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Bile Acids Quantification by Liquid Chromatography-Tandem Mass Spectrometry: Method Validation, Reference Range, and

Elisa Danese1, Davide Negrini2, Mairi Pucci1

  • 1Clinical Biochemistry Section, Department of Neurological, Biomedical and Movement Sciences, University of Verona, 37134 Verona, Italy.

Diagnostics (Basel, Switzerland)
|July 11, 2020
PubMed
Summary
This summary is machine-generated.

A new liquid chromatography-tandem mass spectrometry method accurately quantifies 15 bile acid (BA) species in serum. This validated technique establishes reference ranges and assesses interferences, aiding in diagnosing liver, intestinal, and cardiovascular diseases.

Keywords:
LC–MS/MSbile acidsinterference

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Clinical Diagnostics

Background:

  • Bile acids (BA) are crucial in cholesterol metabolism and serve as biomarkers for hepatobiliary, intestinal, and cardiovascular diseases.
  • Accurate quantification of BA profiles is essential for disease diagnosis and prognosis.
  • Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the established method for BA analysis.

Purpose of the Study:

  • To develop and validate a novel LC-MS/MS method for simultaneous quantification of 15 BA species in human serum.
  • To establish reference ranges for BA concentrations in healthy adults.
  • To evaluate potential in vitro and in vivo interferences affecting BA measurements.

Main Methods:

  • Development and validation of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay.
  • Quantification of 15 bile acid species in serum samples from 130 healthy adults.
  • Assessment of method linearity, accuracy, precision, recovery, and stability.
  • In vitro interference testing with triglycerides, bilirubin, and hemoglobin.
  • In vivo interference evaluation using serum pools from hypercholesterolemic and hyperbilirubinemic patients.

Main Results:

  • The LC-MS/MS method demonstrated excellent linearity (R² > 0.99), accuracy (85-115%), and precision (<10% imprecision).
  • Analyte stability was confirmed for 15 days across various storage conditions.
  • No significant in vitro interference was observed from common biological substances.
  • Distinct BA profiles were identified in hypercholesterolemic and hyperbilirubinemic patient samples compared to healthy controls.

Conclusions:

  • A robust and validated LC-MS/MS method for simultaneous BA quantification in serum has been established.
  • The study provides reference ranges and insights into potential interferences for BA analysis.
  • This method serves as a valuable tool for investigating pathophysiological changes in BA metabolism related to various human diseases.