Rapid Folin-Ciocalteu method using microtiter 96-well plate cartridges for solid phase extraction to assess urinary total phenolic compounds, as a biomarker of total polyphenols intake |
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Authors: | Alexander Medina-Remón Ana Barrionuevo-González Raúl Zamora-Ros Cristina Andres-Lacueva Ramón Estruch Miguel-Ángel Martínez-González Javier Diez-Espino |
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Institution: | a Nutrition and Food Science Department, XaRTA, INSA Pharmacy School, University of Barcelona, Av. Joan XXIII s/n, 08028 Barcelona, Spain b Department of Internal Medicine, Hospital Clínic, Institut d’Investigació Biomèdica August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain c CIBER 06/003 Physiopathology of Obesity and Nutrition (CIBEROBN), and RETICS RD06/0045/0003, Instituto de Salud Carlos III, Spain d Ingenio-CONSOLIDER program, FUN-C-FOOD, Instituto de Salud Carlos III, Barcelona, Spain e Department of Preventive Medicine and Public Health, Clínica Universitaria, University of Navarra, Pamplona, Spain f Tafalla Health Center, Osasunbidea, Tafalla, Spain |
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Abstract: | Nutritional markers have several advantages for epidemiologic and clinical assays, when compared to dietary data obtained by food frequency questionnaires. Few studies have assessed whether total polyphenol (TP) compounds provide a valid biomarker for TP intake. To date, there has been almost no literature describing methods to determine TP in complex matrices such as urine, which have many interfering substances.We report a rapid Folin-Ciocalteu method to determine TP in urine samples using Oasis® MAX 96-well plate cartridges for solid phase extraction. These plates allow analysis of a high number of samples at the same time. We performed a prospective, randomized, crossover trial and one cross-sectional study with 60 volunteers from the PREDIMED trial, seeking to evaluate whether the TP in urine were correlated with polyphenol intake and could, therefore, be considered as a marker of intake of these compounds.The assay was optimized; the sensitivity and the polarity range of urine polyphenols were increased and the detection and quantification limits were significantly reduced. The metabolites in standards solution and urine samples were stable under the storage and handling conditions. In the clinical trial and the cross-sectional study, TP excreted in spot urine samples were positively correlated with TP intake, r = 0.48, P < 0.01 and r = 0.257, P = 0.04, respectively.The methodology described may be used to detect TP in urine samples, employing the high throughput of 96-well microtiter plates and reader. The method is fast and simple and it allows analysis of a large number of samples at the same time. |
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Keywords: | Solid phase extraction 96-Well plate Phenols Folin-Ciocalteu Biomarker Urine PREDIMED Polyphenols stability |
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