High performance liquid chromatography-tandem mass spectrometry for the determination of bile acid concentrations in human plasma

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Xiaoqiang XiangMikko Niemi

Abstract

We report a sensitive and robust method to determine cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), lithocholic acid (LCA), ursodeoxycholic acid (UDCA), and their taurine- and glycine-conjugate concentrations in human plasma using liquid chromatography-tandem mass spectrometry. Activated charcoal was utilized to prepare bile acid-free plasma, which served as the biological matrix for the preparation of standard and quality control samples. Plasma sample preparation involved solid-phase extraction. A total of 16 bile acids and 5 internal standards were separated on a reverse column by gradient elution and detected by tandem mass spectrometry in negative ion mode. The calibration curve was linear for all the bile acids over a range of 0.005-5micromol/L. The extraction recoveries for all the analytes fell in the range of 88-101%. Intra-day and inter-day coefficients of variation were all below 10%. A stability test showed that all the bile acids were stable in plasma for at least 6h at room temperature, at least three freeze-thaw cycles, in the -70 degrees C or -20 degrees C freezer for 2 months, and also in the reconstitution solution at 8 degrees C for 48h. Comparison of the matrix effect of bile acid-fr...Continue Reading

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Citations

Jun 10, 2011·European Journal of Clinical Pharmacology·Xiaoqiang XiangMikko Niemi
Nov 12, 2013·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Sai Praneeth R BathenaYazen Alnouti
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Sep 10, 2011·Basic & Clinical Pharmacology & Toxicology·Xiaoqiang XiangMikko Niemi
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Apr 2, 2015·Journal of Pharmacological and Toxicological Methods·Marlies OortsPieter Annaert
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Jun 5, 2016·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Yang YangYongmin Zhang
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