British Journal of Nutrition

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British Journal of Nutrition (2010), 103:1008-1014 Cambridge University Press
Copyright © The Authors 2009

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Human and Clinical Nutrition

Effect of dark chocolate on plasma epicatechin levels, DNA resistance to oxidative stress and total antioxidant activity in healthy subjects

A. Spadafrancaa1 c1, C. Martinez Conesaa2, S. Sirinia1 and G. Testolina1

a1 Department of Food Science and Microbiology (di.S.T.A.M), Division of Human Nutrition, University of Milan, Via Celoria, 2, 20133 Milan, Italy
a2 Murcian Institute of Investigation and Agricultural Development (IMIDA), La Alberca (Murcia), Spain
Article author query
spadafranca a [PubMed]  [Google Scholar]
martinez conesa c [PubMed]  [Google Scholar]
sirini s [PubMed]  [Google Scholar]
testolin g [PubMed]  [Google Scholar]


Dark chocolate (DC) may be cardioprotective by antioxidant properties of flavonoids. We investigated the effect of DC (860 mg polyphenols, of which 58 mg epicatechin) compared with white chocolate (WC; 5 mg polyphenols, undetectable epicatechin) on plasma epicatechin levels, mononuclear blood cells (MNBC) DNA damage and plasma total antioxidant activity (TAA). Twenty healthy subjects followed a balanced diet (55 % of energy from carbohydrates, 30 % from fat and 1 g protein/kg body weight) for 4 weeks. Since the 14th day until the 27th day, they introduced daily 45 g of either WC (n 10) or DC (n 10). Whole experimental period was standardised in antioxidant intake. Blood samples were collected at T0, after 2 weeks (T14), 2 h and 22 h after the first chocolate intake (T14+2 h and T14+22 h), and at 27th day, before chocolate intake (T27), 2 h and 22 h after (T27+2 h and T27+22 h). Samples, except for T14+2 h and T27+2 h, were fasting collected. Detectable epicatechin levels were observed exclusively 2 h after DC intake (T14+2 h = 0·362 (se 0·052) μmol/l and T27+2 h = 0·369 (se 0·041) μmol/l); at the same times corresponded lower MNBC DNA damages (T14+2 h = − 19·4 (se 3·4) % v. T14, P < 0·05; T27+2 h = − 24 (se 7·4) % v. T27, P < 0·05; T14+2 h v. T27+2 h, P = 0·7). Both effects were no longer evident after 22 h. No effect was observed on TAA. WC did not affect any variable. DC may transiently improve DNA resistance to oxidative stress, probably for flavonoid kinetics.

(Received February 18 2009)

(Revised October 02 2009)

(Accepted October 06 2009)

(Online publication November 05 2009)

Key Words:Dark chocolate; Epicatechin; DNA damage; Total antioxidant activity


c1 Corresponding author: Dr Angela Spadafranca, fax +39 02 503 16 191, email


Abbreviations: DC, dark chocolate; MNBC, mononuclear blood cells; SD, standardised diet; TAA, total antioxidant activity; Trolox, ( ± )-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid; WC, white chocolate