Acute consumption of three interventions (60 g dark chocolate enriched in flavan-3-ols, 60 g standard dark chocolate, or 60 g white chocolate) on three separate days (at least 2 weeks apart) in random order.
Post-prandial measurements at t = 0 h, t = 2 h and t = 6 h.

Cardiovascular Disease

Acute consumption (within 15 minutes) of 60 g of chocolate containing ~900 mg of total flavan-3-ols and procyanidins.

Dark chocolate enriched in flavan-3-ols and procyanidins

Acute consumption (within 15 minutes) of 60 g of chocolate containing ~400 mg total flavan-3-ols and procyanidins.

Standard dark chocolate

Acute consumption (within 15 minutes) of 60 g of white chocolate containing no flavan-3-ols and procyanidins.

White chocolate

Press release about study to recruit volunteers, 2009-04-23

University of Aberdeen Rowett Institute of Nutrition and Health

Dark Chocolate and Platelet Function in Humans

Acute Effects of the Consumption of Dark Chocolate Enriched in Flavan-3-ols on Platelet Function and the Platelet Proteome

Using a Helena Platelet Aggregation Chromogenic Kinetics System-4 (PACKS-4) light transmission aggregometer
Induced by adenosine diphosphate (ADP) and thrombin receptor-activating peptide (TRAP)

Using collagen-epinephrine coated cartridges.

P-selectin expression as early marker for platelet activation
Activated fibrinogen receptor as late marker for platelet activation
Induced by ADP and TRAP
Using BD FACSArray Bioanalyzer

Using liquid chromatography-tandem mass spectrometry (LC-MS/MS)
Enzyme-hydrolysed for total flavan-3-ols ((-)-epicatechin equivalents)
Non-Hydrolysed for metabolic profile

Using 2D-gel electrophoresis and LC-MS/MS identification of proteins.

Using enzyme-linked immunosorbent assay (ELISA) in plasma after platelet aggregation

Using high performance liquid chromatography (HPLC) and/ or immunoassays

Using a Thermo KONELAB 30 selective chemistry analyser (Thermo Scientific, Hertfordshire, UK) and its respective kit
To be used for normalisation of urinary flavan-3-ols and total phenolics from spot urine samples.

To establish a metabolic profile - markers of intake and potential effects on host metabolism

Plasma levels of lipid peroxides (thiobarbituric acid-reactive substances, TBARS)
Activity of glutathione peroxidase (Only at t = 2 h after chocolate ingestion)

Using the fatty acid methyl ester (FAME) analysis and a gas chromatographic approach

procyanidin d 1/hemorrhage

Dark Chocolate and Platelet Function in Humans

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