Introduction

1. PC2 PC2 Study Design PC2 Intervention day 1 Intervention day 2 Intervention day 3 Blood & urine samples at: Blood & urine samples at: Blood & urine samples at: * ° * ° * ° † † † 0 2 4 6 hours 0 2 4 6 hours 0 2 4 6 hours PC1 PC1 PC1 * Consumption of 60 g chocolate and 400 mL Water ° Consumption of 200 mL Water † Consumption of 200 mL Water SDC N = 7 Enriched Dark Choc Dark Choc White Choc WC N = 7 Enriched Dark Choc White Choc Dark Choc DCR N = 7 White Choc Enriched Dark Choc Dark Choc Dark Choc SDC N = 7 White Choc Enriched Dark Choc N = 7 Dark Choc White Choc Enriched Dark Choc N = 7 Enriched Dark Choc White Choc Dark Choc S = 42 Washout 1 Washout 2 14 days 14 days Enriched Dark Chocolate: ~900 mg flavan-3-ols and procyanidins Dark Chocolate: ~400 mg flavan-3-ols and procyanidins White Chocolate: NO flavan-3-ols and procyanidins 600 MHz NMR spectrometer with cryoprobe 368 urine samples randomised 1H NMR spectra ppm Sorting and bucketing processing Proton NMR reveals effect on urine metabolite profiles after consumption of polyphenol enriched dark chocolate Luisa M. Ostertag1, Henri Tapp2 Garry G. Duthie1, Ian Colquhoun2, Baukje de Roos1, Kate Kemsley2, Paul A. Kroon2, Gwénaëlle Le Gall2 1University of Aberdeen, Rowett Institute of Nutrition & Health, Aberdeen, UK2 Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA, UK gwenaelle.legall@bbsrc.ac.uk Results Introduction • Cardiovascular disease (CVD) is a primary cause of premature deaths in UK and Worldwide • Activated platelets contribute to early inflammatory events in atherogenesis, which is an important mechanism in the development of CVD • Principal Component Analysis (PCA) shows a clear separation between time 0, 2h and 6 hours post-ingestion • Dietary components such ad polyphenols, which are present in cocoa, fruits, vegetables, herbs, spices, teas and wines may effectively prevent platelet activation thus may aid the prevention of CVD • A set-up randomised controlled cross-over intervention trial was assessed by 1H NMR spectroscopy to evaluate the acute effects of consumption of flavonoid-enriched dark chocolate, compared with standard dark chocolate and white chocolate on the urinary profiles of 42 volunteers Figure 1. PCA on the 1H NMR urine spectra at time 0 and post-ingestion of flavonoid-enriched dark chocolate, standard dark chocolate and white chocolate. Median trajectories (n = 42 volunteers) are similar on PC1 while PC2 levels the effects of the dietary treatments with the largest effect being observed for the flavonoid-enriched dark chocolate Data Recording Figure 2. 1H NMR spectra of volunteer 5 at time 6 hours (flavonoid-enriched dark chocolate, standard dark chocolate and white chocolate). Key to legend: A, alkaloid compounds; B, polyphenols; 7-me-xan, 7-methyl-xanthine; hipp, hippurate; gly, glcycine Conclusions • These results demonstrate the power of untargeted NMR-based metabolite profiling to reveal the modulation of human metabolism following a controlled dietary challenge • We hope to help decipher the modulation induced by polyphenol-rich chocolate by identifying the dietary and endogenous marker metabolites in the 1H NMR spectra We thank the British Biotechnological Science Research Council (BBSRC), the Scottish Government Rural and Environment Research and Analysis Directorate (RERAD), and Natraceutical Industrial S. L. U.