A liquid chromatography-tandem mass spectrometry-based method for the simultaneous determination of hydroxy sterols and bile acids

John, Clara; Werner, Philipp; Worthmann, Anna; Wegner, Katrin; Tödter, Klaus; Scheja, Ludger; Rohn, Sascha; Heeren, Joerg; Fischer, Markus

doi:10.1016/j.chroma.2014.10.064

Cited by 66 publications

(44 citation statements)

References 33 publications

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“…Serum (10 µL) was added to 10 µL of a 20 ng/µL stock solution of deuterated (d4) glycochenodeoxycholic acid (GCDCA, from Steraloids, Inc.) in methanol, and extracted with ice-cold methanol as previously described 72 . Frozen liver samples (50 mg) or dried fecal pellets (50 mg) were combined with 10 µL of 20 ng/µL d4-GCDCA stock solution and 1.0 mL of ice-cold methanol 72 , and homogenized with 0.5 mm zirconium oxide beads using an air-cooled Bullet Blender (NextAdvance).…”

Section: Methodsmentioning

confidence: 99%

“…Frozen liver samples (50 mg) or dried fecal pellets (50 mg) were combined with 10 µL of 20 ng/µL d4-GCDCA stock solution and 1.0 mL of ice-cold methanol 72 , and homogenized with 0.5 mm zirconium oxide beads using an air-cooled Bullet Blender (NextAdvance). Extracts were dried under vacuum, reconstituted in 2.0 mL of 50% methanol, filtered through 0.2 µm nylon membranes (Pall Life Sciences), and stored at −80 °C.…”

Section: Methodsmentioning

confidence: 99%

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2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-elicited effects on bile acid homeostasis: Alterations in biosynthesis, enterohepatic circulation, and microbial metabolism

Fader

Nault

Zhang

et al. 2017

Sci Rep

113

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2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a persistent environmental contaminant which elicits hepatotoxicity through activation of the aryl hydrocarbon receptor (AhR). Male C57BL/6 mice orally gavaged with TCDD (0.01–30 µg/kg) every 4 days for 28 days exhibited bile duct proliferation and pericholangitis. Mass spectrometry analysis detected a 4.6-fold increase in total hepatic bile acid levels, despite the coordinated repression of genes involved in cholesterol and primary bile acid biosynthesis including Cyp7a1. Specifically, TCDD elicited a >200-fold increase in taurolithocholic acid (TLCA), a potent G protein-coupled bile acid receptor 1 (GPBAR1) agonist associated with bile duct proliferation. Increased levels of microbial bile acid metabolism loci (bsh, baiCD) are consistent with accumulation of TLCA and other secondary bile acids. Fecal bile acids decreased 2.8-fold, suggesting enhanced intestinal reabsorption due to induction of ileal transporters (Slc10a2, Slc51a) and increases in whole gut transit time and intestinal permeability. Moreover, serum bile acids were increased 45.4-fold, consistent with blood-to-hepatocyte transporter repression (Slco1a1, Slc10a1, Slco2b1, Slco1b2, Slco1a4) and hepatocyte-to-blood transporter induction (Abcc4, Abcc3). These results suggest that systemic alterations in enterohepatic circulation, as well as host and microbiota bile acid metabolism, favor bile acid accumulation that contributes to AhR-mediated hepatotoxicity.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-elicited effects on bile acid homeostasis: Alterations in biosynthesis, enterohepatic circulation, and microbial metabolism

Fader

Nault

Zhang

et al. 2017

Sci Rep

113

View full text Add to dashboard Cite

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“…Bile acids pools were measured by liquid chromatography-tandem mass spectrometry (MS/MS) analysis, using chromatographic conditions as described elsewhere (John et al, 2014). The stock solutions of the individual tauro-conjugated and un-conjugated BAs were prepared separately in methanol at a concentration of 1 mg/mL.…”

Section: Methodsmentioning

confidence: 99%

Targeting Bile Acid Receptors: Discovery of a Potent and Selective Farnesoid X Receptor Agonist as a New Lead in the Pharmacological Approach to Liver Diseases

Festa

Marino

Carino³

et al. 2017

Front. Pharmacol.

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Bile acid (BA) receptors represent well-defined targets for the development of novel therapeutic approaches to metabolic and inflammatory diseases. In the present study, we report the generation of novel C-3 modified 6-ethylcholane derivatives. The pharmacological characterization and molecular docking studies for the structure-activity rationalization, allowed the identification of 3β-azido-6α-ethyl-7α-hydroxy-5β-cholan-24-oic acid (compound 2), a potent and selective FXR agonist with a nanomolar potency in transactivation assay and high efficacy in the recruitment of SRC-1 co-activator peptide in Alfa Screen assay. In vitro, compound 2 was completely inactive towards common off-targets such as the nuclear receptors PPARα, PPARγ, LXRα, and LXRβ and the membrane G-coupled BA receptor, GPBAR1. This compound when administered in vivo exerts a robust FXR agonistic activity increasing the liver expression of FXR-target genes including SHP, BSEP, OSTα, and FGF21, while represses the expression of CYP7A1 gene that is negatively regulated by FXR. Collectively these effects result in a significant reshaping of BA pool in mouse. In summary, compound 2 represents a promising candidate for drug development in liver and metabolic disorders.

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“…[15][16][17] Within the past decade, LC-MS has been heavily utilized for the separation and detection of BAs in human and animal model biofluid samples. 9,[18][19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34] The most comprehensive of these methods have a relatively long analytical cycle (> 30 minutes), precluding rapid sample analysis. 21,25,28,30,34 On the other hand, the most rapid methods (< 10 minutes) have limited BA coverage.…”

mentioning

confidence: 99%

“…19,33 The majority of separations have analytical times greater than 20 minutes and intermediate level of BA species coverage (ranging from 11 to 32 species monitored). 9,[18][19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34] Very few LC-MS methods report coverage of tertiary BAs such as sulfate conjugates which have recently been implicated in the important role of the gut microbiota in human metabolism. 9,32,[34][35][36] The analytical foundation of these methods is reversed-phase separation utilizing a C 18 stationary phase, which is suitable for retention and separation of the diverse range of hydrophobicity present among BAs.…”

mentioning

confidence: 99%

Bile Acid Profiling and Quantification in Biofluids Using Ultra-Performance Liquid Chromatography Tandem Mass Spectrometry

et al. 2015

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Bile acids are important end products of cholesterol metabolism. While they have been identified as key factors in lipid emulsification and absorption due to their detergent properties, bile acids have also been shown to act as signaling molecules and intermediates between the host and the gut microbiota. To further the investigation of bile acid functions in humans, an advanced platform for high throughput analysis is essential. Herein, we describe the development and application of a 15 min UPLC procedure for the separation of bile acid species from human biofluid samples requiring minimal sample preparation. High resolution time-of-flight mass spectrometry was applied for profiling applications, elucidating rich bile acid profiles in both normal and disease state plasma. In parallel, a second mode of detection was developed utilizing tandem mass spectrometry for sensitive and quantitative targeted analysis of 145 bile acid (BA) species including primary, secondary, and tertiary bile acids. The latter system was validated by testing the linearity (lower limit of quantification, LLOQ, 0.25?10 nM and upper limit of quantification, ULOQ, 2.5?5 ?M), precision (?6.5%), and accuracy (81.2?118.9%) on inter- and intraday analysis achieving good recovery of bile acids (serum/plasma 88% and urine 93%). The ultra performance liquid chromatography?mass spectrometry (UPLC-MS)/MS targeted method was successfully applied to plasma, serum, and urine samples in order to compare the bile acid pool compositional difference between preprandial and postprandial states, demonstrating the utility of such analysis on human biofluids

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A liquid chromatography-tandem mass spectrometry-based method for the simultaneous determination of hydroxy sterols and bile acids

Cited by 66 publications

References 33 publications

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-elicited effects on bile acid homeostasis: Alterations in biosynthesis, enterohepatic circulation, and microbial metabolism

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-elicited effects on bile acid homeostasis: Alterations in biosynthesis, enterohepatic circulation, and microbial metabolism

Targeting Bile Acid Receptors: Discovery of a Potent and Selective Farnesoid X Receptor Agonist as a New Lead in the Pharmacological Approach to Liver Diseases

Bile Acid Profiling and Quantification in Biofluids Using Ultra-Performance Liquid Chromatography Tandem Mass Spectrometry

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