Journal of Pharmaceutical and Biomedical Analysis 100 (2014) 348–356 Contents lists available at ScienceDirect Journal of Pharmaceutical and Biomedical Analysis j o ur nal ho me page: www.elsevier.com/lo cate/jpba Changes in urinary metabolic profile after oral administration of curcuma extract in rats Stefano Dall’Acqua a,∗ , Matteo Stocchero b , Maria Clauser a , Irene Boschiero a , Emmanuel Ndoum c , Mariano Schiavon a , Stefano Mammi c , Elisabetta Schievano c a Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Via Marzolo 5, 35131 Padova, Italy b S-IN Soluzioni Informatiche, Via Ferrari 14, 36100 Vicenza, Italy c Department of Chemical Sciences, University of Padova, Via Marzolo 1, 35131 Padova, Italy a r t i c l e i n f o Article history: Received 26 April 2014 Received in revised form 23 July 2014 Accepted 28 July 2014 Available online 12 August 2014 Keywords: Curcumin Metabolomic NMR HPLC-MS ASCA a b s t r a c t The diffusion of phytochemicals in health promoting products is growing, but studies related to their effects on healthy subjects are still lacking despite the large consumption of natural products as nutraceuticals or food supplements. In many cases, research supports the in vitro antioxidant activity of phytochemicals, but the health claims attributed to the final marketed nutraceutical products have dubious scientific foundation. Also, studies focussed on the definition of their biological targets and mechanisms of action can be useful to assess their efficacy and safety. In this study, the effect of oral administration of 80 mg/kg of Curcuma longa Linn. extract to 12 healthy rats over 25 days was evaluated by monitoring the changes of urinary composition. 24-h urine was collected during the animal experiment and the composition was analyzed by 1 H NMR and HPLC–MS. The two datasets were studied individually through a metabolomic approach and the multivariate analysis revealed significant differences between the control and the treated group. Curcumin levels were also measured in 24-h urine samples by HPLC–MS. Both the 1 H NMR and the HPLC–MS dataset showed that the administration of 80 mg/kg of Curcuma longa extract to healthy animals induces changes in urinary composition. Decreased allantoin urinary levels can be considered a partial demonstration of the in vivo effect of curcumin on oxidative stress in a healthy animal model. © 2014 Elsevier B.V. All rights reserved. 1. Introduction Many epidemiological studies have shown the relationship between dietary habits and a reduced disease risk indicating the strong impact of food and nutrition on health. The consumption of plant-derived foods is correlated to health improvement and to the prevention of several degenerative diseases, especially age-related ones. There is an emerging use of so-called “nutraceuticals” due to their supposed health promotion role. Such products are not therapeutics and are mainly used by healthy people at low doses and for relatively long periods of time [1–3]. To the best of our knowledge, studies on long-term supplementation to evaluate the biological effects after regular intake of these supplements, are gen- erally missing. The ability of such plant-derived foods to reduce the risk to develop chronic diseases has been associated to the occurrence of phytochemicals such as polyphenols [4], investigated ∗ Corresponding author. Tel.: +39 049 8275344; fax: +39 049 8275366. E-mail address: stefano.dallacqua@unipd.it (S. Dall’Acqua). for many years especially for their antioxidant properties [1,4,5]. Although health benefits of many plant-derived polyphenols are ascribed to their antioxidant properties in vitro, evidence for in vivo antioxidant effects is limited: no validated in vivo biomarkers have been identified, no long-term studies are available, and knowledge of bioavailability is missing. The apparent discrepancy between in vitro and in vivo studies may be due to the poor plasma concentra- tion and extensive metabolism of these molecules [6,7]. The pathogenesis of many diseases is rather multi-factorial and not due to a single cause; thus, multi-targeted antioxidant foods could be important in the area of preventing diseases. If the effi- cacy of nutraceuticals and plant-derived antioxidants is based on the combined action of different constituents acting with multi- targeted mechanisms, a holistic approach to study the efficacy of phytochemicals is necessary. “-Omics” technologies may be very useful to explain biological effects of different antioxidant supple- ments [8]. Because phytochemicals introduced with the diet or with supplementation act on the whole metabolome, the evaluation of the entire metabolic changes induced by the phytoconstituents and http://dx.doi.org/10.1016/j.jpba.2014.07.035 0731-7085/© 2014 Elsevier B.V. All rights reserved.