Zeaxanthin (Zea) and lutein are the only dietary carotenoids that accumulate in the macular region of the retina and lens. It was proposed that these carotenoids protect these tissues against photooxidative damage. Few plant foods are enriched in Zea, and information about the bioavailability of Zea from these foods and its accumulation in ocular tissues is limited. The amounts of free Zea and its mono- and diesters were measured for several plant foods that have relatively high concentrations of this xanthophyll. Wolfberry had the greatest concentration of Zea with a diester that accounts for 95% of the total. Free, mono-, and diesters of Zea were present in orange and red peppers, whereas only Zea monoesters were detected in squash. Zea esters were partially hydrolyzed by carboxyl ester lipase (CEL) during simulated digestion. The efficiency of micellarization was dependent on speciation with combined means of free Zea, Zea monoesters, Zea diesters from the digested foods of 81 +/- 8, 44 +/- 5, and 11 +/- 4%, respectively. When exposed to micelles generated during digestion of the test foods, Zea uptake by Caco-2 cells was proportional to the medium content (11-14%). Free Zea was the most abundant form in Caco-2 cells, although Zea monoesters also were detected (<8 +/- 0.7% vs. free Zea). CEL enhanced Zea uptake from micelles (12.3-fold; P < 0.05) by hydrolyzing Zea esters. After cell uptake, concentrations of free and monoesterified Zea remained relatively stable. These data suggest that dietary Zea esters are hydrolyzed by CEL during the small intestinal phase of digestion and that this conversion enhances Zea bioavailability.