Quantification of intracellular and extracellular digoxin and ouabain by liquid chromatography/electrospray ionization tandem mass spectrometry

Hiroaki Yamaguchi, Kazuaki Miyamori, Toshihiro Sato, Jiro Ogura, Masaki Kobayashi, Takehiro Yamada, Nariyasu Mano, Ken Iseki

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

A liquid chromatography/tandem mass spectrometry method for the determination of intracellular accumulation in addition to transcellular transport of digoxin and ouabain in renal epithelial HK-2 cells was developed. The solid-phase extraction Bond Elut® C18 (100mg/1mL) cartridge was used for the extraction of digoxin and ouabain from extracellular (medium) and intracellular (cell lysate) matrices. Chromatographic separation was performed on a CAPCELL PAK C18 MGII column (2.0mm×150mm, 5μm). This method covered a linear range of 0.5-1000ng/mL of concentrations in medium and 0.5-1000ng of concentrations in cell lysate for digoxin and ouabain. The intra-day precision and inter-day precision of analysis were less than 11.9%, and the accuracy was within ±11.6%. The total run time was 16min. Our method was successfully applied to the transport experiments of digoxin and ouabain by HK-2 cell monolayers.

Original languageEnglish
Pages (from-to)73-80
Number of pages8
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume972
DOIs
Publication statusPublished - 2014 Dec 1

Keywords

  • Digoxin
  • Extracellular measurement
  • HK-2 cells
  • Intracellular measurement
  • LC/MS/MS
  • Ouabain

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

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