Gradient reversed-phase high-performance liquid chromatographic separation of naturally occurring retinoids.
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.
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004224:Diterpenes; D005260:Female; D006801:Humans; D007668:Kidney / Q000032:analysis; D008099:Liver / Q000032:analysis*; D008114:Liver Neoplasms, Experimental / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D012176:Retinoids / Q000032:analysis*; D000084562:Retinyl Esters; D013552:Swine; D014020:Tissue Extracts / Q000032:analysis; D014801:Vitamin A / Q000031:analogs & derivatives / Q000032:analysis
D004224:Diterpenes; D012176:Retinoids; D000084562:Retinyl Esters; D014020:Tissue Extracts; D014801:Vitamin A; C014794:retinol palmitate