Determination of lycopene, alpha- and beta-carotene and retinyl esters in human serum by reversed-phase high performance liquid chromatography.

A rapid specific, microdetermination of the major human blood carotenoids by high performance liquid chromatography (HPLC) separation and quantitation at 466 nm is detailed in this paper. Serum retinyl esters can also be quantified utilizing the same separation procedure but detected at 325 nm. One hundred microliters of deproteinated serum were extracted with chloroform and injected on a reverse-phase column. Separation occurred within 16 min for all compounds of interest employing a mobile solvent of MeOH/AcN/CHCl3 (47:47:6). All compounds were quantified at the wavelengths cited by integrated peak areas using retinyl acetate as a daily standard. Analysis of serum from a hypercarotenemic anorexia nervosa patient and a person suffering from hypervitaminosis A are presented as examples of the clinical application of this procedure.