 2 mL Lo-Bind eppendorf tube. This QC pooled sample was then vortexed for 30 seconds. Then, ten whole study pooled QC samples of 100 µL each were created. Sample Extraction To all samples (study and whole study pooled QC), 500 µL of Protein Precipitation Buffer with internal standard (0.0125 mg/mL of Tryptophan-d5 in methanol) was added to each tube and vortexed for 4 min at room temperature and at 5,000 rpm. The samples were then centrifuged at room temperature and at 16,000 rcf for 4 min. A 450 µL aliquot of the supernatant from each sample was transferred into pre-labeled 2.0 mL LoBind eppendorf tube and stored at -80 °C for one hour followed by a drying step on a lyophilizer. The residue was reconstituted in 100 µL of reconstitution buffer (95:5 Water:Methanol, v/v) and mixed on a multiple tube vortexer for 10 min at 5,000 rpm. Then, the samples were centrifuged at room temperature for 4 min at 16,000 rcf, and the supernatants were transferred to autosampler vials for data acquisition.

Sampleprep ID:

SP000495

Sampleprep Summary:

Study Samples and Whole Study Pooled QC Aliquots Preparation: Thawed aqueous saliva samples on ice. Samples were then vortexed and centrifuged at room temperature and at 16,000 rcf for four minutes. A 100 µL aliquot of each experimental sample was transferred to a labeled 2.0 mL Lo-Bind Eppendorf tube to make the study sample aliquots. Whole study pooled QC samples were created by combining 100 µL aliquot from each of the study samples into a 2 mL Lo-Bind eppendorf tube. This QC pooled sample was then vortexed for 30 seconds. Then, ten whole study pooled QC samples of 100 µL each were created. Sample Extraction To all samples (study and whole study pooled QC), 500 µL of Protein Precipitation Buffer with internal standard (0.0125 mg/mL of Tryptophan-d5 in methanol) was added to each tube and vortexed for 4 min at room temperature and at 5,000 rpm. The samples were then centrifuged at room temperature and at 16,000 rcf for 4 min. A 450 µL aliquot of the supernatant from each sample was transferred into pre-labeled 2.0 mL LoBind eppendorf tube and stored at -80 °C for one hour followed by a drying step on a lyophilizer. The residue was reconstituted in 100 µL of reconstitution buffer (95:5 Water:Methanol, v/v) and mixed on a multiple tube vortexer for 10 min at 5,000 rpm. Then, the samples were centrifuged at room temperature for 4 min at 16,000 rcf, and the supernatants were transferred to autosampler vials for data acquisition.