Interactions between cocoa flavanols and inorganic nitrate: additive effects on endothelial function at achievable dietary amounts

Free Radic Biol Med. 2015 Mar;80:121-8. doi: 10.1016/j.freeradbiomed.2014.12.009. Epub 2014 Dec 18.


Dietary intervention studies have shown that flavanols and inorganic nitrate can improve vascular function, suggesting that these two bioactives may be responsible for beneficial health effects of diets rich in fruits and vegetables. We aimed to study interactions between cocoa flavanols (CF) and nitrate, focusing on absorption, bioavailability, excretion, and efficacy to increase endothelial function. In a double-blind randomized, dose-response crossover study, flow-mediated dilation (FMD) was measured in 15 healthy subjects before and at 1, 2, 3, and 4 h after consumption of CF (1.4-10.9 mg/kg bw) or nitrate (0.1-10 mg/kg bw). To study flavanol-nitrate interactions, an additional intervention trial was performed with nitrate and CF taken in sequence at low and high amounts. FMD was measured before (0 h) and at 1h after ingestion of nitrate (3 or 8.5 mg/kg bw) or water. Then subjects received a CF drink (2.7 or 10.9 mg/kg bw) or a micro- and macronutrient-matched CF-free drink. FMD was measured at 1, 2, and 4 h thereafter. Blood and urine samples were collected and assessed for CF and nitric oxide (NO) metabolites with HPLC and gas-phase reductive chemiluminescence. Finally, intragastric formation of NO after CF and nitrate consumption was investigated. Both CF and nitrate induced similar intake-dependent increases in FMD. Maximal values were achieved at 1 h postingestion and gradually decreased to reach baseline values at 4 h. These effects were additive at low intake levels, whereas CF did not further increase FMD after high nitrate intake. Nitrate did not affect flavanol absorption, bioavailability, or excretion, but CF enhanced nitrate-related gastric NO formation and attenuated the increase in plasma nitrite after nitrate intake. Both flavanols and inorganic nitrate can improve endothelial function in healthy subjects at intake amounts that are achievable with a normal diet. Even low dietary intake of these bioactives may exert relevant effects on endothelial function when ingested together.

Keywords: Bioavailability; Cocoa flavanols; Endothelial function; Free radicals; Interactions; NO; Nitrate.

Publication types

  • Clinical Study
  • Randomized Controlled Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Blood Pressure / drug effects*
  • Blood Pressure / physiology
  • Brachial Artery / drug effects
  • Brachial Artery / physiology
  • Cacao / chemistry*
  • Chromatography, High Pressure Liquid
  • Cross-Over Studies
  • Dietary Supplements*
  • Dose-Response Relationship, Drug
  • Flavonoids / administration & dosage*
  • Flavonoids / blood
  • Flavonoids / urine
  • Gastric Mucosa / metabolism
  • Humans
  • Male
  • Nitrates / administration & dosage*
  • Nitrates / blood
  • Nitrates / urine
  • Nitric Oxide / analysis
  • Nitric Oxide / metabolism
  • Vasodilation / drug effects*
  • Vasodilation / physiology


  • Flavonoids
  • Nitrates
  • Nitric Oxide