Quantitative analysis of bile acids in human plasma by liquid chromatography-electrospray tandem mass spectrometry: A simple and rapid one-step method

Debora Tagliacozzi, Alessia F. Mozzi, Bruno Casetta, Pierfrancesco Bertucci, Sergio Bernardini, Carmine Di Illio, Andrea Urbani, Giorgio Federici

Research output: Contribution to journalArticlepeer-review


Bile acids play a pivotal role in the metabolism of cholesterol and lipids. Their blood concentrations are important prognostic and diagnostic indicators of hepatobiliary and intestinal dysfunction. This class of molecules comprises a heterogeneous group of compounds with a common cholesterol scaffold. Recently, the introduction of liquid chromatography coupled to tandem mass spectrometry methods has revealed an innovative path in the quantisation of specific bile acids in biological specimens. A robust and sensitive method has been developed based on high performance liquid chromatography separation coupled to an electrospray triple-quadrupole mass spectrometer. Human plasma samples were analysed on a C18 reverse-phase column. The elution profiles were monitored in multiple reaction-monitoring mode, quantifying and identifying each analyte by its own unique precursor to product patterns. A linear correlation over a broad range of bile acid concentrations (0.1-100 μM) was observed. The average recovery period for all of the analysed bile acids was 98±3%. Intra-day and inter-day precision averages were 2% and 5.4%, respectively. The determination was achieved within a single chromatographic run for all unconjugated, glycine- and taurine-conjugated isomeric forms of bile acids. As a proof of principle this method has been validated on a small subset of cholestatic patients (n = 7) and compared to appropriate clinical controls (n = 10). Based upon our encouraging experimental results, the described HPLC separation coupled to tandem mass spectrometry method for the analysis of bile acids in biological samples is deemed a robust and accurate procedure. Consequently, we propose this technique as a suitable candidate method for the identification and quantitation of bile acids in routine analysis.

Original languageEnglish
Pages (from-to)1633-1641
Number of pages9
JournalClinical Chemistry and Laboratory Medicine
Issue number12
Publication statusPublished - 2003


  • Bile acids
  • Hepatobiliary disorders
  • HPLC
  • LC-MS/MS
  • Liquid chromatography-electrospray tandem mass spectrometry
  • Mass spectrometry
  • Routine analysis

ASJC Scopus subject areas

  • Clinical Biochemistry


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