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Michael Vega, Elizabeth R Gaillard; Identifying Novel Fluorophores in Human RPE Melanolipofuscin. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1740.
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© 2017 Association for Research in Vision and Ophthalmology.
To identify novel fluorophores in human RPE melanolipofuscin. Identifying novel fluorophores in melanolipofuscin extracts may lead to the development of new diagnostic techniques for the early detection of age related macular degeneration.
Human RPE melanolipofuscin is extracted from human donor eyes as previously described by Feeney-Burns. Folch extraction is performed to obtain the organic soluble portion. The organic soluble melanolipofuscin is collected, dried under argon, and reconstituted in HPLC grade methanol for use in high performance liquid chromatography tandem mass spectrometry (LC/MS/MS) coupled to a fluorescent detector (Surveyor LC with PDA, Thermo Finnigan LCQ Advantage MS, Surveyor FL). Fluorescence detection and tandem mass spectrometry data are analyzed for the identification and structure elucidation of the fluorescent components of human RPE melanolipofuscin.
Melanolipofuscin extracts from human donor tissue have been subjected to LC/MS/MS. The fluorophore, A2E, has been observed as a component of human RPE melanolipofuscin. The presence of additional fluorophores has been confirmed. Tandem mass spectrometry analysis of these unidentified fluorophores has provided structural information for these vitamin A derivatives.
Human RPE melanolipofuscin is observed to accumulate in the RPE with age and this accumulation has been suggested to closely correlate with the onset of AMD. Fluorescent components of melanolipofuscin have been identified in human melanolipofuscin extracts. These fluorophores may lead to the development of a new fluorescence based diagnostic technique for the early detection of AMD.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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