Simultaneous analysis of individual carotenoids, retinol, retinyl esters, and tocopherols in serum by isocratic non-aqueous reversed-phase HPLC
1993
Barua, A.B. | Furr, H.C. | Janick-Buckner, D. | Olson, J.A.
A sensitive high-performance reversed-phase liquid chromatographic assay which utilizes isocratic elution and detection by two detectors has been developed to measure simultaneously several carotenoids, retinol, retinyl esters, and alpha- and gamma-tocopherols in serum. Although 100 microliter of serum or plasma is used routinely, as little as 10 microliter can be analyzed. Typical lower limits of detection (5:1 signal/noise ratio) are 0.2 pmol for retinol, 6 pmol for alpha-tocopherol, 1 pmol for beta-carotene, and 2 pmol for retinyl palmitate. Although lutein and zeaxanthin were only partially resolved from each other, all other major serum carotenoids including alpha-carotene from beta-carotene and alpha-cryptoxanthin from beta-cryptoxanthin were completely separated. Retinyl hexanoate and beta-apo-8'-carotenyl decanoate are convenient, readily prepared internal standards for this assay. Relative retention indices, based on elution times of alkylphenone standards, proved useful both for characterization of compounds and for comparison between columns.
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