Rapid determination of soluble epoxide hydrolase inhibitors in rat hepatic microsomes by high-performance liquid chromatography with electrospray tandem mass spectrometry

Takaho Watanabe, Bruce D. Hammock

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

A rapid and reliable electrospray tandem mass spectrometric method for soluble epoxide hydrolase (sEH) inhibitors in rat hepatic microsomes is described. Four synthesized sEH inhibitors were extracted from rat hepatic microsomes with ethyl acetate and were determined by HPLC using positive ion electrospray tandem mass spectrometry within 7 min. The relationship between signal intensity and concentration of sEH inhibitors was linear over the concentration range of 2.0 to 500 ng/mL per 5-μL injection with the use of a noncoeluting internal standard with a similar chemical structure. The intraassay precision was less than 12.4% relative standard deviation and accuracy ranged from -7.0 to 11.3% deviation from the theoretical values with five duplicate assays. The recovery of sEH inhibitors from rat hepatic microsomes, fortified at levels of 50, 100, and 250 ng/mL, averaged 74.2-107.7% with a RSD of 2.1-7.6%. This method was successfully applied to the quantification of residual sEH inhibitors in rat hepatic microsomes without interference.

Original languageEnglish (US)
Pages (from-to)227-234
Number of pages8
JournalAnalytical Biochemistry
Volume299
Issue number2
DOIs
StatePublished - Dec 15 2001

Keywords

  • Electrospray ionization
  • Inhibitor
  • Liquid chromatography
  • Quantification
  • Rat hepatic microsomes
  • Soluble epoxide hydrolase
  • Tandem mass spectrometry

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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