A novel fast and simple quantification method for bile acids in human serum by LC-MS/MS

Importance of the topic

Bile acids are steroidal molecules essential for lipid digestion, cholesterol homeostasis and enterohepatic circulation. Altered bile acid profiles serve as biomarkers for liver dysfunction and metabolic diseases. Rapid, sensitive quantification in human serum is therefore critical for clinical research and therapeutic monitoring.

Objectives and Study Overview

This work presents a fast and simple LC-MS/MS method to quantify 27 bile acids simultaneously in human serum. The goals were to streamline sample preparation, shorten analysis time and achieve high sensitivity, repeatability and accurate quantification across physiologically relevant concentrations.

Methodology and Instrumentation

Sample preparation involved protein precipitation of 100 µL serum with 5% ammonium hydroxide in acetonitrile, centrifugation, evaporation under nitrogen and reconstitution in 100 µL solvent. Chromatographic separation was performed on a C8 column (2.7 µm, 2.1×100 mm) using a binary gradient of 0.2% formic acid in water (A) and methanol:isopropanol 1:1 (B). Detection employed negative electrospray ionization and multiple reaction monitoring (MRM) on a Shimadzu Nexera X2 UHPLC coupled to an LCMS-8060 MS. Carryover was minimized by external and needle rinsing protocols.

Key Results and Discussion

• Total run time: 15 min for 27 bile acids with clear isomer resolution.
• Lower limits of quantification (LLOQ) ≤1 ng/mL for 26 compounds; 2.5 ng/mL for 12-dehydrocholic acid; signal-to-noise >10 and RSD <10% at LLOQ.
• Linearity confirmed for 16 bile acids over 1–100 ng/mL with r2 >0.99, accuracy 85–115% and repeatability RSD <5%.

Practical Benefits and Applications

The method’s simplicity and throughput make it suitable for routine metabolic profiling, biomarker discovery and pharmacokinetic studies. Its sensitivity supports detection of low-abundance bile acids, enhancing clinical diagnostics and research in hepatology and metabolic disorders.

Future Trends and Potential Applications

Advances may include integration with automated sample preparation, expansion to additional conjugated species, and coupling with high-resolution MS for structural elucidation. Implementation in large cohort studies could refine bile acid signatures in disease progression and treatment response.

Conclusion

A novel LC-MS/MS workflow delivers rapid, robust quantification of 27 bile acids in serum with minimal preparation. The method meets performance criteria for sensitivity, precision, linearity and accuracy, providing a valuable tool for metabolic and clinical research.

Reference

Jaffuel A., Sambissa S., Huteau A. A novel fast and simple quantification method for bile acids in human serum by LC-MS/MS. MSACL 2017. Shimadzu Corporation.