Cocoa Group Research Articles

Changes in the flavan-3-ols, anthocyanins, and flavanols composition of cocoa beans of different Theobroma cacao L. groups affected by roasting conditions

In the present study, the effect of roasting conditions on the individual flavan-3-ols, anthocyanins, and flavanols content of cocoa beans of five different Theobroma cacao L. groups was studied. The raw cocoa beans were roasted at four different temperatures (110, 120, 135, and 150 °C) and three different relative air humidities (0.3, 2.0, and 5.0 %). The great variations in the contents of both total and individual phenolic compounds among the different studied cocoa groups were observed. In general, the cocoa beans of the Forastero group from Brazil exhibited the highest content of individual flavan-3-ols, anthocyanins, and flavanols, while the samples of the Trinitario type from Papua New Guinea had the lowest polyphenols levels. Roasting significantly affected the profile and levels of the flavan-3-ols, anthocyanins, and flavanols in cocoa beans. The changes depended upon the different cocoa types evaluated and their processing conditions. A decrease in epicatechin, procyanidin B, procyanidin C1, both anthocyanins, and quercetin glycosides was observed in all of the five cocoa groups tested after roasting. This reduction coincided with the increase in catechin and quercetin contents in roasted cocoa beans from the five cocoa groups.

Assessment of DNA damage using comet assay in middle-aged overweight/obese subjects after following a hypocaloric diet supplemented with cocoa extract.

Nutrient excess and unbalanced diets can result in overproduction of reactive oxygen species (ROS), which are associated with oxidative stress. Cocoa extract contains antioxidants that inhibit the harmful effects of ROS. This trial analysed the effect of cocoa extract consumption integrated as a bioactive compound into ready-to-eat meals, on oxidative stress at the level of DNA in overweight/obese subjects. Fifty volunteers [57.26(5.24) years, 30.59(2.33)kg/m(2)] participated in a 4-week double-blind, randomised, placebo-controlled parallel nutritional intervention. Half of the volunteers received meals supplemented with 1.4 g/day cocoa extract, while the other half received control meals, both within a 15% energy restriction diet. Lymphocytes were isolated and endogenous strand breaks, oxidised bases and resistance to H2O2-induced damage were measured by the comet assay. The intake of ready-to-eat meals supplemented with cocoa extract did not show relevant changes in the oxidative status of DNA. However, in the cocoa group, oxidised bases negatively correlated with methyl epicatechin-O-sulphate (r = -0.76; P = -0.007) and epicatechin sulphate (r = -0.61; P = -0.046). When volunteers of both groups were analysed together, a marginal decrease (P = 0.072) in oxidised bases was observed, which attributed to weight loss. Subjects who started the intervention with higher levels of damage showed a greater reduction in oxidised bases after 4 weeks (P = 0.040) compared to those who had lower baseline levels. In conclusion, even if 1.4 g of cocoa supplementation for 4 weeks did not show notable changes in terms of antioxidant status of DNA, the energy restriction showed a slightly decrease in oxidised bases and this was seen to a greater extent in subjects who started the intervention with higher levels of damage. On the other hand, the inverse associations found between oxidised bases and some cocoa-derived metabolites suggest that a protective effect might be seen in a longer period of time or in subjects with higher baseline DNA damage. www.clinicaltrials.gov (NCT01596309).

Anxiety Behaviour Displayed in C57BL/6J Mice Consuming Coffee and Cocoa, but not Observed in Mice Consuming Japanese Green Tea in a High Fat Diet Induced Obesity Model

It is now acknowledged that certain foods may affect mood and behavior. Recently, the food industry has seen a rise in foods containing beverages such as green tea cake, ice-cream, coffee biscuits as per the traditional chocolate as a food and cocoa beverage. Green tea has been suggested to exert numerous health properties, including anxiolytic properties (Vignes et. al. 2006), as does cocoa (i.e. migrane) (Savi et. al. 2002). Numerous studies have studied the consumption of these beverages (i.e. cocoa, coffee and green tea) for their anti-oxidant effect but few studies have included them in a feed format. In this study, mice were allotted into a 16 week treatment of a high-fat, high carbohydrate diet of varying treatments including 1) control diet (21% fat, 36.1% sucrose), 2) 2% cocoa (21% fat, 36.1% sucrose), 3) 2% coffee (21% fat, 36.1% sucrose) and 4) 2% green tea (21% fat, 36.1% sucrose). Subjects were offered the diets in a non-beverage feed format and fresh diet was offered daily. At week 16, the cocoa group mice displayed quicker entry time to dark areas, a greater (p<0.05) amount of time spent in the dark box area and higher (p<0.05) number of peeking events when compared with the control group. In addition, the coffee group mice displayed a higher (p<0.05) re-entries from the light to the dark box area (i.e. anxiety like behavior/ agitation), whereas the green tea group did not display the same level of anxiety like behavior seen in the cocoa and coffee groups. This result may suggest an anxiolytic offset effect against caffeine in green tea versus coffee or another phytochemical component or body compositional changes when consuming a high fat, high sucrose diet.

Oxidised LDL levels decreases after the consumption of ready-to-eat meals supplemented with cocoa extract within a hypocaloric diet

Background and aimsCocoa flavanols are recognised by their favourable antioxidant and vascular effects. This study investigates the influence on health of the daily consumption of ready-to-eat meals supplemented with cocoa extract within a hypocaloric diet, on middle-aged overweight/obese subjects. Methods and resultsFifty healthy male and female middle-aged volunteers [57.26 ± 5.24 years and body mass index (BMI) 30.59 ± 2.33 kg/m2] were recruited to participate in a 4 week randomised, parallel and double-blind study. After following 3 days on a low-polyphenol diet, 25 volunteers received meals supplemented with 1.4 g of cocoa extract (645.3 mg of polyphenols) and the other 25 participants received control meals, within a 15% energy restriction diet. On the 4th week of intervention individuals in both dietary groups improved (p < 0.05) anthropometric, body composition, blood pressure and blood biochemical measurements. Oxidised LDL cholesterol (oxLDL), showed a higher reduction (p = 0.030) in the cocoa group. Moreover, myeloperoxidase (MPO) levels decreased only in the cocoa supplemented group (p = 0.007). Intercellular Adhesion Molecule-1 (sICAM-1) decreased significantly in both groups, while Vascular Cell Adhesion Molecule-1 (sVCAM-1) did not present differences after the 4 weeks of intervention. Interestingly, cocoa intake showed a different effect by gender, presenting more beneficial effects in men. ConclusionsThe consumption of cocoa extract as part of ready-to-eat meals and within a hypocaloric diet improved oxidative status (oxLDL) in middle-aged subjects, being most remarkable in males.Registration number: Registered at www.clinicaltrials.gov (NCT01596309).

Prevention of diet-induced obesity in C57BL/BJ mice with addition of 2 % dietary green tea but not with cocoa or coffee to a high-fat diet

Most of the fat oxidation effects of green tea have been observed in a beverage form. This study explored the use of green tea as feed formation in obese mice to protect against diet-induced obesity. Forty-eight adult C57BL/6J mice were randomised to either a control diet with 36.1 % w/w sucrose or a standard diet, except for the replacement of 2 % w/w of the sucrose dietary component (36.1 % sucrose in control) with 2 % w/w cocoa (34.1 % sucrose), 2 % w/w coffee, or 2 % w/w powdered Japanese green tea. At 16th week, body composition, plasma glucose, organ samples and bone mineral density were obtained. In comparison to the control group, only the 2 % green tea group displayed lower body weight (p < 0.01), body fat, liver weight (p < 0.05), hepatic lipid status and higher lean mass. The coffee group displayed lower body fat (p < 0.05) and the cocoa group lower plasma glucose (p < 0.05), but this was not substantiated by other parameters. Results of study indicate the anti-obesity potential of green tea when consumed as 2 % (w/w %) of the diet probably by lipolysis and decreased systemic inflammation resulting from consumption of a high-fat, high-sucrose diet in obesity-prone strain of mice.

The antioxidant activity of cocoa

In this study we aimed to determine the antioxidant effects of Cocoa ( Theobroma cacao L.), which is commonly used in both chocolate and cocoa drinks all over the world , on lipid peroxidation, alanine transaminase (ALT), aspartate transaminase (AST), glutathione (GSH) and protein oxidation levels in carbon tetrachloride (CCl4 ) treated for male Wistar rats. Two control groups and one treatment group of rats were formed. The control groups were nourished with a standard diet, while the Cocoa group was nourished with standard diet which was enriched with % 6 by weight dried Cocoa powder. After three months, a single dose of carbon tetrachloride (CCl4 ) was performed in Control II (CCl4 ) and Cocoa groups (1ml/kg, as 20% in olive oil) intraperitoneally and single dose of olive oil was administered (1ml/kg,i.p.) in the same way as rats in Control I group. They were sacrificed two hours later. Lipid peroxide levels in liver, protein oxidation in liver, glutathione levels in liver, ALT and AST in plasma were measured. Cocoa decreased liver lipid peroxide, liver glutathione levels and plasma ALT and AST activities previously increased by CCl4 treatment, to the Control I levels The protein oxidation levels in the rats in the Cocoa group compared with the rats in CCl4 treated control group were found to have significantly lessened. These fi ndings suggest that cocoa has strong antioxidant activity because of the fact that cocoa inhibits liver injury.

A double-blind, placebo-controlled, randomized trial of the effects of dark chocolate and cocoa on variables associated with neuropsychological functioning and cardiovascular health: clinical findings from a sample of healthy, cognitively intact older adults

BackgroundIn recent years, there has been increased interest in the potential health-related benefits of antioxidant- and phytochemical-rich dark chocolate and cocoa. ObjectiveThe objective of the study was to examine the short-term (6 wk) effects of dark chocolate and cocoa on variables associated with neuropsychological functioning and cardiovascular health in healthy older adults. DesignA double-blind, placebo-controlled, fixed-dose, parallel-group clinical trial was used. Participants (n = 101) were randomly assigned to receive a 37-g dark chocolate bar and 8 ounces (237 mL) of an artificially sweetened cocoa beverage or similar placebo products each day for 6 wk. ResultsNo significant group (dark chocolate and cocoa or placebo)-by-trial (baseline, midpoint, and end-of-treatment assessments) interactions were found for the neuropsychological, hematological, or blood pressure variables examined. In contrast, the midpoint and end-of-treatment mean pulse rate assessments in the dark chocolate and cocoa group were significantly higher than those at baseline and significantly higher than the midpoint and end-of-treatment rates in the control group. Results of a follow-up questionnaire item on the treatment products that participants believed they had consumed during the trial showed that more than half of the participants in both groups correctly identified the products that they had ingested during the experiment. ConclusionsThis investigation failed to support the predicted beneficial effects of short-term dark chocolate and cocoa consumption on any of the neuropsychological or cardiovascular health-related variables included in this research. Consumption of dark chocolate and cocoa was, however, associated with significantly higher pulse rates at 3- and 6-wk treatment assessments.

A Comparative Study of Iron Bioavailability from Cocoa Supplemented with Ferric Pyrophosphate or Ferrous Fumarate in Rats

Background: Food iron fortification can be a good strategy to prevent iron deficiency. Iron bioavailability from cocoa powder enriched with ferric pyrophosphate encapsulated in liposomes or ferrous fumarate was assessed in rats. Methods: Three groups of rats consumed during 28 days either a control diet or two diets prepared with ferric pyrophosphate- or ferrous fumarate-enriched cocoa powder as the unique source of iron. Body weight and food intake were monitored and last-week feces were collected. On day 28, animals were sacrificed and livers and spleens were removed. Hemoglobin and total iron binding capacity (TIBC) were determined. Results: There were no significant differences in body weight and food intake. Apparent iron absorption and % absorption/intake were significantly lower in rats consuming enriched cocoa compared to the control group, without significant differences due to the iron form. Enriched cocoa groups showed significantly lower spleen iron content and concentration than the control. Liver iron was lower in the ferric pyrophosphate group compared to the other two groups. Hemoglobin and TIBC values showed a deficient iron status in ferric pyrophosphate rats. Conclusion: Cocoa powder is a good vehicle for iron fortification when enriched with ferrous fumarate compared to ferric pyrophosphate encapsulated in liposomes.

Continuous intake of polyphenolic compounds containing cocoa powder reduces LDL oxidative susceptibility and has beneficial effects on plasma HDL-cholesterol concentrations in humans

Background:Cocoa powder is rich in polyphenols such as catechins and procyanidins and has been shown in various models to inhibit LDL oxidation and atherogenesis.Objective:We examined whether long-term intake of cocoa powder alters plasma lipid profiles in normocholesterolemic and mildly hypercholesterolemic human subjects.Design:Twenty-five subjects were randomly assigned to ingest either 12 g sugar/d (control group) or 26 g cocoa powder and 12 g sugar/d (cocoa group) for 12 wk. Blood samples were collected before the study and 12 wk after intake of the test drinks. Plasma lipids, LDL oxidative susceptibility, and urinary oxidative stress markers were measured.Results:At 12 wk, we measured a 9% prolongation from baseline levels in the lag time of LDL oxidation in the cocoa group. This prolongation in the cocoa group was significantly greater than the reduction measured in the control group (−13%). A significantly greater increase in plasma HDL cholesterol (24%) was observed in the cocoa group than in the control group (5%). A negative correlation was observed between plasma concentrations of HDL cholesterol and oxidized LDL. At 12 wk, there was a 24% reduction in dityrosine from baseline concentrations in the cocoa group. This reduction in the cocoa group was significantly greater than the reduction in the control group (−1%).Conclusion:It is possible that increases in HDL-cholesterol concentrations may contribute to the suppression of LDL oxidation and that polyphenolic substances derived from cocoa powder may contribute to an elevation in HDL cholesterol.

Long-Term Ingestion of High Flavanol Cocoa Provides Photoprotection against UV-Induced Erythema and Improves Skin Condition in Women

Dietary antioxidants contribute to endogenous photoprotection and are important for the maintenance of skin health. In the present study, 2 groups of women consumed either a high flavanol (326 mg/d) or low flavanol (27 mg/d) cocoa powder dissolved in 100 mL water for 12 wk. Epicatechin (61 mg/d) and catechin (20 mg/d) were the major flavanol monomers in the high flavanol drink, whereas the low flavanol drink contained 6.6 mg epicatechin and 1.6 mg catechin as the daily dose. Photoprotection and indicators of skin condition were assayed before and during the intervention. Following exposure of selected skin areas to 1.25 x minimal erythemal dose (MED) of radiation from a solar simulator, UV-induced erythema was significantly decreased in the high flavanol group, by 15 and 25%, after 6 and 12 wk of treatment, respectively, whereas no change occurred in the low flavanol group. The ingestion of high flavanol cocoa led to increases in blood flow of cutaneous and subcutaneous tissues, and to increases in skin density and skin hydration. Skin thickness was elevated from 1.11 +/- 0.11 mm at wk 0 to 1.24 +/- 0.13 mm at wk 12; transepidermal water loss was diminished from 8.7 +/- 3.7 to 6.3 +/- 2.2 g/(h x m2) within the same time frame. Neither of these variables was affected in the low flavanol cocoa group. Evaluation of the skin surface showed a significant decrease of skin roughness and scaling in the high flavanol cocoa group compared with those at wk 12. Dietary flavanols from cocoa contribute to endogenous photoprotection, improve dermal blood circulation, and affect cosmetically relevant skin surface and hydration variables.

Chronic Consumption of Flavanol-rich Cocoa Improves Endothelial Function and Decreases Vascular Cell Adhesion Molecule in Hypercholesterolemic Postmenopausal Women

Endothelial dysfunction characterizes many disease states including subclinical atherosclerosis. The consumption of flavanol-rich cocoa and cocoa-based products has been shown to improve endothelial function in both compromised and otherwise normal, healthy individuals when administered either acutely or over a period of several days, or weeks. Women experience increased risk for cardiovascular disease after menopause, which can be associated with endothelial dysfunction. Whether a flavanol-rich cocoa-based product can improve endothelial function in hypercholesterolemic postmenopausal women is not known. The purpose of the present study was to determine whether chronic dietary administration of flavanol-rich cocoa improves endothelial function and markers of cardiovascular health in hypercholesterolemic postmenopausal women. Thirty-two postmenopausal hypercholesterolemic women were randomly assigned to consume a high-flavanol cocoa beverage (high cocoa flavanols (CF)--446 mg of total flavanols), or a low-flavanol cocoa beverage (low CF--43 mg of total flavanols) for 6 weeks in a double-blind study (n=16 per group). Endothelial function was determined by brachial artery-reactive hyperemia. Plasma was analyzed for lipids (total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol), hormones (follicle-stimulating hormone), total nitrate/nitrite, activation of cellular adhesion markers (vascular cell adhesion molecule 1, intercellular adhesion molecule 1, E-Selectin, P-Selectin), and platelet function and reactivity. Changes in these plasma markers were then correlated to brachial reactivity. Brachial artery hyperemic blood flow increased significantly by 76% (P<0.05 vs. baseline) after the 6-week cocoa intervention in the high CF group, compared with 32% in the low CF cocoa group (P=ns vs. baseline). The 2.4-fold increase in hyperemic blood flow with high CF cocoa closely correlated (r2=0.8) with a significant decrease (11%) in plasma levels of soluble vascular cell adhesion molecule-1. Similar responses were not observed after chronic use of low CF. There were no significant differences between high and low CF in other biochemical markers and parameters measured. This study is the first to identify beneficial vascular effects of flavanol-rich cocoa consumption in hypercholesterolemic postmenopausal women. In addition, our results suggest that reductions in plasma soluble vascular cell adhesion molecule-1 after chronic consumption of a flavanol-rich cocoa may be mechanistically linked to improved vascular reactivity.

Daily cocoa intake reduces the susceptibility of low-density lipoprotein to oxidation as demonstrated in healthy human volunteers

Nine male volunteers were given 36 g of cocoa powder (containing 2610 mg of polyphenols) per day with sugar and 6 volunteers received an equivalent amount of sugar for 2 weeks. Conjugated diene production in LDL induced by 2-2′ azobis(4-methoxy-2,4-dimethyl-valeronitrile) (V-70) and copper ion were evaluated. The lag time was significantly prolonged at 1 and 2 weeks in V-70 and at 2 weeks in copper ion after cocoa powder consumption. The level of excretion of epicatechin in urine was significantly higher in the cocoa group than that in the control group. In conclusion, the antioxidants in cocoa powder might be absorbed and increase the resistance of human LDL to oxidation.