Lutein, zeaxanthin, and the macular pigment.

The predominant carotenoids of the macular pigment are lutein, zeaxanthin, and meso-zeaxanthin. The regular distribution pattern of these carotenoids within the human macula indicates that their deposition is actively controlled in this tissue. The chemical, structural, and optical characteristics of these carotenoids are described. Evidence for the presence of minor carotenoids in the retina is cited. Studies of the dietary intake and serum levels of the xanthophylls are discussed. Increased macular carotenoid levels result from supplementation of humans with lutein and zeaxanthin. A functional role for the macular pigment in protection against light-induced retinal damage and age-related macular degeneration is discussed. Prospects for future research in the study of macular pigment require new initiatives that will probe more accurately into the localization of these carotenoids in the retina, identify possible transport proteins and mechanisms, and prove the veracity of the photoprotection hypothesis for the macular pigments.

Macular pigment in donor eyes with and without AMD: a case-control study.

PURPOSE: To determine whether there is an association between the density of macular pigment in the human retina and the risk of age-related macular degeneration (AMD). METHODS: Retinas from 56 donors with AMD and 56 controls were cut into three concentric regions centered on the fovea. The inner, medial, and outer regions covered the visual angles 0 degrees to 5 degrees, 5 degrees to 19 degrees, and 19 degrees to 38 degrees, respectively. The amounts of lutein (L) and zeaxanthin (Z) extracted from each tissue sample were determined by high-performance liquid chromatography. RESULTS: L and Z levels in all three concentric regions were less, on average, for the AMD donors than for the controls. The differences decreased in magnitude from the inner to medial to outer regions. The lower levels found in the inner and medial regions for AMD donors may be attributable, in part, to the disease. Comparisons between AMD donors and controls using the outer (peripheral) region were considered more reliable. For this region, logistic regression analysis indicated that those in the highest quartile of L and Z level had an 82% lower risk for AMD compared with those in the lowest quartile (age- and sex-adjusted odds ratio = 0.18, 95% confidence interval = 0.05-0.64). CONCLUSIONS: The results are consistent with a theoretical model that proposes an inverse association between risk of AMD and the amounts of L and Z in the retina. The results are inconsistent with a model that attributes a loss of L and Z in the retina to the destructive effects of AMD.

Lutein and zeaxanthin in the eyes, serum and diet of human subjects.

Inverse associations have been reported between the incidence of advanced, neovascular, age-related macular degeneration (AMD) and the combined lutein (L) and zeaxanthin (Z) intake in the diet, and L and Z concentration in the blood serum. We suggest that persons with high levels of L and Z in either the diet or serum would probably have, in addition, relatively high densities of these carotenoids in the macula, the so-called 'macular pigment'. Several lines of evidence point to a potential protective effect by the macular pigment against AMD. In this study we examined the relationship between dietary intake of L and Z using a food frequency questionnaire; concentration of L and Z in the serum, determined by high-performance liquid chromatography, and macular pigment optical density, obtained by flicker photometry. Nineteen subjects participated. We also analysed the serum and retinas, as autopsy samples, from 23 tissue donors in order to obtain the concentration of L and Z in these tissues. The results reveal positive, though weak, associations between dietary intake of L and Z and serum concentration of L and Z, and between serum concentration of L and Z and macular pigment density. We estimate that approximately half of the variability in the subjects' serum concentration of L and Z can be explained by their dietary intake of L and Z, and about one third of the variability in their macular pigment density can be attributed to their serum concentration of L and Z. These results, together with the reported associations between risk of AMD and dietary and serum L and Z, support the hypothesis that low concentrations of macular pigment may be associated with an increased risk of AMD.

A one year study of the macular pigment: the effect of 140 days of a lutein supplement.

A low density of macular pigment may represent a risk factor for age-related macular degeneration (AMD) by permitting greater blue light damage. This study was carried out to determine the effects on macular pigment optical density of dietary supplementation with lutein, one of the pigment constituents. Two subjects consumed lutein esters, equivalent to 30 mg of free lutein per day, for a period of 140 days. Macular pigment optical density was determined by heterochromatic flicker photometry before, during, and after the supplementation period. Serum lutein concentration was also obtained through the analysis of blood samples by high-performance liquid chromatography. Twenty to 40 days after the subjects commenced taking the lutein supplement, their macular pigment optical density began to increase uniformly at an average rate of 1.13+/-0.12 milliabsorbance units/day. During this same period, the serum concentration of lutein increased roughly tenfold, approaching a steady state plateau. The optical density curve eventually levelled off 40 to 50 days after the subjects discontinued the supplement. During the same 40 to 50 days, the serum concentration returned to baseline. Thereafter, little or no decrease in optical density was observed. The mean increases in the macular pigment optical density were 39% and 21% in the eyes of the two subjects respectively. In conclusion, the modest period of supplementation has been estimated to have produced in the subjects a 30 to 40% reduction in blue light reaching the photoreceptors, Bruch's membrane, and the retinal pigment epithelium, the vulnerable tissues affected by AMD.

Distribution of lutein and zeaxanthin stereoisomers in the human retina.

The distribution of macular pigment stereoisomers in the human retina has been mapped and a pathway to account for the presence of the non-dietary carotenoid, meso-zeaxanthin, is proposed. Adult neural retinas were cut into three concentric areas centered on the fovea, and the extracted carotenoids were analysed and purified by high-performance liquid chromatography. The dicarbamate or dibenzoate derivatives of the collected zeaxanthin fractions for each tissue sample were further analysed by HPLC to determine their stereoisomer composition. Whole retinas from infant eyes were similarly analysed. The results show that, relative to zeaxanthin, the concentration of lutein in the adult neural retina increases with radial distance from the fovea while that of meso-zeaxanthin decreases. Infant retinas were found to have more lutein and less meso-zeaxanthin, relative to zeaxanthin, than adult retinas. Small quantities of (3S, 3'S)-zeaxanthin were also found in the adult retina, particularly in the macula. It is proposed that lutein and zeaxanthin are transported into an individual's retina in the same proportions found in his or her blood serum. Some of the lutein is then converted into meso-zeaxanthin, primarily in the macula, by a mechanism which is less developed in infants than adults.

Stereochemistry of the human macular carotenoids.

PURPOSE: To complete identification of the major components of the human macular pigment. METHODS: Chemical ionization mass spectra of the macular pigment components were obtained and compared with those of zeaxanthin and lutein standards. A comparison was also made using chiral column high-performance liquid chromatography, which is capable of resolving individual stereoisomers of these carotenoids. Zeaxanthin and lutein from human blood plasma were similarly analyzed. RESULTS: The mass spectrometry data supported earlier work in which high-performance liquid chromatography, UV-visible spectrometry and chemical modification showed that the macular pigment comprises two carotenoids with identical properties to those of zeaxanthin and lutein. Chiral column chromatography showed that the "zeaxanthin" fraction is a mixture of two stereoisomers, zeaxanthin itself [(3R,3'R)-beta,beta-Carotene-3,3'-diol] and meso-zeaxanthin [(3R,3'S)-beta,beta-Carotene-3,3'-diol]. The other fraction is the single stereoisomer, lutein [(3R,3'R,6'R)-beta,epsilon-Carotene-3,3'-diol]. In human blood plasma, only zeaxanthin and lutein were found. CONCLUSIONS: The results strongly suggest that meso-zeaxanthin results from chemical processes within the retina. Noting that lutein exceeds zeaxanthin in plasma but that the combined zeaxanthin stereoisomers exceed lutein in the retina, the possibility was considered that meso-zeaxanthin is a conversion product derived from retinal lutein. Under nonphysiologic conditions, the authors demonstrate that a base-catalyzed conversion of lutein to zeaxanthin yields only the meso-(3R,3'S) stereoisomer.

Optical density spectra of the macular pigment in vivo and in vitro.

A precise relative optical density spectrum of the macular pigment, based upon its dichroic properties, was determined. The spectrum proved essentially identical to that of liposome-bound zeaxanthin and lutein, a system duplicating the macular pigment and its environment. Substantial agreement was also found with the spectra of Wyszecki and Stiles (1982, Color science: Concepts and methods, quantitative data and formulae. New York: Wiley) and Vos (1972, Institute for Perception, RVO-TNO, IZF 1972-17, Soesterberg, The Netherlands), and the latter is recommended as a standard. For 7 subjects, the pigment density spectrum derived from foveal and extra-foveal sensitivities was compared with the dichroism-based spectrum. Results indicated that the pigment is described by a common distribution of molecular orientations for all subjects.

Photoreceptor-specific efficiencies of beta-carotene, zeaxanthin and lutein for photopigment formation deduced from receptor mutant Drosophila melanogaster.

Drosophila rearing media had only beta-carotene, zeaxanthin or lutein as precursors for photopigment chromophores. Zeaxanthin and lutein are potentially optimum sources of the 3-hydroxylated retinoids of visual and accessory photopigments. Mutants made the electroretinogram in white (w) eyes selective for compound eye photoreceptors R1-6, R7 and R8: R1-6 dominates w's electroretinogram; R7/8 generates w;ora's (ora = outer rhabdomeres absent); R8 generates w sev;- ora's (sev = sevenless). Microspectrophotometry revealed R1-6's visual pigment. In w, all 3 carotenoids yielded monotonic dose-responses for sensitivity or visual pigment. An ultraviolet sensitivity peak from R1-6's sensitizing pigment was present at high but not low doses. In w;ora, all 3 carotenoids gave similar spectra dominated by R7's high ultraviolet sensitivity. For w sev;ora, all spectra were the shape expected for R8, peaking around 510 nm. The sensitivity dose-response was at its ceiling except for low doses in w;ora and zero supplementation in w sev;ora. Hence, without R1-6, most of our dose range mediated maximal visual pigment formation. In Drosophila, beta-carotene, zeaxanthin and lutein mediate the formation of all major photopigments in R1-6, R7 and R8.

Analysis of the macular pigment by HPLC: retinal distribution and age study.

High performance liquid chromatography (HPCL) has been employed to study the distribution throughout the human retina of zeaxanthin and lutein, the two major components of the macular pigment. Differences between individuals have also been studied with a view to uncovering possible age-related effects. Both pigments were detected in prenatal eyes (approximately 20 weeks gestation) but did not form a visible yellow spot. Generally they were not easily discernible until about 6 months after birth. For 87 donors between the ages of 3 and 95, no dependence on age was observed in the quantity of either pigment. For approximately 90% of these, zeaxanthin was dominant. For the remaining 10%, as well as for the seven youngest donors, all below the age of 2, and in prenatal eyes, lutein was the major pigment. In individual retinas, the lutein:zeaxanthin ratio increased from an average of approximately 1:2.4 in the central 0-0.25 mm to over 2:1 in the periphery (8.7-12.2 mm). The variation in this ratio with eccentricity was linearly correlated with the corresponding rod:cone ratio. A selective mechanism of uptake, which results in cones and rods preferentially acquiring zeaxanthin and lutein, respectively, could explain this correlation.

Preliminary identification of the human macular pigment.

The human macular pigment has been found to be composed to two chromatographically separable components, which are tentatively identified as lutein [(3R,3'R,6'R)-beta,epsilon-Carotene-3,3'-diol] and zeaxanthin [(3R,3'R)-beta,beta-Carotene-3,3'-diol]. Chromatograms of retinal extracts, obtained by HPLC on three different stationary phases, were found to match those obtained with mixtures of lutein and zeaxanthin standards. Identical retention times were confirmed by coinjection of each of the isolated components with the appropriate standard. U.V.-visible spectra of the purified components were identical in all respects with those of lutein and zeaxanthin. Further support for our identification was obtained by the preparation and chromatographic comparison of derivatives of the macular pigment and of the standards.

Macular pigment in Henle fiber membranes: a model for Haidinger's brushes.

The dichroic properties of lutein, presumed to be the macular pigment, are demonstrated and the structure of Henle fiber membranes is discussed. A consequence of the pigment molecules being incorporated into the bilipid components of these membranes is shown to be the production of Haidinger's brushes. Spectra of lutein in association with phospholipid model membranes at different temperatures are presented and these support the theory that such incorporation occurs. Additional experiments test an alternative model for Haidinger's brushes and show it to be inconsistent with the spectroscopic properties of lutein.