Preparation and assay of monohydroxy-eicosatetraenoic acids

J. M. Boeynaems, A. R. Brash, J. A. Oates, W. C. Hubbard

Research output: Contribution to journalArticle

Abstract

5-, 8-, 9-, 11-, 12-, and 15-hydroxy-eicosatetraenoic acids (HETEs) were prepared from arachidonic acid by reaction with H2O2 in the presence of Cu2+ ions. They were separated by high-performance liquid chromatography on silica gel (μPorasil), using a linear solvent gradient from hexane to chloroform: only the 8- and 9-isomers were not resolved. Multi-milligram quantities of highly purified HETEs could be easily generated by this method, which thus provides a useful tool to study the biological activity of these compounds. Octadeuterated analogs of HETEs prepared from octadeuterated arachidonic acid by this procedure were suitable for use as internal standards in stable isotope dilution assays, by combined gas chromatography and mass spectrometry, with selected ion monitoring. The detection limit of the HETEs was less than 1 ng.

Original languageEnglish (US)
Pages (from-to)259-267
Number of pages9
JournalAnalytical biochemistry
Volume104
Issue number2
DOIs
StatePublished - May 15 1980

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ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Boeynaems, J. M., Brash, A. R., Oates, J. A., & Hubbard, W. C. (1980). Preparation and assay of monohydroxy-eicosatetraenoic acids. Analytical biochemistry, 104(2), 259-267. https://doi.org/10.1016/0003-2697(80)90073-1