title: Effects of organic solvents on two retinal pigment epithelial lipofuscin fluorophores, A2E and all-trans-retinal dimer.
abstract: Gene and drug therapies are being developed to alleviate vision loss in patients with Stargardt's disease and age-related macular degeneration (AMD). To evaluate the therapeutic effects of these treatments, organic solvents are routinely used to extract and quantify bisretinoid lipofuscin constituents, such as N-retinylidene-N-retinyl-ethanolamine (A2E) and all-trans-retinal dimer (ATR-dimer). By high-performance liquid chromatography (HPLC), we found that A2E and ATR-dimer were both altered by tetrahydrofuran (THF) and chloroform, but were stable in dimethyl sulfoxide (DMSO) or methanol (MeOH). In addition, cyclohexane and ethanol (EtOH) did not alter ATR-dimer, whereas an alteration of A2E occurred in EtOH. On the basis of these findings, we designed processes II-IV, generated by modifications of process I, a routine method to measure bisretinoid compounds in vivo. Extra amounts of either ATR-dimer or A2E in mouse eyecups were released by processes II-IV versus process I. Efforts to clarify the effects of organic solvents on lipofuscin pigments are important because such studies can guide the handling of these fluorophores in related experiments.
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China; Eye Center, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China; School of Medicine, Zhejiang University City College, Hangzhou 310015, China.
authors.forename: Qiu-xia
authors.initials: QX
authors.lastname: Jin
authors.forename: Xin-ran
authors.initials: XR
authors.lastname: Dong
authors.forename: Jing-meng
authors.initials: JM
authors.lastname: Chen
authors.forename: Ke
authors.initials: K
authors.lastname: Yao
authors.forename: Ya-lin
authors.initials: YL
authors.lastname: Wu
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D008062:Lipofuscin / Q000032:analysis*; D008268:Macular Degeneration / Q000628:therapy; D051379:Mice; D008810:Mice, Inbred C57BL; D010857:Pigment Epithelium of Eye / Q000737:chemistry*; D012172:Retinaldehyde / Q000031:analogs & derivatives* / Q000032:analysis; D012997:Solvents; D000080362:Stargardt Disease
chemical_list: D008062:Lipofuscin; D012997:Solvents; C525876:retinal dimer; D012172:Retinaldehyde
keywords: All-trans-retinal dimer; Lipofuscin; N-retinylidene-N-retinyl-ethanolamine (A2E); Organic solvents; Retinal pigment epithelial