Simultaneous quantification of retinol, retinal, and retinoic acid isomers by high-performance liquid chromatography with a simple gradiation

Michiko Miyagi, Hirokazu Yokoyama, Haruko Shiraishi, Michinaga Matsumoto, Hiromasa Ishii

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28 Citations (Scopus)


A new method of high-performance liquid chromatography (HPLC) analysis to quantify isomers of retinol, retinal and retinoic acid simultaneously was established. The HPLC system consisted of a silica gel absorption column and a linear gradient with two kinds of solvents containing n-Hexane, 2-propanol, and glacial acetic acid in different ratios. It separated six retinoic acid isomers (13-cis, 9-cis, all-trans, all-trans-4-oxo, 9-cis-4-oxo, 13-cis-4-oxo), three retinal isomers (13-cis-, 9-cis-, and all-trans) and two retinol isomers (13-cis- and all-trans). Human serum samples were subjected to this HPLC analysis and at least, all-trans retinol, 13-cis retinol, and all-trans retinoic acid were detectable. This HPLC system is useful for evaluating retinoic acid formation from retinol via a two-step oxidation pathway. Moreover, it could be applied to monitoring the concentrations of various retinoids, including all-trans retinoic acid in human sera.

Original languageEnglish
Pages (from-to)365-368
Number of pages4
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Issue number2
Publication statusPublished - 2001 Jun 15



  • Retinal
  • Retinoic acid
  • Retinol
  • Vitamins

ASJC Scopus subject areas

  • Chemistry(all)

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