Simultaneous liquid chromatographic determination of carotenoids, retinoids, and tocopherols in human buccal mucosal cells.

Epidemiological and experimental data have suggested that some micronutrients, including various carotenoids, retinoids, and alpha-tocopherol, may have chemopreventive activity against certain types of human cancer. In order to define the role of these micronutrients in cancer prevention, it is necessary to measure their concentrations in the target tissues, since these are critical to the chemopreventive effect. We have developed a sensitive and reproducible high-performance liquid chromatography procedure for the simultaneous determination of lutein, zeaxanthin, beta-cryptoxanthin, lycopene, alpha-carotene, beta-carotene, cis-beta-carotene, retinol, retinyl palmitate, alpha-tocopherol, and gamma-tocopherol in an easily accessible human target tissue, the buccal mucosal cells. This procedure used a gradient of two mobile phases which consisted of acetonitrile, tetrahydrofuran, methanol, 1% ammonium acetate, and butylated hydroxytoluene (in v/v/v/v/w): mobile phase A, 85:5:5:5:0.05; mobile phase B, 55:35:5:5:0.05. The run time, including reequilibration, was 47 min: from 100% A to 100% B in 27 min, remaining at that mobile phase for 10 min, then back to 100% A at 43 min at a constant flow rate of 1.3 ml/min. The high-performance liquid chromatography effluent was monitored at 300, 325, and 452 nm for tocopherols, retinoids, and carotenoids, respectively, with a photodiode array detector. The average recovery was 83% for lycopene and > 92% for others. The precision of the assay for all the compounds was less than 10% in a 1-month period. The micronutrients were extracted by incubating the cells with protease, followed by vortex mixing with 1% sodium dodecyl sulfate in ethanol containing 0.1% butylated hydroxytoluene, and, finally, hexane extraction.(ABSTRACT TRUNCATED AT 250 WORDS)