On-line coupling of automatic solid-phase extraction and HPLC for determination of carotenoids in serum |
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Authors: | Ferreiro-Vera C Mata-Granados J M Quesada Gómez J M Luque de Castro M D |
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Institution: | a Department of Analytical Chemistry, Annex C-3, Campus of Rabanales, University of Córdoba, E-14071, Córdoba, Spain b Institute of Biomedical Research Maimónides (IMIBIC), Reina Sofía Hospital, University of Córdoba, E-14071, Córdoba, Spain c Department of I+D+i, Sanyres Group, E-14012, Córdoba, Spain d Mineral Metabolism Unit, Hospital Reina Sofia of Córdoba, E-14004, Córdoba, Spain |
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Abstract: | The automated method developed for the determination of carotenoids uses 200 μL of serum, which was mixed with 400 μL of tetrahydrofuran, vortexed for 1 min, settled for 10 min, centrifuged for 6 min and the supernatant injected into an automatic solid-phase extraction (SPE) system for cleanup-preconcentration. A 10% water-acetonitrile mobile phase at 1.5 mL min−1 eluted the retained compounds and transferred them on-line to a reversed-phase analytical column for individual separation of the target analytes. Visible detection was performed at 450 and 460 nm. The detection limits for the target analytes were between 3 and 30 ng mL−1; the precision (expressed as relative standard deviation) ranged between 2.83 and 5.06% for repeatability and between 3.80 and 7.40% for within laboratory reproducibility. The total analysis time was 18 min. The proposed method is reliable, robust, and has an excellent potential for high-throughput use in both clinical and research laboratories. |
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Keywords: | Carotenoids On-line SPE Serum HPLC-DAD |
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