A reversed-phase high-performance liquid chromatographic method to analyze retinal isomers.
A high-performance liquid chromatographic (HPLC) procedure was developed to separate all-trans-, 13-cis-, 11-cis- and 9-cis-retinal isomers. Two reversed-phase Vydac C18 columns in series were used with an isocratic solvent system of 0.1 M ammonium acetate-acetonitrile (40:60, v/v) as mobile phase and all-trans-9-(4-methoxy-2,3,6-trimethylphenyl)-3,7-dimethyl-2,4,6,8-no natetraene-1-ol (TMMP) as internal standard. Prior to HPLC, the retinal isomers were efficiently extracted in their original isomeric conformation using dichloromethane-n-hexane in the presence of formaldehyde. This technique is suitable for the assay of 11-cis- and all-trans-retinal isomers in retina.
Cia
D
D
Laboratoire de Biophysique Sensorielle, Facultés de Médecine et de Pharmacie, Clermont-Ferrand, France. david.cia@u-clermont1.fr
Bonhomme
B
B
Azim
M
M
Wada
A
A
Doly
M
M
Azaïs-Braesco
W
W
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D007202:Indicators and Reagents; D007536:Isomerism; D008968:Molecular Conformation; D011725:Pyridines; D051381:Rats; D012015:Reference Standards; D012160:Retina / Q000737:chemistry; D012172:Retinaldehyde / Q000032:analysis*; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet
D007202:Indicators and Reagents; D011725:Pyridines; D012997:Solvents; C053572:1-methyl-4-(methylpyrrol-2-yl)-1,2,3,6-tetrahydropyridine; D012172:Retinaldehyde