Carotenoids supercritical-fluid chromatography

The use of supercritical fluid chromatography for carotene separation has been examined and optimized, especially in regard to temperature, pressure, and organic modifiers in the supercritical fluid (71). With an RP column it was possible to resolve an a-carotene-cis isomer from an all-trans carotene as well as two cis isomers of /3-carotene from an all-trans /3-carotene. As with HPLC, only polymeric C,8 columns were able to resolve the cis isomers of a- and /3-carotene from the all-trans isomers. Supercritical fluid chromatography offers the advantage not only of an efficient separation but also of fast analysis. Indeed, the use of SFC with ODS-based columns for the analysis of carotenoid pigments affords a threefold reduction of analysis time compared to HPLC (72). The elution order of carotenoids and their cis isomers was found to be the same as in RP-HPLC. The selectivity of the system could further be increased by adding modifiers (e.g.,... 

L. H. Tonucci, G. R. Beecher, A.. P. Emery, and S. N. Chesler, Supercritical Fluid Extraction of Carotenoids from Foods, Proc. 4th International Symposium on Supercritical Fluid Chromatography and Extraction, Cincinnati, Ohio, USA, pp. 119-120, May 1992. 

D. R. Gere, Separation of Paprika Oleoresins and Associated Carotenoids by Supercritical Fluid Chromatography, Hewlett-Packard AppHcation Note AN 800-5, Avondale, Pennsylvania, USA, (1983). 

Silver ion chromatography is used primarily in liquid [213,215-218], thin-layer [212,213,216,219], and supercritical fluid chromatography [220-222] for the separation of fatty acid derivatives and triacylglycerols, and to a lesser extent, terpenes, sterols, carotenoids, and pheromones. Silver ion chromatography is widely used on its own, or as a preliminary simplification step, to elucidate the stmctures of fats, oils and lipids [213]. It is possible to fractionate animal or fish oils into fractions with zero to six double bonds. Species with the same total number of double bonds can be separated based on the difference in the number of double bonds in individual acyl residues. 

Supercritical Fluid Chromatography-Mass Spectrometry of Carotenoid Pigments... 

E Lesellier, A Tchapla, C Marty, A Lebert. Analysis of carotenoids by high-performance liquid chromatography and supercritical fluid chromatography. J Chromatogr 633 9-23, 1993. 

H Pfander, U NiggU. Chromatography Part V. Supercritical-fluid chromatography. In G Britton, S Liaaen-Jensen, H Pfander, eds. Carotenoids. lA Isolation and Analysis. Basel Birkhauser Verlag, 1995, pp 191-198. 

Matsubara, A., Uchikata, T., Shinohara, M., Nishiumi, S., Yoshida, M., Fukusaki, E. Bamba, T. (2012). Highly sensitive and rapid profiling method for carotenoids and their epoxidized products using supercritical fluid chromatography coupled with electrospray ionization-triple quadrapole mass speetrometry. J. Biosci Bioeng., 113, 782-787. 

High performance liquid chromatography (HPLC) has been by far the most important method for separating chlorophylls. Open column chromatography and thin layer chromatography are still used for clean-up procedures to isolate and separate carotenoids and other lipids from chlorophylls and for preparative applications, but both are losing importance for analytical purposes due to their low resolution and have been replaced by more effective techniques like solid phase, supercritical fluid extraction and counter current chromatography. The whole analysis should be as brief as possible, since each additional step is a potential source of epimers and allomers. 

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