Uncertainty of antioxidant profiling in complex mixtures using liquid chromatography involving post-column derivatisation |
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| Institution: | 1. Department of Analytical Chemistry, Institute of Chemistry, Faculty of Science, Siedlce University of Natural Sciences and Humanities, 3 Maja 54, 08-110 Siedlce, Poland;2. Department of Chemical and Process Engineering, Chemical Faculty, Gdańsk University of Technology, 11/12 Narutowicza St, 80-233 Gdańsk, Poland;3. Section of Toxicology and Bioanalytics, Department of Civil and Environmental Engineering, Koszalin University of Technology, ?niadeckich 2, 75-453 Koszalin, Poland;4. Polish Society of the Separation Science, Poland;1. Laboratorio de Biofisicoquímica, Departamento de Fisicoquímica, Facultad de Química, Universidad Nacional Autónoma de México, México D. F. 04510, Mexico;2. Facultad de Farmacia, Universidad Autónoma del Estado de Morelos, Mexico;3. Departamento de Ingeniería Química, Conjunto E, Edificio de Ingeniería Química, Facultad de Química, Universidad Nacional Autónoma de México, Mexico;1. Key Laboratory of Cryogenics, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, P.O. Box 2711, Beijing 100190, China;2. University of Chinese Academy of Sciences, Beijing 100039, China;1. CanAm Bioresearch Inc., Winnipeg, MB R3T 0P4 Canada;2. Department of Chemistry, Wayne State University Detroit, Rm 185 Chemistry, MI 48202, USA;3. Phenomenex Inc., Torrance, CA 90501, USA |
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| Abstract: | The main goal of this paper is to discuss the problems associated with antioxidant profiling in complex samples using a high-throughput HPLC system coupled with post-column derivatisation reactor. Based on the experimental data reported in the literature, we demonstrated that improper optimisation of temperature and/or pH assay conditions performed using an on-line derivatisation reactor may substantially change the antioxidant peaks ratio of targeted phytochemical compounds. It has been found that despite the relatively high stability of flavonoids at high temperature and under binary mobile phase HPLC conditions, the reaction of target compounds with common ABTS, FCR and DPPH radicals at elevated regions (particularly above 100 °C) dramatically changes their antioxidant activity values expressed, for example, as TEAC (Trolox equivalent antioxidant capacity) parameters. In principle, separation and detection processes of antioxidant profiling assay must not significantly affect the antioxidant activity of target compounds. In the case of foods eaten by humans or animals, critical experimental parameters such as pH and temperature concerning interaction of target analytes with derivatisation reagent should be as close as possible to the real physiological values. |
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| Keywords: | Total antioxidant activity Antioxidant fingerprints Trolox equivalent antioxidant capacity Post-column derivatisation HPLC Temperature Food analysis Food composition Analytical method uncertainty |
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