Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA054762

Local Sample ID:	SM-AADKW
Subject ID:	SU000961
Subject Type:	Human stool
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Sample Preparation:

Sampleprep ID:	SP000968
Sampleprep Summary:	Samples were thawed on ice and then centrifuged (4 ˚C, 5,000 x g) for 5 minutes. Ethanol was evaporated using a gentle stream of nitrogen gas using a nitrogen evaporator (TurboVap LV; Biotage, Charlotte, NC) and stored at -80 ˚C until all samples in the study had been dried. Aqueous homogenates were generated by sonicating each sample in 900 μl of H2O using an ultrasonic probe homogenizer (Branson Sonifier 250) set to a duty cycle of 25% and output control of 2 for 3 minutes. Samples were kept on ice during the homogenization process. The homogenate for each sample was aliquoted into two 10 μL and two 30 μL in 1.5mL centrifuge tubes for LC-MS sample preparation and 30 μL of homogenate from each sample were transferred into a 50 mL conical tube on ice to create a pooled reference sample. The pooled reference mixture was mixed by vortexing and then aliquoted (100 μL per aliquot) into 1.5 mL centrifuge tubes. Aliquots and reference sample aliquots were stored at -80 °C until LC-MS analyses were conducted. Pairs of pooled reference samples were inserted into the queue at intervals of approximately 20 samples in order to assess analytical variance and as a reference to standardize within and across batches by “nearest neighbor” scaling.

Sampleprep ID:

SP000968

Sampleprep Summary:

Samples were thawed on ice and then centrifuged (4 ˚C, 5,000 x g) for 5 minutes. Ethanol was evaporated using a gentle stream of nitrogen gas using a nitrogen evaporator (TurboVap LV; Biotage, Charlotte, NC) and stored at -80 ˚C until all samples in the study had been dried. Aqueous homogenates were generated by sonicating each sample in 900 μl of H2O using an ultrasonic probe homogenizer (Branson Sonifier 250) set to a duty cycle of 25% and output control of 2 for 3 minutes. Samples were kept on ice during the homogenization process. The homogenate for each sample was aliquoted into two 10 μL and two 30 μL in 1.5mL centrifuge tubes for LC-MS sample preparation and 30 μL of homogenate from each sample were transferred into a 50 mL conical tube on ice to create a pooled reference sample. The pooled reference mixture was mixed by vortexing and then aliquoted (100 μL per aliquot) into 1.5 mL centrifuge tubes. Aliquots and reference sample aliquots were stored at -80 °C until LC-MS analyses were conducted. Pairs of pooled reference samples were inserted into the queue at intervals of approximately 20 samples in order to assess analytical variance and as a reference to standardize within and across batches by “nearest neighbor” scaling.