Quantification of intracellular and extracellular prostanoids stimulated by A23187 by liquid chromatography/electrospray ionization tandem mass spectrometry

Ayako Furugen, Hiroaki Yamaguchi, Nobuaki Tanaka, Hajime Ito, Kazuaki Miyamori, Asuka Fujikawa, Natsuko Takahashi, Jiro Ogura, Masaki Kobayashi, Takehiro Yamada, Nariyasu Mano, Ken Iseki

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

Prostanoids are bioactive substances that contribute to various biological and pathological processes. To evaluate both extracellular and intracellular levels of prostanoids at the same time, we developed methods for quantification of extracellular and intracellular levels of prostanoids, including prostaglandin E 2 (PGE 2), PGD 2, PGF , 6-keto PGF , and TXB 2, in cultured cells using liquid chromatography/tandem mass spectrometry (LC/MS/MS), and we validated the LC/MS/MS methods. A solid-phase extraction cartridge was used for extraction of prostanoids. The prostanoids were separated by a C 18 column with an isocratic flow of acetonitrile/water/acetic acid (40:60:0.1, v/v/v). Calibration curves of extracellular measurement for the prostanoids were linear in the range from 0.1 to 100ng/mL (r 2>0.999), and those of intracellular measurement were linear in the range from 0.05 to 50ng (r 2>0.999). Validation assessment showed that both methods of extracellular and intracellular measurements were highly reliable with good accuracy and precision. We also applied the methods to human airway epithelial Calu-3 cells and human lung adenocarcinoma epithelial A549 cells.

Original languageEnglish
Pages (from-to)3378-3385
Number of pages8
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume879
Issue number30
DOIs
Publication statusPublished - 2011 Nov 15

Keywords

  • Intracellular measurement
  • LC/MS/MS
  • Prostanoid

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

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