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2 protocols using t dca

1

Quantitative Bile Acid Profiling

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Cholic acid (CA), chenodeoxy Cholic acid (CDCA), deoxyCholic acid (DCA), lithoCholic acid (LCA), α muriCholic acid (αMCA), βMCA, ωMCA, ursodeoxyCholic acid (UDCA), hyodeoxyCholic acid (HDCA), murideoxyCholic acid (MDCA), taurine conjugated Cholic acid (T-CA), T-CDCA, T-DCA, T-LCA, T- αMCA, T- βMCA, T- ωMCA, T-UDCA, and T-HDCA were purchased from Steraloids (Newport, Rhode Island). T- ωMCA was given by Dr. Daniel Raftery’s laboratory at the University of Washington Northwest Metabolomics Research Center. For internal standards (ISs) , a total of 5 deuterated (d4) ISs were used. Among them the d4-Cholic acid (d4-CA) was purchased from Toronto Research Chemicals Inc (Ontario, Canada), the d4-deoxyCholic acid (d4-DCA) and d4-chenodeoxyCholic acid (d4-CDCA) were purchased from CDN Isotope Inc (Pointe-Claire, Quebec, Canada), the d4-glycine conjugated CDCA (d4-G-CDCA) was purchased from IsoSciences (Ambler, PA), and d4-lithoCholic acid (d4-LCA) was purchased from Steraloids (Newport, Rhode Island). One mg/mL stock solutions of the individual BAs (for standard curve) and IS were prepared in methanol and water (1:1). The 19 individual BA stock solutions were further diluted in 50% methanol to obtain 10 working standard solutions ranging from 0.05 to 10000 ng/mL). The 5 ISs were mixed to obtain a working IS solution in which the concentration of each IS was 10 µmol/mL.
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2

Comprehensive Bile Acid Quantification

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Deuterated-cholic acid (d4-CA), bile acids (CA, CDCA, LCA, TCA, UDCA, taurine, βMCA, TαMCA, TβMCA, TLCA, TCDCA), and formic acid were purchased from Sigma-Aldrich Chemicals Co. (St Louis, MO, USA). All other bile acid standards (GCDCA, GCA, GDCA, GHDCA, GLCA, GUDCA, HCA, HDCA, ILA, TDCA, THDCA, TUDCA) were purchased from Steraloids Inc. (Newport, RI, USA). Acetonitrile (ACN) and methanol (MeOH) of Optima LC-MS grade quality were purchased from Thermo Fisher Scientific (Auckland, New Zealand).
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