Method development and validation of six bile acids for regulated bioanalysis: improving selectivity and sensitivity.

Area of Science:

• Analytical Chemistry
• Biochemistry
• Mass Spectrometry

Background:

• Quantification of bile acids via LC-MS presents challenges for triple quadrupole MS due to lack of primary fragment ions for unconjugated forms.
• Accurate measurement of bile acid profiles is crucial for understanding various physiological and pathological processes.

Purpose of the Study:

• To develop and validate a novel LC-high-resolution/accurate mass MS method for quantifying six key bile acids.
• To overcome limitations of previous MS techniques in bile acid analysis.

Main Methods:

• Development of a liquid chromatography (LC) coupled with high-resolution/accurate mass spectrometry (HR/AM MS) method.
• Utilized target-selected ion monitoring (SIM) mode with dual-period resolution (70,000 and 140,000) for analyte separation.
• Single extraction and single injection procedure for efficiency.

Main Results:

• Successfully developed and validated an LC-HR/AM MS assay for six bile acids: cholic acid, chenodeoxycholic acid, taurocholic acid, deoxycholic acid, lithocholic acid, and ursodeoxycholic acid.
• Achieved robust separation and quantification of target bile acids.
• Demonstrated method's suitability for complex biological matrices.

Conclusions:

• This represents the first fully validated LC-high-resolution/accurate mass assay for quantifying six bile acids in regulated bioanalysis.
• The developed method offers a significant advancement for accurate bile acid quantification.
• The assay provides a reliable tool for clinical and research applications involving bile acid metabolism.