Bile Acids Analysis Service | LC–MS/MS Targeted Profiling

Biological Roles of Bile Acids and Why Their Analysis Matters

Bile acids are cholesterol-derived molecules produced in the liver and then reshaped by the gut microbiota. They behave both as detergents that solubilize dietary lipids and as potent signaling molecules that act through receptors such as FXR and TGR5.

On the physicochemical side, bile acids emulsify fats and support absorption of fat-soluble vitamins. On the signaling side, they help coordinate glucose control, lipid handling, inflammatory tone, and energy balance across the liver–gut axis.

Because of these dual roles, bile acid profiles are highly sensitive to changes in liver function, intestinal transit, microbiome composition, and transporter activity. Shifts in primary, secondary, and conjugated bile acids can therefore highlight:

• Cholestatic and other hepatobiliary disorders
• Metabolic diseases and cardiometabolic risk states
• Inflammatory bowel conditions and barrier dysfunction
• Drug-induced liver injury and bile acid transporter inhibition
• Microbiome-driven remodeling of host metabolism

A well-designed bile acids study gives you more than a list of concentrations. It can clarify mechanisms, support target validation, and help identify translational biomarkers that link preclinical models to human disease.

Bile Acids Quantification & Profiling Services at Creative Proteomics

Creative Proteomics provides targeted LC–MS/MS bile acids analysis to support liver, metabolic, microbiome, and drug development projects. The service is built to answer practical questions such as "Which bile acids are changing in my model?", "Are these changes pathway-relevant?", and "Can they serve as biomarkers or safety signals?".

You can select from or combine the following bile acids offerings:

• Targeted bile acids quantification
  Absolute or relative quantitation of primary, secondary, conjugated, muricholic, and phase II bile acids in plasma or serum, bile, feces, tissues, and other matrices.
• Custom bile acids panels
  Panel configuration around your hypothesis, for example FXR/TGR5 signaling, intestinal bile acid pools, transporter substrates, or microbiome-dependent conversions.
• Multi-matrix and multi-species profiling
  Harmonized workflows for human samples and common preclinical species to support cross-species and translational comparisons.
• Integrated statistical and pathway analysis
  Bile acids data packaged with group comparisons, multivariate views, and pathway-level summaries so you can quickly interpret biological meaning.
• Optional multi-omics integration
  Coordinated bile acids profiling with targeted metabolomics, lipidomics, or microbiome sequencing when you need a more complete view of the liver–gut–microbiome axis.

Why Choose Our Bile Acids Analysis

• Broad, isomer-aware panel
  Targeted LC–MS/MS routinely quantifies 60+ primary, secondary, conjugated, muricholic, and phase II bile acids.
• Reproducible quantitative performance
  QC-backed methods with typical CVs around 10–15% for most bile acids and wide, well-characterized calibration ranges.
• One workflow for many matrices and species
  A harmonized method supports plasma/serum, bile, feces, tissues, intestinal contents, urine, and common preclinical models.
• Decision-ready data and easy integration
  You receive concentration tables plus clear group comparisons and core plots (e.g., PCA, volcano), designed to integrate with your metabolomics, lipidomics, and microbiome data.

Bile Acids Analysis Workflow: Step-by-Step Process

Instrumentation and Methods for Bile Acids Profiling

HPLC–HRMS (Agilent 1260 Infinity II + Q Exactive HF-X Orbitrap)

• Reversed-phase C18 LC with negative ESI.
• High-resolution MS/MS for isomer-aware bile acids profiling and confirmation.

UPLC–QTOF (Waters ACQUITY UPLC + Xevo G2-XS QTof)

• Sub-2 μm UPLC gradients for fast, high-resolution separations.
• HRMS and MS/MS for extended bile acids coverage and custom panel development.

GC–MS (Agilent 7890B GC + 5977A)

• Used for derivatized bile acids and pathway intermediates.
• EI-based fragmentation provides complementary structural information to LC–MS.

Agilent 1260 Infinity II HPLC (Figure from Agilent)

Thermo Fisher Q Exactive (Figure from Thermo Fisher)

Waters ACQUITY UPLC System (Figure from Waters)

Agilent 7890B-5977A (Figure from Agilent)

Sample Collection Guidelines for Bile Acids Analysis

*If your available volume or mass is lower than the values above, please contact us to assess feasibility and adjust the protocol.

Demo Results

UPLC–MS/MS MRM chromatogram of a mixed bile acids standard with baseline-separated peaks for CA, DCA, TCA, GCA, TUDCA and β-MCA.

UPLC–MS/MS chromatogram of bile acids in a complex biological sample, showing resolved CA, CCA, DCA, TUDCA and muricholic acids with a low-abundance peak marked S/N > 50.

QC and reproducibility of the bile acids LC–MS/MS method: calibration curves for key bile acids, QC CV% distribution, and PCA scores showing clustered QC injections and separated study groups.

Case Study

A human iPSC-derived hepatocyte screen identifies compounds that inhibit production of Apolipoprotein B

Journal: Communications Biology

Published: 2023

• Background
• Analytical Approach
• Key Findings
• Takeaways for Project Design

A Communications Biology study used human iPSC-derived hepatocytes to screen a focused small-molecule library and identified a family of triazine thiols that suppress secretion of apolipoprotein B (apoB), a key component of atherogenic lipoproteins.

After confirming efficacy in iPSC-hepatocytes and primary human hepatocytes, the team moved into liver-humanized mice to ask a critical mechanistic question:

If VLDL production is inhibited, does the liver redirect cholesterol into bile acids synthesis, and does this perturb the bile acids pool?

Answering this required sensitive, targeted bile acids quantification from limited liver tissue.

Livers from vehicle- and DL-27–treated liver-humanized mice were collected at study end. DL-27 is a soluble triazine thiol analog selected for in vivo testing based on potency and improved formulation properties.

Using our LC–MS metabolomics platform (Creative Proteomics), the authors quantified 12 major human bile acids directly in liver tissue, including: cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid and their glycine- and taurine-conjugated forms. Results were reported on a log scale to capture wide dynamic ranges.

Key features of the bile acids module in this project were:

• Targeted LC–MS/MS panel focused on primary, secondary and conjugated bile acids relevant to human cholesterol catabolism.
• Quantitative comparison between treatment and control groups in a humanized liver model.
• Integration of bile acids data with serum lipids, apoB, Lp(a), ALT and liver histology in the same animals.

The integrated dataset showed that DL-27 treatment in liver-humanized mice:

• Substantially reduced serum apoB, total cholesterol, LDL/VLDL cholesterol, triglycerides and Lp(a), while preserving apoA1 and without increasing ALT or hepatic lipid accumulation.
• Did not cause global disruption of hepatic bile acids; most of the 12 measured bile acids remained unchanged.
• Produced selective increases (up to around one order of magnitude) in several taurine-conjugated bile acids (taurocholic, taurochenodeoxycholic, taurodeoxycholic and taurolithocholic acids) compared with vehicle controls.

Together, these results supported the idea that blocking VLDL export can shift cholesterol toward bile acids synthesis without inducing toxic bile acids accumulation, strengthening the safety and mechanism-of-action narrative for the triazine thiol series.

This case illustrates how a focused bile acids LC–MS/MS panel can:

• Clarify how a candidate therapy reshapes cholesterol handling beyond standard lipid readouts.
• De-risk mechanisms that might otherwise raise concern for cholestasis or bile acids toxicity.
• Add mechanistic depth to complex in vitro–to–in vivo workflows that combine iPSC models, humanized animals and lipid-lowering endpoints.

FAQ of Bile Acid Analysis

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Publications

Here are some of the metabolomics-related papers published by our clients: