Quantification of intracellular and extracellular eicosapentaenoic acid-derived 3-series prostanoids by liquid chromatography/electrospray ionization tandem mass spectrometry

Nobuaki Tanaka, Hiroaki Yamaguchi, Ayako Furugen, Jiro Ogura, Masaki Kobayashi, Takehiro Yamada, Nariyasu Mano, Ken Iseki

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

Abstract

3-Series prostanoids are bioactive lipid mediators synthesized from eicosapentaenoic acid (EPA). Determination of intracellular and extracellular levels of prostanoids is needed to elucidate the mechanism of action, and we therefore developed a method for quantification of intracellular and extracellular levels of 3-series prostanoids (including prostaglandin E3 (PGE3), PGD3, PGF, thromboxane B3 (TXB3), and δ17-6-keto PGF) by using liquid chromatography/electrospray ionization tandem mass spectrometry. The separation of prostanoids was performed with a CAPCELL PAK C18 MG II column (2.0mm×150mm, 3μm) with an isocratic flow of acetonitrile/water/acetic acid (40:60:0.1, v/v/v). This method was validated for measurement of both extracellular and intracellular samples with high levels of precision and accuracy. We applied this method to human lung epithelial A549 cells stimulated with calcium ionophore A23187 under the condition of arachidonic acid or EPA treatment and we could measure PGE3 in both intracellular and extracellular samples.

Original languageEnglish
Pages (from-to)61-71
Number of pages11
JournalProstaglandins Leukotrienes and Essential Fatty Acids
Volume91
Issue number3
DOIs
Publication statusPublished - 2014 Sep

Keywords

  • 3-Series prostanoids
  • Eicosapentaenoic acid
  • Extracellular measurement
  • Intracellular measurement
  • LC/MS/MS

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology

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