CHROMATOGRAPHY OF RETINOIDS

This article reviews the determination of retinoic acids and their metabolites (first-generation retinoids), aromatic retinoids (second generation) and arotinoids (third generation) in biological samples. Because of the sensitivity of the retinoids to isomerization and oxidation, special care has to be taken from sample collection and storage, throughout extraction, till the final chromatographic separation. High and strong protein binding, and insolubility in aqueous solutions hamper the extraction from biological samples. Various extraction procedures are discussed, mainly involving liquid-liquid extraction of biological fluids or lyophilized tissue samples. The new technique involving direct injection of biological fluids or tissue homogenates, using high-performance liquid chromatography (HPLC) with automated column switching, provides full protection from light and simplifies sample work-up. HPLC with ultraviolet detection is the method of choice for the determination of retinoids, because it is rapid, sensitive and allows separation of geometric isomers and metabolites within a wide polarity range. Gas chromatography-mass spectrometry is not appropriate for first- and second-generation retinoids because of isomerization, but allows very sensitive determination of third-generation retinoids, although very extensive sample clean-up and derivatization are necessary. However, direct injection of large volumes of biological fluids into HPLC systems, using on-line solid-phase extraction and automated column-switching, results in very sensitive methods even with simple ultraviolet detection and may become the method of choice for routine analyses. © 1990.