28] High-pressure liquid chromatography of vitamin A metabolites and analogs

Abstract

Publisher Summary This chapter discusses high-pressure liquid chromatography (HPLC) of vitamin A metabolites and analogs. The application of HPLC to the separation of retinoids has revolutionized vitamin A chromatography and has proved to be an extremely powerful and valuable method for studying vitamin A metabolism. The use of HPLC offers several advantages over conventional methods of vitamin A chromatography, including rapid separation, excellent resolution, quantitative recovery, elimination of artifacts, and the capability of chromatographing a wide range of vitamin A compounds with only minor manipulation of the chromatographic solvent. Straight-phase HPLC on microparticulate silica gel columns has been applied to the study of retinol storage and distribution, retinal isomers and analogs, determination of serum retinol and retinoic acid, and isolation of urinary and fecal metabolites of retinoic acid. This chapter describes the high-pressure liquid chromatography methods developed and used in the authors' laboratory for separation of vitamin A metabolites and analogs. Straight-phase and reverse-phase high-pressure liquid chromatography methods are presented and their application to biochemical problems in the vitamin A field are discussed.