Bile acid profiling was performed as described.3 (link),12 (link) Livers were homogenized in water (100 mg tissue in 500 μL water), and then, 300 μL of methanol:acetonitrile (v/v, 1:1) was added to a 100 μL aliquot of liver homogenate. All the mixtures were vortexed for 2 minutes and centrifuged at 15,000 rpm for 10 minutes. Two microliters of the supernatants from all samples were injected into the ultraperformance liquid chromatography coupled with an SYNAPT G2-S quadrupole time-of-flight mass spectrometry (QTOFMS, Waters Corporation, Milford, MA). The column type is Acquity UPLC BEH C18 column (2.1 × 100 mm, 1.7 μm). Protocols for the use of a mobile phase gradient and QTOFMS system were reported.12 (link),13 (link) Bile acid species were quantified by measuring their relative abundance as the AUC for each species using standards for comparison.
Serum bile acid quantification was done using the Mouse Total Bile Acids Assay Kit (CrystalChem #80471). Serum samples were diluted 1:5 and were analyzed per kit instructions.
Serum bile acid quantification was done using the Mouse Total Bile Acids Assay Kit (CrystalChem #80471). Serum samples were diluted 1:5 and were analyzed per kit instructions.
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