Abstract

This study was undertaken to investigate the distribution of vitamin E in the macular and peripheral regions of the human retina, retinal pigment epithelium (RPE) and choroid as a function of age. High-performance liquid chromatography (HPLC) was used to measure alpha- and gamma-tocopherol levels quantitatively using tocol as an internal standard. In 57 out of 70 donor eyes (ages 9-104) the macular region was isolated and the tocopherols analyzed. The conventional brush method and a new vortex method were used to isolate the retinal pigment epithelium cells. Similar trends for the vitamin E levels (increase to the 5th decade, decrease after 7th decade) were found for the macular and peripheral retina and for the macular RPE. In the peripheral RPE a slight continuous increase with age was found. The vitamin E levels are higher in the RPE than in the retina, for both macular and peripheral regions. The amounts of vitamin E/mg protein are lower in the macular retina than in the peripheral retina, whereas in the RPE there is no difference in vitamin E content between macular and peripheral regions. A simple method based on a gentle vortex step was found to offer several advantages over the more generally used isolation of RPE cells based on brushing, and there was no difference in recovery of vitamin E in RPE cells when they were isolated by either isolation technique. It was also found that denominators, used to express the values of vitamin E in tissues should, be used with care since age dependent trends in parameters/denominators could be caused by trends in the denominators only.