Simultaneous determination of retinol, α-tocopherol and retinyl palmitate in plasma of premature newborns by reversed-phase high-performance liquid chromatography

Angela Bortolotti, Giorgio Lucchini, Maria Monica Barzago, Fabio Stellari, Maurizio Bonati

Research output: Contribution to journalArticle

Abstract

A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of retinol, α-tocopherol and retinyl palmitate in plasma. Plasma containing an internal standard (tocol) was deproteinized with ethanol, then extracted with n-hexane. The organic layer was removed and evaporated under a nitrogen stream, and chromatographed on a reversed-phase RP-18 column using a water/acetonitrile-ethyl acetate/2-propanol gradient solvent system over 15 min at 305 nm. The recovery exceeded 93%. The detection limit was 0.1 μg/ml for retinol, 1.3 μg/ml for α-tocopherol and 0.95 μg/ml for retinyl palmitate. The reproducibility, precision (expressed as coefficients of variation) and accuracy were less than 8% for all analytes. The small sample requirement, the simplicity of extraction, the short run-time and the good reproducibility make this procedure particularly useful for monitoring retinol and α-tocopherol supplementation in premature newborns.

Original languageEnglish
Pages (from-to)313-317
Number of pages5
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume617
Issue number2
DOIs
Publication statusPublished - Aug 11 1993

ASJC Scopus subject areas

  • Chemistry(all)
  • Clinical Biochemistry
  • Molecular Medicine

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