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Cocoa flavan-3-ols affect endothelium-dependent responses in resistance vessels and microcirculation has received little attention. We tested the effects of dark chocolate consumption (396 mg total flavanols/day for 3 days) in two Groups of 10 men (18–25 years; non-smokers) each comprising equal numbers of White European (WE) and South Asian (SA) ethnicity. In Group 1, dark chocolate did not affect reactive hyperaemia in forearm muscle, but augmented muscle dilatation evoked by acute mental stress, and reactive hyperaemia and acetylcholine (ACh)-evoked dilatation in cutaneous microcirculation. Conversely, in Group 2, chocolate did not affect cutaneous reactive hyperaemia or ACh-evoked dilatation, but these responses were blunted in Group 1 relative to Group 2. Further, when Groups 1 and 2 were combined, responses were blunted in SAs relative to WEs, augmented by chocolate in SAs only. In Group 2 individuals whose ACh-evoked dilatation was attenuated by nitric oxide synthase (NOS) inhibition, ACh-evoked dilatation was not altered after chocolate, but the attenuating effect of NOS inhibition was lost. Conversely, in Group 2 individuals whose ACh-evoked dilatation was enhanced by NOS inhibition, ACh-evoked dilatation was also augmented by chocolate. We propose that in resistance and microvessels of young men, cocoa flavan-3-ols preferentially augment endothelium-dependent dilator responses whose responses are depressed by familial and lifestyle factors more prevalent in SAs than Wes. Flavan-3-ols may facilitate the NOS pathway but also influence other endothelium-dependent dilators. [ABSTRACT FROM AUTHOR]

Studies indicated that cocoa-based products effectively mitigate the risks associated with metabolic syndrome (MetS), however, the effect varies based on cocoa types, dosages, and study durations. This review aimed to determine the flavanol-rich cocoa consumption on MetS outcomes within the last decade (2013-2023), adhering to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Seven randomized-controlled trials (RCTs) used cocoa-based products containing 0.3-1680 mg flavanol monomers and 3.5-1270 mg procyanidins. Cocoa-based products beneficially reduced glycemic response, blood pressure and lipid profiles. However, this review highlights little evidence pinpointing the best cocoa products type and required dosage for the observed effects. Further intervention aiming to improve MetS should justify the selection and concentration of flavanols (monomers and procyanidins). A robust study design should consider registering the trials before study commencement, consider multicenter RCT trials, and adjust for potential covariates that might "masked" the outcomes. [ABSTRACT FROM AUTHOR]

Hyperactivation of blood platelets, one of the causes of heart attack, and other cardiovascular diseases (CVDs), is influenced by various dietary components, including phenolic compounds from vegetables, fruits, teas, wines, cocoa and its products, including chocolate. The present paper sheds new light on the effect of cocoa and its products, especially dark chocolate, on the number and function of blood platelets, and the anti-platelet activity of their constituent phenolic compounds. A review was performed of papers identified in various electronic databases, including PubMed, Science Direct, Scopus, Web of Knowledge, and Google Scholar, with the aim of determining whether their anti-platelet activity may serve as part of a sweet strategy in countering CVDs. Various studies demonstrate that cocoa consumption, especially in the form of dark chocolate, with a high flavanol concentration, has anti-platelet activity and may play a significant role in cardioprotection; they also note that cocoa consumption may be a good strategy in diminishing cardiovascular risk, including hyperactivation of blood platelets. [ABSTRACT FROM AUTHOR]

Hyperactivation of blood platelets, one of the causes of heart attack, and other cardiovascular diseases (CVDs), is influenced by various dietary components, including phenolic compounds from vegetables, fruits, teas, wines, cocoa and its products, including chocolate. The present paper sheds new light on the effect of cocoa and its products, especially dark chocolate, on the number and function of blood platelets, and the anti-platelet activity of their constituent phenolic compounds. A review was performed of papers identified in various electronic databases, including PubMed, Science Direct, Scopus, Web of Knowledge, and Google Scholar, with the aim of determining whether their anti-platelet activity may serve as part of a sweet strategy in countering CVDs. Various studies demonstrate that cocoa consumption, especially in the form of dark chocolate, with a high flavanol concentration, has anti-platelet activity and may play a significant role in cardioprotection; they also note that cocoa consumption may be a good strategy in diminishing cardiovascular risk, including hyperactivation of blood platelets.

The objective of this study was to optimize different vacuum drying conditions for cocoa pod husk and cocoa bean shell in order to enhance these by-products for commercial applications. To carry out the optimization, the response surface methodology was applied using a Box–Behnken experimental design with 15 experiments for which different conditions of temperature (X1), drying time (X2) and vacuum pressure (X3) were established. The response variables were the content of total polyphenols, the content of flavanols and the radical scavenging activity evaluated in the extracts of the different experiments. Temperature (50–70 °C), drying time (3–12 h) and vacuum pressure (50–150 mbar) were considered as independent variables. The main factors affecting the response variables were temperature, followed by vacuum pressure. For the content of polyphenols, the optimal response values predicted for the cocoa pod husk was 11.17 mg GAE/g with a confidence limit (95%) of 9.05 to 13.28 mg GAE/g (optimal conditions: 65 °C, 8 h and 75 mbar), while for the cocoa bean shell cocoa was 29.61 mg GAE/g with a confidence limit (95%) of 26.95 to 32.26 mg GAE/g (optimal conditions: 50 °C, 5 h and 100 mbar). Therefore, results of this study suggest a high content of phenolic compounds obtained from these by-products that show relevance as functional ingredients for application in the food, nutraceutical, and cosmeceutical industries. [ABSTRACT FROM AUTHOR]

(1) Background: Cocoa's healthy benefits may be attributed to the potent antioxidant activity of cocoa polyphenols, mainly flavanols, which have been characterised as existing in a high concentration in cocoa. However, the phenolic composition of cocoa and cocoa-derived products is highly variable, and manufacturing processes might significantly reduce their phenolic content. For that reason, the full characterisation of cocoa and cocoa-derived products before evaluating their bioactivity is crucial. The aim of this review is to analyse the available evidence on the effect of flavanol-fortified cocoa-derived products on human health. (2) Methods: Forty-eight clinical trials focused on the health effect of consuming flavanol-fortified drinks, bars and chocolate have been reviewed, with a total of 1523 subjects. (3) Results: Although studies differ widely in methodology, dosage, duration, and target population, beneficial effects of flavanol-rich cocoa consumption have been observed at doses ranging from 45.3 mg/d to 1078 mg/d, especially on cardiovascular health and cognitive function. (4) Conclusions: Considering the high consumption and acceptability of cocoa and cocoa-derived products, the fortification of cocoa products as well as other highly consumed foods with cocoa flavanols could be an effective strategy for health promotion. [ABSTRACT FROM AUTHOR]

Background: Cocoa and dark chocolate contain polyphenols and several other positive components, and diets containing balanced amounts of these substances have beneficial effects on human health. In this review article provided a comprehensive and fruitful explanations about cocoa and chocolate as a world popular food, their positive and constructive properties in the physiological processes of the body, especially diabetes. Materials and Methods: In this study has been used the method of library collection, search in various texts and valid scientific articles. Results: Overall, balanced consumption of appropriate doses of cocoa and chocolate, especially dark chocolate, in addition to slowing the progression of diabetes and its preventive effects, contains many other positive effects, include increasing the potential to prevent cardiovascular disease and increasing the efficiency and capacity of the heart and kidneys, increasing endorphins and delivering it to the brain and increasing the body's sense of pleasure and energy, improving digestion. As a result, they help improve gastrointestinal function, increase good cholesterol (HDL) and decrease bad cholesterol (LDL), and have anti-inflammatory and anti-cancer properties. Conclusion: Consumption of chocolate, especially dark type, if it is appropriate, due to its beneficial components such as flavanols, can have positive effects on many body processes, especially blood sugar control, and prevent diabetes.

The aim of this study was to establish the influence of flavonoid-enriched diets on the immune alterations induced by an intensive training and a final exhaustion test in rats. A flavanol-enriched diet (with 10% cocoa, C10 diet) and a flavanol and flavanone-enriched diet (C10 plus 0.5% hesperidin, CH diet) were used. Lewis rats were fed either a standard diet, C10 diet or CH diet while they were submitted to an intensive running training on a treadmill. After 6 weeks, samples were obtained 24 h after performing a regular training (T groups) and after carrying out a final exhaustion test (TE groups). The C10 diet attenuated the increase in plasma cortisol induced by exhaustion, while both the C10 and the CH diets prevented the alterations in the spleen Th cell proportion. The experimental diets also induced an increase in serum immunoglobulin concentration and an enhancement of spleen natural killer cytotoxicity, which may be beneficial in situations with a weakened immunity. Most of the effects observed in the CH groups seem to be due to the cocoa content. Overall, a dietary intervention with flavonoids enhances immune function, partially attenuating the alterations in systemic immunity induced by intensive training or exhausting exercise. [ABSTRACT FROM AUTHOR]

Despite the health benefits associated with the ingestion of the bioactive compounds in cocoa, the high concentrations of polyphenols and methylxanthines in the raw cocoa beans negatively influence the taste, confer the astringency and bitterness, and affect the stability and digestibility of the cocoa products. It is, therefore, necessary to process cocoa beans to develop the characteristic color, taste, and flavor, and reduce the astringency and bitterness, which are desirable in cocoa products. Processing, however, affects the composition and quantities of the bioactive compounds, resulting in the modification of the health-promoting properties of cocoa beans and chocolate. In this advanced review, we sought to better understand the effect of cocoa's transformational process into chocolate on polyphenols and methylxanthine and the mechanism of action of the original flavanols and methylxanthines. More data on the cocoa processing effect on cocoa bioactives are still needed for better understanding the effect of each processing step on the final polyphenolic and methylxanthine composition of chocolate and other cocoa products. Regarding the mechanisms of action, theobromine acts through the modulation of the fatty acid metabolism, mitochondrial function, and energy metabolism pathways, while flavanols mainly act though the protein kinases and antioxidant pathways. Both flavanols and theobromine seem to be involved in the nitric oxide and neurotrophin regulation. [ABSTRACT FROM AUTHOR]

Chocolate is the most sought-after delicacy worldwide. Cocoa has been incorporated in various forms, into both foods and cosmetics. Cocoa has not only been used as gourmet food but has also served as medicine for ages. Chocolates are always in vogue and remain a testament to the cocoa bean, but the benefits attained from commercial chocolates remain a fallacy. On a positive note, chocolate has its health benefits but its effects on the skin need a better understanding. Hence this article focuses on the dermatological effects of cocoa, and its use as a dermatological therapeutic agent. [ABSTRACT FROM AUTHOR]

(1) Background: Cocoa’s healthy benefits may be attributed to the potent antioxidant activity of cocoa polyphenols, mainly flavanols, which have been characterised as existing in a high concentration in cocoa. However, the phenolic composition of cocoa and cocoa-derived products is highly variable, and manufacturing processes might significantly reduce their phenolic content. For that reason, the full characterisation of cocoa and cocoa-derived products before evaluating their bioactivity is crucial. The aim of this review is to analyse the available evidence on the effect of flavanol-fortified cocoa-derived products on human health. (2) Methods: Forty-eight clinical trials focused on the health effect of consuming flavanol-fortified drinks, bars and chocolate have been reviewed, with a total of 1523 subjects. (3) Results: Although studies differ widely in methodology, dosage, duration, and target population, beneficial effects of flavanol-rich cocoa consumption have been observed at doses ranging from 45.3 mg/d to 1078 mg/d, especially on cardiovascular health and cognitive function. (4) Conclusions: Considering the high consumption and acceptability of cocoa and cocoa-derived products, the fortification of cocoa products as well as other highly consumed foods with cocoa flavanols could be an effective strategy for health promotion.

Cocoa and coffee are natural sources of phenolic compounds, which are degraded during beans processing of both. For this reason, there is an interest in obtaining extracts of these bio compounds. The aim of this study was uncovering the radical scavenging activity (AC) of mixtures of cocoa and coffee extracts, and the bioavailability of their phenolic compounds, resorting to in vitro models: DPPH and ORAC antioxidant methods, and the characterization of the trans-epithelial transport of cocoa and coffee phenolics through Caco-2 cells monolayer model of the intestinal barrier. The cocoa displayed a higher AC than the coffee regarding both DPPH and ORAC assays, while the mixtures increased in parallel with the percentage of cocoa in the blends. The combination index was calculated to set up the type of interaction in the cocoa-coffee mixtures, obtaining that the mixture 25:75 was moderately antagonistic, 50:50 nearly additive, and 75:25 slightly synergistic. The absorption efficiency of the cocoa phenolic compounds was between 87.9%–97.4%, in the coffee compounds was 100%. The mixtures varied according to the proportion of cocoa and coffee. The results obtained allowed concluding that the phenolic compounds present in cocoa and coffee, respectively, are featured by high bioavailability and a valuable antioxidant capacity, while no pattern was found in the mixtures concerning the real benefit of using them combined. [ABSTRACT FROM AUTHOR]

Background: Cocoa flavanols and procyanidins (CF) are flavonoids whose consumption is associated with health benefits, resulting in increasing attention from consumers, industry, researchers, and regulators. Methods that can provide appropriate characterization and quantification of the distinct mixture found in cocoa-based products thus offer important scientific and commercial value. Objective: This study validated the precision of AOAC Official Method of AnalysisSM 2020.05, which measures CF with a degree of polymerization DP1-7. Method: Method precision (repeatability and reproducibility) was evaluated for seven cocoa matrixes in blind duplicates with total CF content from 1.0 to 500 mg/g. Ten of the 12 laboratories from multiple sectors invited to implement the method returned data for statistical analysis. Precision was evaluated per AOAC INTERNATIONAL guidelines for collaborative studies using RSDr and RSDR as indicators of method repeatability and reproducibility. Results: RSDr ranged from 1.6 to 4.8%, and RSDR ranged from 5.8 to 22.4%, demonstrating excellent within-laboratory repeatability and good method precision across different laboratories. RSDR values were below 10% with the exception of chocolate, potentially due to very low CF content and sampling inhomogeneity. Conclusions: These data demonstrate that acceptable method repeatability and reproducibility is achieved when measuring cocoa flavanols and procyanidins using AOAC Method 2020.05 and support the advancement of the AOAC Official Method of Analysis status to Final Action for evaluated matrixes. Highlights: This collaborative study evaluated the repeatability and reproducibility of AOAC Official Method of Analysis 2020.05. [ABSTRACT FROM AUTHOR]

Cognitive decline is a common problem in older individuals, often exacerbated by neurocognitive conditions, such as vascular dementia and Alzheimer's disease, which heavily affect people's lives and exert a substantial toll on healthcare systems. Currently, no cure is available, and commonly used treatments are aimed at limiting the progressive loss of cognitive functions. The absence of effective pharmacological treatments for the cognitive decline has led to the search for lifestyle interventions, such as diet and the use of nutraceuticals that can prevent and limit the loss of cognition. Cocoa and chocolate are foods derived from cocoa beans, commonly used in the population and with good acceptability. The purpose of this review was to collect current experimental evidence regarding the neuroprotective effect of chocolate and cocoa (or derived molecules) in the elderly. From a systematic review of the literature, 9 observational studies and 10 interventional studies were selected, suggesting that the biomolecules contained in cocoa may offer promising tools for managing cognitive decline, if provided in adequate dosages and duration of treatment. However, the molecular mechanisms of cocoa action on the central nervous system are not completely understood. [ABSTRACT FROM AUTHOR]

Introduction: Cocoa and chocolate products have recently been recognized as a rich source of flavonoids, mainly flavanols, powerful antioxidants and anti-inflammatory agents that enter and accumulate in the brain regions involved in learning and memory, especially the hippocampus. The neurobiological activities of flavanols are thought to occur in two main ways: (i) through direct interaction with cellular cascades inducing the expression of neuroprotective and neuroregulatory proteins that promote neurogenesis, neuronal function, and brain connectivity, and (ii) through enhancement of blood flow and angiogenesis in the brain and sensory systems. Therefore, the aim of this study was to summarize the results on the impact of cocoa consumption on cognitive function and brain health. Method: The present article provides relevant articles available on the mechanism of action of cocoa and its bioactive ingredients on the brain. These studies were discovered via searching PubMed, Web of science, and Embase databases from 2010 to 2023. After removing duplicate articles, fifteen articles met the inclusion/exclusion criteria of our review and 6 of them were included. Result: The epicatechin content in cocoa is primarily responsible for its favorable effects on vascular endothelium by influencing the regulation of nitric oxide production, which increases cerebral blood flow to improve cognitive function and attention. In addition, improved cognitive function has been reported to be associated with increased levels of blood brain-derived neurotrophic factor (BDNF), a protein associated with neuronal growth levels. In the brain, BDNF stimulates synaptic plasticity and neurogenesis, and plays an important role in brain regions involved in learning and memory, such as the hippocampus and the sub ventricular zone. Conclusion: These results support the idea that daily servings of dark chocolate (35 g) may provide benefits to the brain in healthy consumers. There is clear evidence that cocoa flavonoids can significantly improve cognitive function in humans, possibly through mechanisms such as improving cerebral blood flow. [ABSTRACT FROM AUTHOR]

The aim of this study was to establish the influence of flavonoid-enriched diets on the immune alterations induced by an intensive training and a final exhaustion test in rats. A flavanol-enriched diet (with 10% cocoa, C10 diet) and a flavanol and flavanone-enriched diet (C10 plus 0.5% hesperidin, CH diet) were used. Lewis rats were fed either a standard diet, C10 diet or CH diet while they were submitted to an intensive running training on a treadmill. After 6 weeks, samples were obtained 24 h after performing a regular training (T groups) and after carrying out a final exhaustion test (TE groups). The C10 diet attenuated the increase in plasma cortisol induced by exhaustion, while both the C10 and the CH diets prevented the alterations in the spleen Th cell proportion. The experimental diets also induced an increase in serum immunoglobulin concentration and an enhancement of spleen natural killer cytotoxicity, which may be beneficial in situations with a weakened immunity. Most of the effects observed in the CH groups seem to be due to the cocoa content. Overall, a dietary intervention with flavonoids enhances immune function, partially attenuating the alterations in systemic immunity induced by intensive training or exhausting exercise.

Flavanols, a subgroup of polyphenols, are secondary metabolites with antioxidant properties naturally produced in various plants (e.g., green tea, cocoa, grapes, and apples); they are a major polyphenol class in human foods and beverages, and have recognized effect on maintaining human health. Therefore, it is necessary to evaluate their changes (i.e., oxidation, polymerization, degradation, and epimerization) during various physical processing (i.e., heating, drying, mechanical shearing, high‐pressure, ultrasound, and radiation) to improve the nutritional value of food products. However, the roles of flavanols, in particular for their polymerized forms, are often underestimated, for a large part because of analytical challenges: they are difficult to extract quantitatively, and their quantification demands chemical reactions. This review examines the existing data on the effects of different physical processing techniques on the content of flavanols and highlights the changes in epimerization and degree of polymerization, as well as some of the latest acidolysis methods for proanthocyanidin characterization and quantification. More and more evidence show that physical processing can affect content but also modify the structure of flavanols by promoting a series of internal reactions. The most important reactivity of flavanols in processing includes oxidative coupling and rearrangements, chain cleavage, structural rearrangements (e.g., polymerization, degradation, and epimerization), and addition to other macromolecules, that is, proteins and polysaccharides. Some acidolysis methods for the analysis of polymeric proanthocyanidins have been updated, which has contributed to complete analysis of proanthocyanidin structures in particular regarding their proportion of A‐type proanthocyanidins and their degree of polymerization in various plants. However, future research is also needed to better extract and characterize high‐polymer proanthocyanidins, whether in their native or modified forms. [ABSTRACT FROM AUTHOR]

Exercise-induced muscle damage (EIMD) is associated with oxidative stress and inflammation, muscle soreness, and reductions in muscle function. Cocoa flavanols (CF) are (poly)phenols with antioxidant and anti-inflammatory effects and thus may attenuate symptoms of EIMD. The purpose of this narrative review was to collate and evaluate the current literature investigating the effect of CF supplementation on markers of exercise-induced oxidative stress and inflammation, as well as changes in muscle function, perceived soreness, and exercise performance. Acute and sub-chronic intake of CF reduces oxidative stress resulting from exercise. Evidence for the effect of CF on exercise-induced inflammation is lacking and the impact on muscle function, perceived soreness and exercise performance is inconsistent across studies. Supplementation of CF may reduce exercise-induced oxidative stress, with potential for delaying fatigue, but more evidence is required for any definitive conclusions on the impact of CF on markers of EIMD. [ABSTRACT FROM AUTHOR]

Cocoa flavanols and procyanidins (CFs), natural dietary bioactives, have been studied extensively over the past two decades for their potential health benefits. Research on their safety and efficacy is critically dependent upon on the ability to reliably characeterize the research materials that are utilized, and with growing consumer availability of CF-based products, reliable methods for the detection of potential adulteration are of increasing importance. This research focused on the development of a high performance liquid chromatography-tandem mass spectrometry method (HPLC-MS2) using primary standards and 13C-labelled procyanidins as internal standards. The ability of MS2 detection to discriminate A- and B-type procyanidins was demonstrated. Method performances were validated for degrees of polymerization up to four in seven model food matrices. Accuracy ranged from 90.9 to 125.4% and precision was < 10% at lower concentrations. Finally, the method was applied to cocoa-based samples and compared to the AOAC 2020.05 analytical protocol, supporting the use of NIST 8403 as reference material for HPLC-MS2 analysis. [ABSTRACT FROM AUTHOR]

Cognitive decline is a common problem in older individuals, often exacerbated by neurocognitive conditions, such as vascular dementia and Alzheimer’s disease, which heavily affect people’s lives and exert a substantial toll on healthcare systems. Currently, no cure is available, and commonly used treatments are aimed at limiting the progressive loss of cognitive functions. The absence of effective pharmacological treatments for the cognitive decline has led to the search for lifestyle interventions, such as diet and the use of nutraceuticals that can prevent and limit the loss of cognition. Cocoa and chocolate are foods derived from cocoa beans, commonly used in the population and with good acceptability. The purpose of this review was to collect current experimental evidence regarding the neuroprotective effect of chocolate and cocoa (or derived molecules) in the elderly. From a systematic review of the literature, 9 observational studies and 10 interventional studies were selected, suggesting that the biomolecules contained in cocoa may offer promising tools for managing cognitive decline, if provided in adequate dosages and duration of treatment. However, the molecular mechanisms of cocoa action on the central nervous system are not completely understood.

In healthy people, cocoa-derived flavanols (CF) improve blood pressure (BP). This meta-analysis investigates whether CF also affect BP in diabetic patients. PubMed, Web of Science, and Embase were consulted to retrieve eligible randomized controlled trials. A random-effects model and the Grades of Recommendation, Assessment, Development and Evaluation (GRADE)-approach were used for analyses and quality of evidence respectively. Of 267 citations, 11 trials were identified, studying either type 2 diabetic populations only (subgroup A) or type 2 diabetic patients plus non-diabetic subjects with increased cardiovascular risk (subgroup B1) or type 1 plus type 2 diabetic patients (subgroup B2). Mid/long-term CF consumption decreased BP slightly, however, only reaching statistical significance for diastolic BP in subgroup B1 (-1.89 mmHg, 95% CI: −3.24, −0.54, I2 = 55%). Considerable heterogeneity between studies and low quality of evidence caused poor quality evidence of minimal effects of CF ingestion on BP in diabetic patients.

This article discusses the potential benefits of flavanols, a group of plant-based compounds, in reducing age-related memory loss (ARML) and cognitive decline in older adults. Flavanols, which are found in foods such as fruits, vegetables, tea, cocoa, and red wine, have antioxidant and anti-inflammatory properties that may protect against neurocognitive decline. Studies have shown that flavanols can improve memory scores, particularly in older adults with poor flavanol intake at baseline. However, there is heterogeneity in previous studies, and more evidence from large-scale longitudinal studies is needed to establish the effectiveness of flavanol-rich diets in maintaining cognitive health during aging. Despite these limitations, flavanols offer a promising avenue for supporting cognitive health and reducing the risk of neurodegenerative diseases in the elderly population. [Extracted from the article]

Background: Flavanols and procyanidins are complex bioactives found in many foods such as cocoa. As their consumption is associated with health benefits, cocoa flavanols and procyanidins are receiving increasing attention from consumers, industry, researchers, and regulators. Objective: The objective of this study is to validate a method using hydrophilic interaction chromatography (HILIC) with fluorescence detection (FLD) and a commercially available reference material for the determination of flavanols and procyanidins (CF) in cocoa-based products. Methods: Method performances were evaluated for cocoa matrices with CF content that ranged from 0.8 to 500 mg/g, which included low CF matrices (milk and dark chocolate, cocoa powder, and liquor) and high CF matrices (cocoa extract and dietary supplement products). The method was validated in a single-laboratory by determining sensitivity, selectivity, linearity, stability, robustness, accuracy, and precision for each of the matrices. Results: The method was validated for cocoa matrices with CF content that ranged from 0.8 to 500 mg/g. Accuracy ranged from 86 to 99% and repeatability (RSDr) from 1.5 to 8.6% for CF. Conclusions: Analytical performances acquired through this single-laboratory validation study for a wide range of cocoa-based matrices demonstrate that this method is fit-for-purpose for the determination of flavanols and procyanidins in cocoa-based products. Highlights: Hydrophilic interaction chromatography (HILIC) with fluorescence detection was successfully used to determine total CF content in multiple product types. Single-laboratory method validation results demonstrate that the method is fit for purpose for cocoa-based matrices containing <0.8 to 500 mg/g of CF. [ABSTRACT FROM AUTHOR]

Background: In many cocoa intervention studies, health outcomes are related to cocoa components without taking into account the bioavailability of the main bioactive components: phenolic compounds and methylxanthines. Methods: The present work associates the results of bioavailability and randomised controlled crossover studies in humans carried out with similar cocoa products, so that the main phenol and methylxanthine metabolites observed in plasma and urine are associated to the health effects observed in the chronic studies. We outstand that doses of cocoa and consumption rate used are realistic. In the bioavailability study, a conventional (CC) and a methylxanthine-polyphenol rich (MPC) cocoa product were used, whereas in the chronic study a dietary fibre-rich (DFC) and a polyphenol-rich (PC) product were studied in healthy and cardiovascular risk subjects. Results and Discussion: The main phenolic metabolites formed after CC and MPC intake, 5-(4'-hydroxyphenyl)-valerolactone-3'-sulfate, 3'-methyl-epicatechin- 5-sulfate, 4-hydroxy-5-(4'-hydroxyphenyl)valeric acid-sulfate, 5-phenyl- valerolactone-sulfate and 5-(4'-hydroxyphenyl)-valerolactone-3'-glucuronide, may contribute to the changes in cholesterol (and indirectly HDL-cholesterol) observed after the regular intake of both DFC and PC, in healthy and cardiovascular risk subjects, whereas 7-methylxanthine (the main cocoa methylxanthine metabolite) and theobromine, together with its content in insoluble dietary fibre, may be responsible for the decrease of IL-1β and hypoglycemic effects observed with DFC. With both phenolic and methylxanthine metabolites a strong dose-response effect was observed. Conclusion: After the regular consumption of both DFC and PC, positive changes were observed in volunteer's lipid profile, which may be related to the long-lasting presence of colonic phenolic metabolites in blood. In contrast, the anti-inflammatory and hypoglycemic effects were only observed with DFC, and these may be related to methylxanthine metabolites, and it is likely that insoluble dietary fibre may have also played a role. [ABSTRACT FROM AUTHOR]

Oxidative stress may cause functional disorders of vascular endothelia which can lead to endothelial apoptosis and thus alter the function and structure of the vascular tissues. Plant antioxidants protect the endothelium against oxidative stress and then become an effective option to treat vascular diseases. Cocoa flavanols have been proven to protect against oxidative stress in cell culture and animal models. In addition, epidemiological and interventional studies strongly suggest that cocoa consumption has numerous beneficial effects on cardiovascular health. The objective of this study was to test the chemo-protective effect of realistic concentrations of a cocoa phenolic extract and its main monomeric flavanol epicatechin on cultured human endothelial cells submitted to an oxidative challenge. Both products efficiently restrained stress-induced reactive oxygen species and biomarkers of oxidative stress such as carbonyl groups and malondialdehyde, and recovered depleted glutathione, antioxidant defences and cell viability. Our results demonstrate for the first time that a polyphenolic extract from cocoa and its main flavonoid protect human endothelial cells against an oxidative insult by modulating oxygen radical generation and antioxidant enzyme and non-enzyme defences. [ABSTRACT FROM AUTHOR]

The effects of cocoa-derived polyphenols on cognitive functions have been analyzed through numerous studies using different interventions (doses, vehicles, time frame, cognition tests, and characteristics of participants) which may hamper the interpretation and comparison of findings across investigations. Thus, a systematic review was conducted to analyze the effects of cocoa-derived polyphenols intake on human cognition and discuss the methodological aspects that may contribute to the heterogeneity of findings. Randomized clinical trials evaluating the effect of cocoa polyphenols on cognitive function in healthy subjects were selected according to selection criteria. Twelve studies were selected. Quality was assessed according to the Cochrane risk for bias tool. The most common risk for bias was the lack of information about the sequence generation process. Effects on cognitive function were observed after consumption of 50 mg/day of (-)-epicatechin and in studies using a component-matched placebo and cocoa as the polyphenol vehicle given to healthy adults (18-50 years). Memory (n = 5) and executive function (n = 4) showed the most significant effects with medium and large effect sizes after intake of intermediate doses of cocoa flavanols (500-750 mg/day). Overall, this set of studies suggest a positive effect of cocoa polyphenols on memory and executive function. However, the available evidence is very diverse and future studies may address the identified sources of variation to strengthen current evidence on this promising field. [ABSTRACT FROM AUTHOR]

Cocoa contains a range of chemicals that can interact with cells and tissue components, providing protection against the development and improvement of pathological conditions. The most important effects of cocoa and chocolate are related to cardiovascular disease. Due to the high content of flavonoids, it has numerous positive effects on cardiovascular diseases, including: antioxidant, anti-inflammatory, antithrombotic effects, and possibly increased HDL, lowering blood pressure and improving endothelial function. The beneficial effects of cocoa are most likely a consequence of reducing oxidative stress and increasing the bioavailability of nitric oxide. Although many positive effects of chocolate on the cardiovascular system have been proven, precautions in its use are mandatory. [ABSTRACT FROM AUTHOR]

Abstract Background Cocoa-containing food products are getting recognized as a blood pressure (BP) controlling food choice. The beneficial effect of cocoa on BP is attributable to its high content of flavanols, which are composed of epicatechin, catechin, and procyanidins. The main mechanism by which cocoa flavanols reduce BP is increasing nitric oxide. With a high prevalence of hypertension in the North America, a health claim petition on cocoa flavanols may be considered within the US framework, in order to allow people to make better food choices. Scope and approach The objective of this review is to evaluate whether there is sufficient evidence to support a BP-lowering health claim for cocoa flavanols under the US Food and Drug Administration (FDA)'s jurisdiction. Seventeen human intervention studies examining the effect of cocoa flavanols on BP were identified, with nine studies in normotensive participants, and eight studies in hypertensive participants. All studies were assessed for methodological quality based on the outline provided by the FDA. Key findings and conclusions Nine out of the 17 studies reported a BP-lowering effect of cocoa flavanols. This indicates the evidence of cocoa flavanols on BP reduction was conflicting. Only a few studies presented as high quality. The optimal dose of cocoa flavanols and long-term effect of cocoa flavanols on BP remain to be clarified. Therefore, within the US health claims system, an authorized health claim should not be passed, but a qualified health claim that is supported by low evidence may be attributed to cocoa flavanols and BP reduction. Highlights • Consumption of cocoa flavanols has been shown to reduce BP. • The relationship between cocoa flavanols and BP is examined based on the US FDA health claims systems. • Inconsistent evidence exists on the effect of cocoa flavanols on BP. • Few studies are of high quality, and several uncertainties that must be addressed before passing the health claim. • An authorized health claim should not be allowed, but a qualified health claim with a category "C" may be allowed. [ABSTRACT FROM AUTHOR]

• Development of guidelines for proficiency testing of botanical bioactive measurement. • Definition of system suitability acceptance criteria using multi-laboratory data. • Cocoa flavanol, a model for botanical bioactives quality testing harmonization. Flavanols and procyanidins are bioactives found in many foods including cocoa. The characterization of cocoa flavanols and procyanidins (CF) has historically been challenged by the lack of commercially available standards and weak chromatographic separation performances. The recent release of a reference material and the final authorization of an AOAC Official Method of Analysis (AOAC 2020.05) technically enables the standardization of CF testing. However, the practical implementation of CF testing for routine testing remains challenging for new laboratories, highlighting the need to define guidelines and acceptance criteria to verify normal method performance and user proficiency. This study leveraged the data generated from the recent multi-laboratory validation to define normal method performances. While the challenges associated with HILIC separation can be alleviated through a thorough system equilibration, monitoring system performances remains critical to routine operation and a laboratory's ability to generate reliable data. Guidelines for routine analysis were developed for system precision and bracketing standard recovery, as well as acceptance criteria for the analysis of the reference material. These guidelines not only complete a body of work that provide accessible, reliable, and robust CF analysis solution to research and quality laboratories, but also provide an example to facilitate the establishment and implementation of future international testing standards for botanical bioactives. [ABSTRACT FROM AUTHOR]

Abstract Cocoa shell (CS) is a co-product of the cocoa industry used mainly as fuel for boilers but with secondary applications as fertilizer and in animal feed. Although it is known that this material is rich in flavanols and alkaloids, to date, a study has not been conducted that has quantitatively identified these compounds in CS. Thus, the aim of this work was to characterize CS in terms of its composition, regarding catechin, epicatechin, procyanidin B2, caffeine and theobromine, and to evaluate the extraction kinetics of the total flavanols using pressurized liquid extraction (PLE) with absolute ethanol. For the determination of the extraction kinetic data, the DMAC method was used, while each compound was quantified using a UPLC-MS/MS analysis. The major compounds found were theobromine and epicatechin (mean values of 9.89 and 3.5 mg/g CS, respectively). PLE proved to be quite effective; the flavanols extraction yield was enhanced by increasing the temperature and extraction time however, high extraction times and temperatures degraded the procyanidins B2. Peleg's model applied to extraction data description provided a reasonable agreement with the experimental results, which allows their application in modeling and optimization of solid-liquid extraction of the total flavanols from cocoa bean shell. Graphical abstract Unlabelled Image Highlights • The major biocompounds found in CS were theobromine and epicatechin. • PLE was useful for the simultaneous extraction of alkaloids and flavanols from CS. • The PLE process was enhanced by increasing the temperature and extraction time. • High extraction times and temperatures led to degradation of procyanidin B2. • The Peleg's model was suitable to represent the experimental data and to determine the maximum extraction yield. [ABSTRACT FROM AUTHOR]

We evaluated the short-term effects of a flavanol-rich cocoa (FRC) on lipid profile and selected oxidative stress biomarkers such as oxidized low-density lipoprotein (oxLDL), glutathione (GSH), and F2-isoprostane. We also assessed whether FRC modulates plasma levels of polyunsaturated fatty acids (PUFA) in healthy individuals. The subjects (n=48) were randomly assigned to a low-cocoa group (1 g/d; ~55 mg flavanols) (n=16), middle-cocoa group (2 g/d; ~110 mg flavanols) (n=16), or a high-cocoa group (4 g/d; ~220 mg flavanols) (n=16). The samples were collected at baseline, at 1, 2, and 4 h post initial consumption of FRC, and after 4 weeks of FRC supplementation. The peak plasma concentration of (-)-epicatechin metabolites reached a maximum level (578±61 nM; P<.05) at 2 h after ingestion of FRC. After 4 weeks, total cholesterol (-12.37±6.63; P<.0001), triglycerides (-3.81±2.45; P<.0001), plasma LDL (-14.98±6.77; P<.0001), and oxLDL (-95.61±41.69; P<.0001) decreased in the high-cocoa group, compared with baseline. We also found that plasma high-density lipoprotein (HDL) (+3.37±2.06; P<.0001) concentrations increased significantly in the same group. Total GSH significantly increased in all FRC-treated groups (+209.73±146.8; P<.0001), while urinary F2-isoprostane levels decreased in the middle- (-0.73±0.16; P<.0001) and high-cocoa (-1.62±0.61; P<.0001) groups. At the end of the four-week study, a significant reduction of arachidonic acid (AA)/eicosapentaenoic acid (EPA) ratio was observed in the low-(-2.62±2.93; P=.003), middle- (-5.24±2.75; P<.0001) and high-cocoa (-7.76±4.96; P<.0001) groups, compared with baseline. Despite the small sample size used in this study, these data extend previous clinical and experimental studies, providing new insights into the health benefits of cocoa flavanols. [ABSTRACT FROM AUTHOR]

The cocoa varieties Criollo , Forastero and Trinitario , which have different organoleptic properties, quite often are mixed together in cocoa-based products. The objective of this work was to develop a rapid method for the simultaneous determination of polyphenols ( n = 35 ) and alkaloids ( n = 2 ) in cocoa-based products by using the high throughput advantages provided by the Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer (HRMS). The proposed procedure was optimized and validated in terms of selectivity and specificity (mass accuracy <5 ppm), sensitivity (instrumental limit of detection from 15 to 30 fg), linearity (r 2  > 0.990), accuracy (recovery range from 89.1 to 112.4%) and precision (relative standard deviation <10%). The method was applied to 80 cocoa-based samples. Alkaloids represented on average the 57.7% of assayed compounds. Flavanols and procyanidins were the most abundant polyphenols being quantified in a range between 261 and 668 mg/kg and from 127 to 405 mg/g, respectively. Mass equivalents but structural isomers bioflavonoids were characterized by using the HRMS-Orbitrap methodology here developed. [ABSTRACT FROM AUTHOR]

Roasting is an important cocoa processing step, but has been reported to reduce the polyphenol content in the beans. We investigated the impact of whole-bean roasting on the polyphenol content, aroma-related chemistry, and in vitro pancreatic lipase (PL) inhibitory activity of cocoa under a range of roasting conditions. Total phenolics, (−)-epicatechin, and proanthocyanidin (PAC) dimer – pentamer content was reduced by roasting. By contrast, roasting at 150 °C or greater increased the levels of catechin and PAC hexamers and heptamers. These compounds have greater PL inhibitory potency. Consistent with these changes in PAC composition and this previous data, we found that roasting at 170 °C time-dependently increased PL inhibitory activity. Cocoa aroma-related compounds increased with roasting above 100 °C, whereas deleterious sensory-related compounds formed at more severe temperatures. Our results indicate that cocoa roasting can be optimized to increase the content of larger PACs and anti-PL activity, while maintaining a favorable aroma profile. [ABSTRACT FROM AUTHOR]

In this study, we investigated how the acute physiological effects of cocoa flavanols might result in specific cognitive changes, in particular in temporal and spatial attention. To this end, we pre-registered and implemented a randomized, double-blind, placebo- and baseline-controlled crossover design. A sample of 48 university students participated in the study and each of them completed the experimental tasks in four conditions (baseline, placebo, low dose, and high-dose flavanol), administered in separate sessions with a 1-week washout interval. A rapid serial visual presentation task was used to test flavanol effects on temporal attention and integration, and a visual search task was similarly employed to investigate spatial attention. Results indicated that cocoa flavanols improved visual search efficiency, reflected by reduced reaction time. However, cocoa flavanols did not facilitate temporal attention nor integration, suggesting that flavanols may affect some aspects of attention, but not others. Potential underlying mechanisms are discussed. [ABSTRACT FROM AUTHOR]

Growing evidence support that insulin resistance may occur as a severe problem due to chronic energetic overfeeding and subsequent obesity. When an abundance of glucose and saturated fat enter the cell, impaired blood flow, hypoxia, inflammation and macrophage infiltration in obese adipose tissue may induce oxidative stress and insulin resistance. Excessive circulating saturated fatty acids ectopically accumulate in insulin-sensitive tissues and impair insulin action. In this context, excessive hepatic lipid accumulation may play a central, pathogenic role in insulin resistance. It is thought that dietary polyphenols may ameliorate obesity-related insulin resistance by attenuating inflammatory responses and oxidative stress. The most often occurring natural polyphenolic compounds are flavonoids. In this review, the possible mechanistic effect of flavonoid-rich natural products on insulin resistance-related metabolic pathways is discussed. Polyphenol intake can prevent high-fat-diet-induced insulin resistance via cell surface G protein-coupled estrogen receptors by upregulating the expression of related genes, and their pathways, which are responsible for the insulin sensitivity. [ABSTRACT FROM AUTHOR]

The potential health effects of cocoa flavanols are well described. Ranging from reducing risk of developing type 2 diabetes and cardiovascular disease at population levels, moderating disease risk factors including endothelial function and lipid metabolism in clinical trials and mechanistic studies in laboratory studies highlighting target tissues and pathways. However, translating these benefits into public health messages is problematic, due to the high energy and sugar content of many cocoa products, including chocolate. This review considered the role of sugar in cocoa products, what are its physiological effects on bioavailability and bioactivity? Considering, then how cocoa products can be reformulated to reduce sugar intake, and the likely effects on beneficial effects of cocoa flavanols and consumer preferences. Ultimately, although interesting physiological effects are seen with cocoa flavanols, their use as a disease-modifying commodities may be limited the effect such products may have within an individual's and populations overall dietary patterns. [ABSTRACT FROM AUTHOR]

Background Cocoa flavanols (CF) can stimulate vasodilation by improved nitric oxide (NO) synthesis and have antioxidant and anti-inflammatory capacities. This study aimed to examine whether acute CF intake can affect exercise-induced changes in antioxidant capacity, oxidative stress, inflammation and NO production, as well as exercise performance and recovery in well-trained cyclists. Methods Twelve well-trained male cyclists (mean ± SD age, VO2max: 30 ± 3 years, 63.0 ± 3.5 ml/kg/min) participated in this randomized, double-blind, cross over study. On 2 separate occasions, subjects performed two 30-min time trials 1.5 (TT1) and 3 (TT2) hours after CF (900 mg CF) or placebo (PL, 13 mg CF) intake, interposed by passive rest. Lactate, glucose, heartrate, rating of perceived exertion (RPE) and power output were measured during the TTs. Blood was drawn at baseline, before and after each TT and analyzed for epicatechin serum concentrations, trolox equivalent antioxidative capacity (TEAC), uric acid (UA), malonaldehyde (MDA), L-arginine/ADMA, citrulline, interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF)-α plasma concentrations. Relative changes in blood markers and pacing strategy during TT were analysed by repeated measured ANOVA. TT performance was compared between PL and CF by paired t-test. Results Epicatechin concentrations were increased by CF intake. Exercise-induced increase in TEAC/UA was improved by CF intake (F(1) = 5.57; p = .038) (post-TT1: PL: 113.34 ± 3.9%, CF: 117.64 ± 3.96%, post-TT2: PL: 108.59 ± 3.95%, CF: 123.72 ± 7.4% to baseline), while exercise-induced increases in MDA, IL-1 and IL-6 were not affected by CF intake. TNF-α was unaltered by exercise and by CF. Exercise-induced decreases in L-arginine/ADMA and increases in citrulline were not affected by CF intake. TT1 and TT2 performance and exercise-induced physiological changes were unaffected by CF intake. Conclusion Acute CF intake increased total antioxidant capacity in rest and during exercise, but did not affect exercise-induced lipid peroxidation, inflammation, nor NO production in healthy athletes. Acute CF intake did not improve TT performance and recovery. Trial registration ISRCTN32875, 21-11-2016, retrospectively registered.

It is well known that cocoa and dark chocolate possess polyphenols as major constituents whose dietary consumption has been associated to beneficial effects. In fact, cocoa and dark chocolate polyphenols exert antioxidant and anti-inflammatory activities switching on some important signaling pathways such as toll-like receptor 4/nuclear factor κB/signal transducer and activator of transcription. In particular, cocoa polyphenols induce release of nitric oxide (NO) through activation of endothelial NO synthase which, in turn, accounts for vasodilation and cardioprotective effects. In the light of the above described properties, a number of clinical trials based on the consumption of cocoa and dark chocolate have been conducted in healthy subjects as well as in different categories of patients, such as those affected by cardiovascular, neurological, intestinal, and metabolic pathologies. Even if data are not always concordant, modifications of biomarkers of disease are frequently associated to improvement of clinical manifestations. Quite interestingly, following cocoa and dark chocolate ingestion, cocoa polyphenols also modulate intestinal microbiota, thus leading to the growth of bacteria that trigger a tolerogenic anti-inflammatory pathway in the host. Finally, many evidences encourage the consumption of cocoa and dark chocolate by aged people for the recovery of the neurovascular unit.

Background: Cocoa flavanols (CF) can stimulate vasodilation by improved nitric oxide (NO) synthesis and have antioxidant and anti-inflammatory capacities. This study aimed to examine whether acute CF intake can affect exercise-induced changes in antioxidant capacity, oxidative stress, inflammation and NO production, as well as exercise performance and recovery in well-trained cyclists. Methods: Twelve well-trained male cyclists (mean ± SD age, VO2max: 30 ± 3 years, 63.0 ± 3.5 ml/kg/min) participated in this randomized, double-blind, cross over study. On 2 separate occasions, subjects performed two 30-min time trials 1.5 (TT1) and 3 (TT2) hours after CF (900 mg CF) or placebo (PL, 13 mg CF) intake, interposed by passive rest. Lactate, glucose, heartrate, rating of perceived exertion (RPE) and power output were measured during the TTs. Blood was drawn at baseline, before and after each TT and analyzed for epicatechin serum concentrations, trolox equivalent antioxidative capacity (TEAC), uric acid (UA), malonaldehyde (MDA), L-arginine/ADMA, citrulline, interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF)-α plasma concentrations. Relative changes in blood markers and pacing strategy during TT were analysed by repeated measured ANOVA. TT performance was compared between PL and CF by paired t-test. Results: Epicatechin concentrations were increased by CF intake. Exercise-induced increase in TEAC/UA was improved by CF intake (F(1) = 5.57; p =.038) (post-TT1: PL: 113.34 ± 3.9%, CF: 117.64 ± 3.96%, post-TT2: PL: 108.59 ± 3.95%, CF: 123.72 ± 7.4% to baseline), while exercise-induced increases in MDA, IL-1 and IL-6 were not affected by CF intake. TNF-α was unaltered by exercise and by CF. Exercise-induced decreases in L-arginine/ADMA and increases in citrulline were not affected by CF intake. TT1 and TT2 performance and exercise-induced physiological changes were unaffected by CF intake. Conclusion: Acute CF intake increased total antioxidant capacity in rest and during exercise, but did not affect exercise-induced lipid peroxidation, inflammation, nor NO production in healthy athletes. Acute CF intake did not improve TT performance and recovery. Trial registration: ISRCTN32875, 21-11-2016, retrospectively registered. [ABSTRACT FROM AUTHOR]