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  • Title: Gradient reversed-phase high-performance liquid chromatographic separation of naturally occurring retinoids.
    Author: Furr HC, Amédée-Manesme O, Olson JA.
    Journal: J Chromatogr; 1984 Aug 10; 309(2):299-307. PubMed ID: 6480781.
    Abstract:
    A gradient reversed-phase high-performance liquid chromatographic technique is described for the facile separation and quantitation of the naturally occurring retinoids: retinoic acid, retinol, and retinyl esters. An octadecylsilane column (Waters mu Bondapak C18) is used, with gradient elution from methanol--water (80:20) (solvent A) to 70% or 100% methanol--tetrahydrofuran (50:50) (solvent B) at 2.0 ml/min; detection is by absorbance at 325 nm. Analysis can be completed, with return to starting conditions, in 25-30 min. The method is inherently flexible: retinyl esters can be eluted as a group, with little resolution, by gradient to 100% solvent B, or mostly resolved by gradient to 70% solvent B; separation of retinoids more polar than retinoic acid can be achieved by use of greater proportions of water in solvent A. The separation of vitamin A compounds from extracts of human, rat, and pig liver and from rat kidney by this technique is described.
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