Rapid and simultaneous determination of efavirenz, 8-hydroxyefavirenz, and 8,14-dihydroxyefavirenz using LC-MS-MS in human plasma and application to pharmacokinetics in healthy volunteers

Kwon Bok Kim, Hyunmi Kim, Fen Jiang, Chang Woo Yeo, Soo Kyung Bae, Zeruesenay Desta, Jae Gook Shin, Kwang Hyeon Liu

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10 Scopus citations

Abstract

We developed a rapid and sensitive method for determining efavirenz, 8-hydroxyefavirenz, and 8,14-dihydroxyefavirenz in human plasma simultaneously using liquid chromatography-tandem mass spectrometry (LC-MS-MS). Three compounds and ritonavir, an internal standard, were extracted from plasma using ethyl acetate in the presence of 0.1 M sodium carbonate after incubation of β-glucuronidase (500 U). After drying the organic layer, the residue was reconstituted in mobile phase (acetonitrile:20 mM ammonium acetate, 90:10, v/v) and injected onto a reversed-phase C18 column. The isocratic mobile phase was eluted at 0.2 mL min-1. The ion transitions monitored in multiple reaction-monitoring mode were m/z 314 → 244, 330 → 258, 346 → 262, and 721 → 296 for efavirenz, 8-hydroxyefavirenz, 8,14-dihydroxyefavirenz, and ritonavir, respectively. The retention time is 1.93, 1.70, 1.52, and 1.82 min for efavirenz, 8-hydroxyefavirenz, 8,14-dihydroxyefavirenz, and ritonavir, respectively. The coefficients of variation of the assay precision were less than 10.7%, and the accuracy was 90-111%. The lower limits of quantification (LLOQ) were 5 ng mL-1 for efavirenz and 8-hydroxyefavirenz. This method was used to measure the plasma concentrations of efavirenz and its metabolites from healthy volunteers after a single 600 mg oral dose of efavirenz. This analytical method is a very rapid, sensitive, and accurate to determine the pharmacokinetic profiles of efavirenz including its metabolites.

Original languageEnglish
Pages (from-to)263-271
Number of pages9
JournalChromatographia
Volume73
Issue number3-4
DOIs
StatePublished - Feb 2011

Keywords

  • 8,14-Dihydroxyefavirenz
  • 8-Hydroxyefavirenz
  • Column liquid chromatography-tandem mass spectrometry
  • Efavirenz
  • Human plasma

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