Stereoselective determination of vigabatrin enantiomers in human plasma by high performance liquid chromatography using UV detection

Valentina Franco, Iolanda Mazzucchelli, Cinzia Fattore, Roberto Marchiselli, Giuliana Gatti, Emilio Perucca

Research output: Contribution to journalArticle

Abstract

A rapid and simple high-performance liquid chromatographic method for the determination of the R-(-)- and S-(+)-enantiomers of the antiepileptic drug vigabatrin in human plasma is described. After adding the internal standard (1-aminomethyl-cycloheptyl-acetic acid), plasma samples (200 μL) are deproteinized with acetonitrile and the supernatant is derivatized with 2,4,6 trinitrobenzene sulfonic acid (TNBSA). Separation is achieved on a reversed-phase cellulose-based chiral column (Chiralcel-ODR, 250 mm × 4.6 mm i.d.) using 0.05 M potassium hexafluorophosphate (pH 4.5)/acetonitrile/ethanol (50:40:10 vol/vol/vol) as mobile phase at a flow-rate of 0.9 mL/min. Chromatographic selectivity is improved by concentrating the derivatives on High Performance Extraction Disk Cartridges prior to injection. Detection is at 340 nm. Calibration curves are linear (r2 ≥ 0.999) over the range of 0.5-40 μg/mL for each enantiomer, with a limit of quantification of 0.5 μg/mL for both analytes. The assay is suitable for therapeutic drug monitoring and for single-dose pharmacokinetic studies in man.

Original languageEnglish
Pages (from-to)63-67
Number of pages5
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume854
Issue number1-2
DOIs
Publication statusPublished - Jul 1 2007

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Keywords

  • Enantiomers
  • Enantioselective assay
  • HPLC
  • UV detection
  • Vigabatrin

ASJC Scopus subject areas

  • Biochemistry

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