Improved enantioselective assay for the determination of fluoxetine and norfluoxetine enantiomers in human plasma by liquid chromatography

Giuliana Gatti, Ilaria Bonomi, Roberto Marchiselli, Cinzia Fattore, Edoardo Spina, Gabriella Scordo, Roberta Pacifici, Emilio Perucca

Research output: Contribution to journalArticle

Abstract

A simple and innovative assay is described which allows the chiral separation of the four enantiomers of fluoxetine and norfluoxetine, with performance characteristics adequate for therapeutic drug monitoring. The assay requires liquid-liquid extraction into acetonitrile/n-hexane/isopropylic alcohol and re-extraction into phosphoric acid for clean-up. The acidic layer is injected onto the HPLC system after filtering. Separation of the analytes is achieved with a Chiralcel ODR column and a mobile phase consisting of potassium hexafluorophosphate/acetonitrile. Detection is made by ultraviolet absorbance at 227 nm. Standard curves are linear for each enantiomer (r2≥0.992) over the range of 10-1000 ng/ml with a limit of quantification of 10 ng/ml for each enantiomer. Within-day and between-day CV% are ≤10% for each enantiomer.

Original languageEnglish
Pages (from-to)375-383
Number of pages9
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume784
Issue number2
DOIs
Publication statusPublished - Feb 5 2003

Keywords

  • Enantiomer separation
  • Fluoxetine
  • Norfluoxetine

ASJC Scopus subject areas

  • Biochemistry

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