Bile acid profiling and quantification in biofluids using ultra-performance liquid chromatography tandem mass...

Area of Science:

• Biochemistry
• Analytical Chemistry
• Metabolomics

Background:

• Bile acids are crucial cholesterol metabolites involved in lipid metabolism and host-microbiota signaling.
• Understanding diverse bile acid functions necessitates advanced, high-throughput analytical techniques.
• Current methods may lack the speed or scope for comprehensive human biofluid analysis.

Purpose of the Study:

• To develop and validate a rapid, high-throughput ultra-performance liquid chromatography (UPLC) method for analyzing bile acids in human biofluids.
• To enable sensitive and quantitative targeted analysis of a wide range of bile acid species.
• To apply the developed method for comparing bile acid profiles in different physiological states.

Main Methods:

• Development of a 15-minute UPLC separation procedure for bile acids from human biofluids with minimal sample preparation.
• Application of high-resolution time-of-flight mass spectrometry (HR-TOF-MS) for broad bile acid profiling.
• Utilization of tandem mass spectrometry (MS/MS) for sensitive, quantitative targeted analysis of 145 bile acid species.

Main Results:

• The UPLC-MS/MS method demonstrated excellent linearity, precision (≈6.5%), and accuracy (81.2-118.9%) for targeted bile acid quantification.
• High recovery rates were achieved for bile acids in serum/plasma (88%) and urine (93%).
• The method successfully elucidated distinct bile acid profiles in preprandial and postprandial states across plasma, serum, and urine samples.

Conclusions:

• A rapid and robust UPLC-MS/MS method was established for comprehensive bile acid analysis in human biofluids.
• This high-throughput platform facilitates detailed investigation of bile acid roles in human physiology and disease.
• The method's application provides insights into dynamic changes in bile acid pools.