Analytical Sciences

Measurement of Transport Activities of Bile Acids in Human Multidrug Resistance-Associated Protein 3 Using Liquid Chromatography–Tandem Mass Spectrometry

Kana YAMAGUCHI,* Tsuyoshi MURAI,* Hikaru YABUUCHI,** and Takao KUROSAWA*

*Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Kanazawa, Ishikari-Tobetsu, Hokkaido 061-0293, Japan
**Office of Collaborative Research and Technology Development, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan

A method has been developed for the measurement of transport activities in membrane vesicles obtained from human multidrug resistance-associated protein 3-expressing Sf9 cells for 1β-hydroxy-, 6α-hydroxy- and unsaturated bile acids by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. Calibration curves for the bile acids were linear over the range of 10 to 2000 pmol/mL, and the detection limit was less than 2 pmol/mL for all bile acids using selected reaction monitoring analysis. The method was applied to measurements of adenosine triphosphate-dependent transport activities of the membrane vesicles for the above-mentioned hydroxylated and unsaturated bile acids. The present study demonstrated that the human multidrug resistance-associated protein 3 vesicles accepted 1β-, 6α-hydroxylated and unsaturated bile acids along with common bile acids, such as glycocholic acid and taurolithocholic acid 3-sulfate. The developed method is useful for measurements of bile acid transport activities.

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