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Oxidative Medicine and Cellular Longevity
Volume 2016, Article ID 9268531, 10 pages
Research Article

A New Method to Simultaneously Quantify the Antioxidants: Carotenes, Xanthophylls, and Vitamin A in Human Plasma

1Nutrition and Food Science Department, XaRTA, INSA, Pharmacy School, University of Barcelona, Avenue Joan XXII s/n, 08028 Barcelona, Spain
2CIBER CB06/03 Fisiopatología de la Obesidad y la Nutrición, CIBEROBN, Choupana s/n, 15706 Santiago de Compostela, Spain
3Scientific and Technical Services, University of Barcelona, Baldiri Reixac, 10, 08028 Barcelona, Spain
4Department of Internal Medicine, Hospital Clínic, IDIBAPS, University of Barcelona, Villarroel, 170, 08036 Barcelona, Spain

Received 22 June 2015; Revised 19 August 2015; Accepted 24 August 2015

Academic Editor: Luciano Saso

Copyright © 2016 Mariel Colmán-Martínez et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


A simple and accurate reversed phase high-performance liquid chromatography coupled with diode array detector (HPLC-DAD) method for simultaneously determining and quantifying the antioxidants carotenes, xanthophylls, and retinol in human plasma is presented in this paper. Compounds were extracted with hexane, a C30 column, and a mobile phase of methanol, methyl tert-butyl ether, and water were used for the separation of the compounds. A total of 8 carotenoids, 3 Z--carotene isomers, and 1 fat-soluble vitamin (retinol) were resolved within 72 min at a flow rate of 0.6 mL/min. Detection was achieved at 450 nm for carotenoids and 330 nm for retinol. To evaluate the effectiveness of themethod, it has been applied to an intervention study conducted on eight volunteers. Results. Limits of detection were between 0.1 g/mL for lycopene and astaxanthin and 1.3 g/mL for 15-Z--carotene. Recoveries were ranged between 89% and 113% for -carotene and astaxanthin, respectively. Accuracy was between 90.7% and 112.2% and precision was between 1% and 15% RSD. In human plasma samples compounds studied were identified besides three lycopene isomers, demonstrated to be suitable for application in dietary intervention studies. Conclusions. Due to its accuracy, precision, selectivity, and reproducibility, this method is suitable to dietary habits and/or antioxidants status studies.