Simultaneous quantification of prostaglandins, isoprostane and thromboxane in cell-cultured medium using gas chromatography-mass spectrometry

Hiroki Tsukamoto, Takanori Hishinuma, Tsuyoshi Mikkaichi, Hironori Nakamura, Tohru Yamazaki, Yoshihisa Tomioka, Michinao Mizugaki

Research output: Contribution to journalArticlepeer-review

38 Citations (Scopus)

Abstract

We have developed a simultaneous quantification method for prostaglandin (PG) E2, PGD2, PGF, 8-epi-PGF, 6-keto-PGF and thromboxane (TX) B2. Using [3,3,4,4-2H4]PGE2, [3,3,4,4-2H4]PGD2, [3,3,4,4-2H4]8-epi-PGF, [3,3,4,4-2H4]PGF, [3,3,4,4-2H4]6-keto-PGF and [18,18,19,19-2H4]TXB2 as internal standards (I.S.), the eicosanoids and their I.S. were simultaneously extracted by solid-phase extraction from cell-cultured medium, derivatized to methyl ester/methoxim/tert.-butyldimethylsilyl ether derivatives and analyzed using gas chromatography-mass spectrometry in the selected ion monitoring mode. The accuracy for the added eicosanoids ranged from 92 to 113%, and coefficients of variation ranged from 0.1 to 12.2%. Increased eicosanoids in RAW264.7 and U937 cells stimulated by lipopolysaccharide were suppressed by NS-398 and indometacin. This simultaneous quantification method can be applied routinely for assaying eicosanoids in vitro.

Original languageEnglish
Pages (from-to)205-214
Number of pages10
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume774
Issue number2
DOIs
Publication statusPublished - 2002 Jul 15

Keywords

  • Isoprostane
  • Prostaglandins
  • Thromboxane

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

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