Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA172775

Local Sample ID:	CE003
Subject ID:	SU001928
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Sample Preparation:

Sampleprep ID:	SP001934
Sampleprep Summary:	Accurately weighed freeze-crushed faecal samples were resuspended in 20 times the volume of water (e.g. 12.5 mg of faece in 250 μL water). 250 μL of faecal suspension was homogenized in 747 μL of 0.27 N NaOH by ultrasonication for 1 hr in a screw-cap glass vial containing 3 μL of deuterium-labelled internal standards (d4-CA, d4-GCDCA, d4-TCDCA, d4-CDCA-3S, and d4-LCA, 50 μM each). After incubation for 1 hr at room temperature, pH was adjusted to 8.0 using 8 N HCl, mixed with 110 μL of 0.5 M EDTA/0.5 M TrisHCl. The solution was centrifuged at 15,000 rpm and the supernatant was transferred onto a solid-phase extraction cartridge (Agilent Bond Elut C18, 100 mg/3 mL, preconditioned with 1 mL of methanol and 3 mL of water, three times). The cartridge was washed with 1 mL of water and captured bile acids were eluted with 300 μL of 90% ethanol.

Sampleprep ID:

SP001934

Sampleprep Summary:

Accurately weighed freeze-crushed faecal samples were resuspended in 20 times the volume of water (e.g. 12.5 mg of faece in 250 μL water). 250 μL of faecal suspension was homogenized in 747 μL of 0.27 N NaOH by ultrasonication for 1 hr in a screw-cap glass vial containing 3 μL of deuterium-labelled internal standards (d4-CA, d4-GCDCA, d4-TCDCA, d4-CDCA-3S, and d4-LCA, 50 μM each). After incubation for 1 hr at room temperature, pH was adjusted to 8.0 using 8 N HCl, mixed with 110 μL of 0.5 M EDTA/0.5 M TrisHCl. The solution was centrifuged at 15,000 rpm and the supernatant was transferred onto a solid-phase extraction cartridge (Agilent Bond Elut C18, 100 mg/3 mL, preconditioned with 1 mL of methanol and 3 mL of water, three times). The cartridge was washed with 1 mL of water and captured bile acids were eluted with 300 μL of 90% ethanol.