Physical activity increases the bioavailability of flavanones after dietary aronia-citrus juice intake in triathletes S. Medina a , R. Domínguez-Perles a , C. García-Viguera a , R. Cejuela-Anta b , J.M. Martínez-Sanz b , F. Ferreres a , A. Gil-Izquierdo a,⇑ a Department of Food Science and Technology, CEBAS-CSIC, P.O. Box 164, Espinardo, 30100, Murcia, Spain b Department of Physical Education and Sport, Faculty of Education, University of Alicante, Campus de San Vicent del Raspeig, 03540 San Vicent del Raspeig, Alicante, Spain article info Article history: Received 16 May 2012 Received in revised form 25 June 2012 Accepted 12 July 2012 Available online 25 July 2012 Keywords: Aronia-citrus juice Bioavailability Dietary intervention Flavanones Triathletes abstract Control and triathlete volunteers (n = 8 and n = 15, respectively) were given 400 mL and 200 mL of aro- nia-citrus juice (AC-juice), respectively. The 24 h urine samples were hydrolysed to determine the flava- nones concentration by UPLC-QqQ-MS/MS. The flavanones metabolites in both groups of volunteers were glucuronides, sulfates, and sulfo-glucuronides, and the total excretion of flavanones increased fivefold in the triathletes compared with the control volunteers. The increase of ninefold in the homoeriodictyol of triathletes compared to control volunteers may suggest the overactivation of the microbiota metabolism caused by physical exercise. No differences concerning the bioavailability were detected between men and women in controlboth groups. The AC-juice could provide synergistic effects on health due to the increase in the bioavailability of flavanones, avoiding the deleterious effects caused by the overdosage of nutritional supplements. Ó 2012 Elsevier Ltd. All rights reserved. 1. Introduction In the past decade, a number of clinical trials based on dietary interventions have been performed to establish the bioefficiency of distinct subclasses of polyphenols (Kay, 2010). The applicability of polyphenols to the athlete’s world and their health benefits re- main scarcely addressed (Trombold, Barnes, Critchley, & Coyle, 2010). In order to gain a further insight into the relationship be- tween training, nutrition, and health, a variety of nutritional sup- plements have been developed to increase the physical outcome of the training programs regardless of the natural option of fruit juices with bioactive components (Trombold et al., 2010). Citrus juices are known for their high content in flavonoids, especially flavanones (400–600 mg L 1 ) (Gil-Izquierdo, Gil, & Ferreres, 2002; Gil-Izquierdo, Gil, Ferreres, & Tomás-Barberán, 2001). These compounds are mostly attached to rhamnoglucosides which need to be removed by the colon microflora in order to be absorbed (Silberberg et al., 2006). Flavanones have shown a more permanent systemic level, due to the enterohepatic cycle, which allows the re-excretion of metabolites, by bile, and their reabsorp- tion in the small intestine or colon, and therefore, a longer resi- dency at physiological level (Manach, Morand, Gil-Izquierdo, Bouteloup-Demange, & Rémésy, 2003). In recent years, research in this field has focused on the aug- mentation of flavanones bioavailability, by different mechanisms, in order to increase the health-promoting properties of citrus juices. The combination of aronia (Aronia melanocarpa) with citrus juices could provide synergistic effects of flavanones plus anthocy- anins, among other bioactive compounds (Habauzit et al., 2011). However, as far as we are aware, the effect of physical activity on the bioavailability of the target compounds (flavanones) from aro- nia-citrus juices (AC-juice) remains unknown. The aim of the present study was to identify the circulating flav- anones metabolites after the intake of AC-juice, and compare their bioavailability in triathletes with that found in control volunteers. In addition, the flavanones excretion was also evaluated, for a week before and after AC-juice intake, in triathletes. 2. Methods and materials 2.1. Chemicals Naringenin, eriodictyol, homoeriodictyol, hesperetin, and iso- sakuranetin were purchased from Extrasynthèse (Genay, France). Hesperetin 7-O-glucuronide was synthesised in our lab according to the method described by Boumendjel, Blanc, Williamson, and Barron (2009). All LC–MS grade solvents were obtained from J.T. Baker (Phillipsburg, NJ). Formic acid and chlorhydric acid were purchased from Panreac (Barcelona, Spain). The b-glucuronidase, 0308-8146/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.foodchem.2012.07.080 ⇑ Corresponding author. Tel.: +34 968 396363; fax: +34 968 396213. E-mail address: angelgil@cebas.csic.es (A. Gil-Izquierdo). Food Chemistry 135 (2012) 2133–2137 Contents lists available at SciVerse ScienceDirect Food Chemistry journal homepage: www.elsevier.com/locate/foodchem