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

研究成果: Article査読

11 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)3378-3385
ページ数8
ジャーナルJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
879
30
DOI
出版ステータスPublished - 2011 11 15

ASJC Scopus subject areas

  • 分析化学
  • 生化学
  • 臨床生化学
  • 細胞生物学

フィンガープリント

「Quantification of intracellular and extracellular prostanoids stimulated by A23187 by liquid chromatography/electrospray ionization tandem mass spectrometry」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル