Carotenoids high pressure liquid chromatography

For many decades, the standard technique for measuring carotenoids has been high-pressure liquid chromatography (HPLC). This time consuming and expensive chemical method works well for the measurement of carotenoids in serum, but it is difficult to perform in human tissue since it requires biopsies of relatively large tissue volumes. Additionally, serum antioxidant measurements are more indicative of short-term dietary intakes of antioxidants rather than steady-state accumulations in body tissues exposed to external oxidative stress factors such as smoking and UV-light exposure. 

The most commonly used method for the identification of carotenoids is high-pressure liquid chromatography (HPFC) combined with the UV-Vis absorption detection. The introduction of diode array detection enabled parallel collection of pigment spectra, which greatly aids the quantification and localization of unknown compounds. Coupling HPFC with the mass-spectrometer significantly... 

The development of ultra high-pressure liquid chromatography technology has dramatically reduced the analysis time for a number of small molecules. However, we have found that the analysis of carotenes is poor with the currently available column chemistries. The development of C30 columns with backbones that can withstand ultra high pressure would allow carotenoid chemists to take full advantage of this technology. 

As the last physical method useful for stereochemical studies of carotenoids one should mention high pressure liquid chromatography. This was first applied to the carotenoid field in 1971 by Stewart and Wheaton 159) and subsequently shown to be superior for separation of geometrical isomers (75, 76, 158) and diastereomers (46, 75). Future developments on a preparative scale are anticipated. 

Fiksdahl, a., J. T. Mortensen, and S. Liaaen-Jensen High-pressure liquid chromatography of carotenoids. J. Chromatography 157, 111 (1978). 

Lacker T, Strohschein S and Albert K, Separation and identification of various carotenoids by C30 reversed-phase high-performance liquid chromatography coupled to UV and atmospheric pressure chemical ionization mass spectrometric detection. J Chromatogr A 854 37 14 (1999). 

Clarke, P.A. Barnes, K.A. Startin, J.R. Ibe, F.L Shepherd, M.J. 1996. High-performance liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry for the determination of carotenoids. Rapid Comm. Mass Spectrom. 10 1781-1785. 

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