SEPARATION AND IDENTIFICATION OF CAROTENOIDS AND THEIR OXIDATION-PRODUCTS IN THE EXTRACTS OF HUMAN PLASMA

Eighteen carotenoids as well as vitamin A and two forms of vitamin E (gamma- and alpha-tocopherol) have been separated from extracts of human plasma by high-performance liquid chromatography (HPLC) on reversed-phase and silica-based nitrile-bonded columns. In the order of chromatographic elution on a C18 reversed-phase column, the carotenoids were identified as (3R,3'R,6'R)-beta,epsilon-carotene-3,3'-diol [(3R,3'R,6'-lutein], (3R,3'R)-beta,beta-carotene-3,3'-diol [(3R,3'R)-zeaxanthin], 5,6-dihydroxy-5,6-dihydro-psi,psi-carotene, 3-hydroxy-2",3'-didehydro-beta,epsilon-carotene,beta,epsilon-caroten-3-ol, 3-hydroxy-beta-carotene, psi,psi-carotene, 7,8-dihydro-psi,psi-carotene, beta,psi-carotene, 7,8,7',8'-tetrahydro-psi,psi-carotene, beta,epsilon-carotene, beta,beta-carotene, 7,8,11,12,7',8'-hexahydro-psi,psi-carotene and 7,8,11,12,7',8',11',12'octahydro-psi,psi-carotene. The polar carotenoids which eluted in the vicinity of lutein and were unresolved on the C18 column, have been separated on a nitrile-bonded column employing isocratic HPLC conditions. In the order of elution, the carotenoids were epsilon,epsilon-carotene-3,3'-dione, 3'-hydroxy-epsilon,epsilon-caroten-3-one, 5,6-dihydroxy-5,6-dihydro-psi,psi-carotene, 3-hydroxy-beta,epsilon-caroten-3'-one, (all-E,3R,3'R,6'R)-lutein, (all-E,3R,3'R)-zeaxanthin, and (all-E,3R,3'S,6'R)-beta,epsilon-carotene-3,3'-diol (3'-epilutein) followed by several geometrical isomers of lutein and zeaxanthin.