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

doi:10.1016/j.plefa.2014.04.005

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

Prostaglandins Leukot Essent Fatty Acids. 2014 Sep;91(3):61-71. doi: 10.1016/j.plefa.2014.04.005. Epub 2014 May 28.

Authors

Nobuaki Tanaka¹, Hiroaki Yamaguchi¹, Ayako Furugen¹, Jiro Ogura¹, Masaki Kobayashi¹, Takehiro Yamada², Nariyasu Mano³, Ken Iseki⁴

Affiliations

¹ Laboratory of Clinical Pharmaceutics & Therapeutics, Division of Pharmasciences, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo 060-0812, Japan.
² Department of Pharmacy, Hokkaido University Hospital, Sapporo 060-8648, Japan.
³ Department of Pharmaceutical Sciences, Tohoku University Hospital, Sendai 980-8574, Japan.
⁴ Laboratory of Clinical Pharmaceutics & Therapeutics, Division of Pharmasciences, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo 060-0812, Japan; Department of Pharmacy, Hokkaido University Hospital, Sapporo 060-8648, Japan. Electronic address: ken-i@pharm.hokudai.ac.jp.

PMID: 24996760
DOI: 10.1016/j.plefa.2014.04.005

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, PGF3α, thromboxane B3 (TXB3), and Δ(17)-6-keto PGF1α) 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.

Keywords: 3-Series prostanoids; Eicosapentaenoic acid; Extracellular measurement; Intracellular measurement; LC/MS/MS.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chromatography, Liquid / methods
Eicosapentaenoic Acid*
Humans
Prostaglandins / analysis*
Spectrometry, Mass, Electrospray Ionization / methods*

Substances

Prostaglandins
Eicosapentaenoic Acid