As in humans and monkeys, lutein [(3R,3′R,6′R)-βε, -carotene-3,3′-diol] and zeaxanthin [a mixture of (3R,3′R)-β,β-carotene-3,3′diol and (3R,3′S-meso)-β,β-carotene-3,3′-diol] are found in substantial amounts in the retina of the Japanese quail Coturnix japonica. This makes the quail retina an excellent non-primate small animal model for studying the metabolic transformations of these important macular carotenoids that are thought to play an integral role in protection against light-induced oxidative damage such as that found in age-related macular degeneration (AMD). In this study, we first identified the array of carotenoids present in the quail retina using C30-HPLC coupled with inline mass-spectral and photodiode-array detectors. In addition to dietary lutein (2.1%) and zeaxanthin (11.8%), we identified adonirubin (5.4%), 3′-oxolutein (3.8%), meso-zeaxanthin (3.0%), astaxanthin (28.2%), galloxanthin (12.2%), ε,ε-carotene (18.5%), and β-apo-2′-carotenol (9.5%) as major ocular carotenoids. We next used deuterium-labeled lutein and zeaxanthin as dietary supplements to study the pharmacokinetics and metabolic transformations of these two ocular pigments in serum and ocular tissues. We then detected and quantitated labeled carotenoids in ocular tissue using both HPLC-coupled mass spectrometry and non-invasive resonance Raman spectroscopy. Results indicated that dietary zeaxanthin is the precursor of 3′-oxolutein, β-apo-2′-carotenol, adonirubin, astaxanthin, galloxanthin, and ε, ε-carotene, whereas dietary lutein is the precursor for mesozeaxanthin. Studies also revealed that the pharmacokinetic patterns of uptake, carotenoid absorption, and transport from serum into ocular tissues were similar to results observed in most human clinical studies.
KeywordsCarotenoid; Retina; Japanese quail; Lutein; Zeaxanthin; HPLC; APCI-MS Epidemiological and clinical studies in humans have generally demonstrated that there is an inverse relationship between nutritional consumption of antioxidants and prevalence of old age ocular disorders such as age-related macular degeneration (AMD) and cataract (1,2). The carotenoids lutein and zeaxanthin are notable examples because they are specifically concentrated in ocular tissues at high concentrations, and they are excellent absorbers of phototoxic blue light and reactive oxygen species (3,4). Studies using data from the Eye Disease Case Control (EDCC) Study Group have demonstrated that individuals with high blood levels of lutein and zeaxanthin and high consumption of foods rich in these same carotenoids had significantly lower risk of exudative AMD, and several follow up studies including the Age- † This work was supported by National Institute of Health Grant EY-11600 and by Research to Prevent Blindness, Inc. (New York, NY). In the human retina, dietary lutein [(3R,3′R,6′R)-β,ε-carotene-3,3′-diol] and zeaxanthin [(3R, 3′R)-β,β-carotene-3,3′-diol] are unevenly distributed in the retina, with millimolar concentrations found at the fovea with a lutein:zeaxanthin ratio of 1:2 and a...