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The ability to measure endogenous metabolites of retinoids (vitamin A and its derivatives) in biological samples is key tounderstanding the crucial physiological actions of vitamin A. Over the years, many assays and methods have been developedto analyze different retinoids in biological samples. Liquid chromatography is the best analytical technique for routine analysisof these compounds. However, due to their different chemical properties, different retinoid metabolites cannot be accuratelyseparated and quantified in a single chromatographic run. Here, we will describe a reverse-phase HPLC isocratic method thatenables extraction, separation, identification, and quantification of all-
trans
-retinol and different molecular species of retinyl ester with high accuracy, sensitivity, and reliability.