Thirteen geometric isomers of retinol, retinal and retinal oxime were resolved in a single chromatographic run on two 250 X 4 mm 5-micron LiChrosorb Si 60 columns in series, using 11.2% ethyl acetate, 1.4% 1-octanol and 2% dioxane in hexane as the mobile phase. All the 11-cis and all-trans isomers of retinol and retinal oxime were completely resolved from each other and from the 9-cis and 13-cis isomers. This chromatographic procedure should be particularly useful for quantifying geometric isomers of retinoids in methylene chloride-hydroxylamine extracts of biological samples such as ocular tissues.