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An improved hplc method for the separation of fourteen carotenoids including 15 cis beta carotene 13 cis beta carotene and 9 cis beta carotene isomers phytoene and phytofluene



An improved hplc method for the separation of fourteen carotenoids including 15 cis beta carotene 13 cis beta carotene and 9 cis beta carotene isomers phytoene and phytofluene



Journal of Liquid Chromatography 14(13): 2457-2476



The isocratic separation of 14 carotenoids, as well as retinol, retinyl acetate, retinyl palmitate, .alpha.-tocopherol and tocopherol acetate, is accomplished in 12 minutes, using a Spheri-5-ODS column and acetonitrile:dichloromethane:methanol (70:20:10) as mobile phase, with two-channel, programmable multiwavelength detection. The carotenoids separated are as follows: lutein/zeaxanthin, canthaxanthin, .beta.-apo-8'carotenal, .beta.-cryptoxanthin, echinenone, lycopene, .gamma.-carotene, .alpha.-carotene, .beta.-carotene, 9-cis-.beta.-carotene, 15-cis-/13-cis-.beta.-carotene, phytoene and phytofluene. The separation of lutein and zeaxanthin is obtained simply by changing the mobile phase to acetonitrile:methanol (85:15). The separation of the cis-isomers of .beta.-carotene is maintained, in spite of changes in the proportions and even after 250 hours of analysis. This chromatographic separation is not dependent on temperature (within a range of 15-37.degree.C), on the injection solvent (THF, EtOH or the mobile phase) or flow rate (in a range of 1.3-2.5 ml/min). The identification of these compounds has been assayed in both vegetable samples and serum.

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