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  • Title: A routine high-performance liquid chromatography method for carotenoid determination in ultrafrozen orange juices.
    Author: Meléndez-Martínez AJ, Vicario IM, Heredia FJ.
    Journal: J Agric Food Chem; 2003 Jul 16; 51(15):4219-24. PubMed ID: 12848488.
    Abstract:
    An isocratic reversed-phase high-performance liquid chromatography method was developed for routine analysis of the main carotenoids related to the color of orange juice, using a more selective wavelength (486 nm) in which the absorption in the red-orange region of the visible spectra is maximum. Separation was carried out using as the mobile phase the mixture methanol:acetonitrile:methylene chloride:water (50:30:15:5, v/v/v/v), to which small amounts of butylated hydroxytoluene and triethylamine were added (0.1%). Identification was made by comparison either with standards obtained by thin-layer chromatography or with spectral data previously reported. The reproducibility of the method was remarkable; coefficients of variation for the most polar xanthophylls were under 1 and 4% for retention times and areas, respectively. Its application to Valencia late ultrafrozen orange juices has shown that major carotenoids are lutein + zeaxanthin (36%), lutein 5,6-epoxide (16%), antheraxanthin (14%), and beta-cryptoxanthin (12%).
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