Measurement of Transport Activities of 3β-Hydroxy-Δ<sup>5</sup>-bile Acids in Bile Salt Export Pump and Multidrug Resistance-Associated Proteins Using LC-MS/MS

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  • Murai Tsuyoshi
    School of Pharmaceutical Sciences, Health Sciences University of Hokkaido
  • Oda Kana
    School of Pharmaceutical Sciences, Health Sciences University of Hokkaido
  • Toyo Terutake
    School of Pharmaceutical Sciences, Health Sciences University of Hokkaido
  • Nittono Hiroshi
    Junshin Clinic Bile Acid Institute
  • Takei Hajime
    Junshin Clinic Bile Acid Institute
  • Muto Akina
    Junshin Clinic Bile Acid Institute
  • Kimura Akihiko
    Department of Pediatrics and Child Health, University School of Medicine
  • Kurosawa Takao
    School of Pharmaceutical Sciences, Health Sciences University of Hokkaido

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  • Measurement of Transport Activities of 3β-Hydroxy-Δ⁵-bile Acids in Bile Salt Export Pump and Multidrug Resistance-Associated Proteins Using LC-MS/MS

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A method has been developed for the measurement of transport activities in membrane vesicles obtained from Sf9 cells for 3β-hydroxy-Δ5-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 1 pmol/mL for 3β-hydroxy-Δ5-bile acids using selected reaction monitoring analysis. The analytical method was applied to measurements of transport activities in membrane vesicles obtained from human multidrug resistance-associated protein 2-, 3-, and human bile salt export pump-expressing Sf9 cells for conjugated 3β-hydroxy-Δ5-bile acids. The present study demonstrated that human multidrug resistance-associated protein 3 vesicles accepted conjugated 3β-hydroxy-Δ5-bile acids along with common bile acids such as glycocholic acid and taurolithocholic acid 3-sulfate.

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