An improved HPLC Method for the Separation of Fourteen Carotenoids, Including 15-/13- and 9-CIS-β-Carotene Isomers, Phytoene and Phytofluene

The isocratic separation of 14 carotenoids, as well as retinol, retinyl acetate, retinyl palmitate, α-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, β-apo-8′ carotenal, β-cryptoxanthin, echinenone, lycopene, γ-carotene, α-carotene, β-carotene, 9-cis-β-carotene, 15-cis-/13-cis-β-carotene, phytoene and phytofluene. The separation of lutein and zeaxanthin is obtained simply by changing the mobile phase to acetonitrile:methanol (85:15).