Simultaneous high-performance liquid chromatographic analysis of free carotenoids and carotenoid esters

High-performance liquid chromatography using a non-aqueous reversed phase with gradient elution on C18 columns is a powerful tool for investigating the carotenoid composition of natural samples, e.g., flower petals, and for the simultaneous detection of carotenoids of the widest possible polarity range (xanthophylls, diones, hydrocarbons and carotenoid esters). The comparison of sample extracts submitted or not to saponification allows the presence of carotenoid esters to be revealed through the appearance of the corresponding free hydroxycarotenoids. The gas chromatographic analysis of the fatty acids after alkaline hydrolysis of esters provides further confirmation. In most cases, peaks of various carotenoids were identified by comparison with standards. The wavelengths of the visible absorbance maxima of the chromatographed carotenoids as obtained on-line by the stop-flow method in the eluent system and off-line in carbon disulphide are reported. The esters appear to constitute the main carotenoid fraction in flower petals.