HPLC analysis of the retinoid content of eyes of Rpe65−/−/Nrl−/− mice. (A) HPLC traces of total retinoid extracts from two eyes of a Rpe65−/−/Nrl−/− mouse. According to retention times and absorbance spectra, the peaks were assigned as follows (arrows): 1, retinyl esters; 2, syn 11-cis retinal oxime; 3, syn all-trans retinal oxime; 4, anti-11-cis retinal oxime; 5, 11-cis retinol; 6, all-trans retinol. (B) HPLC traces of the retinyl ester fraction collected (as denoted with a bar over the peak on trace a), and after saponification and rechromatography (trace b). Inset: Peak absorbance spectra for the saponification product on trace b and the standard compounds 11-cis retinol (trace c, peak 1) and all-trans retinol (trace c, peak 2). (C) Retinoid content of eyes from Rpe65−/−/Nrl−/−, Nrl−/−, Rpe65−/−, and wild-type mice.