The most important studies on the properties of cocoa, phytochemical composition, intakes, allergies, etc.

Massaro M, Scoditti E, Carluccio MA, Kaltsatou A, Cicchella A. Effect of Cocoa Products and Its Polyphenolic Constituents on Exercise Performance and Exercise-Induced Muscle Damage and Inflammation: A Review of Clinical Trials. Nutrients. 2019 Jun 28;11(7):1471. doi: 10.3390/nu11071471.

Abstract. In recent years, the consumption of chocolate and, in particular, dark chocolate has been "rehabilitated" due to its high content of cocoa antioxidant polyphenols. Although it is recognized that regular exercise improves energy metabolism and muscle performance, excessive or unaccustomed exercise may induce cell damage and impair muscle function by triggering oxidative stress and tissue inflammation. The aim of this review was to revise the available data from literature on the effects of cocoa polyphenols on exercise-associated tissue damage and impairment of exercise performance. To this aim, PubMed and Web of Science databases were searched with the following keywords: "intervention studies", "cocoa polyphenols", "exercise training", "inflammation", "oxidative stress", and "exercise performance". We selected thirteen randomized clinical trials on cocoa ingestion that involved a total of 200 well-trained athletes. The retrieved data indicate that acute, sub-chronic, and chronic cocoa polyphenol intake may reduce exercise-induced oxidative stress but not inflammation, while mixed results are observed in terms of exercise performance and recovery. The interpretation of available results on the anti-oxidative and anti-inflammatory activities of cocoa polyphenols remains questionable, likely due to the variety of physiological networks involved. Further experimental studies are mandatory to clarify the role of cocoa polyphenol supplementation in exercise-mediated inflammation.

Gröne M, Sansone R, Höffken P, Horn P, Rodriguez-Mateos A, Schroeter H, Kelm M, Heiss C. Cocoa Flavanols Improve Endothelial Functional Integrity in Healthy Young and Elderly Subjects. J Agric Food Chem. 2020 Feb 19;68(7):1871-1876. doi: 10.1021/acs.jafc.9b02251.

Abstract. Cocoa flavanols (CFs) can improve flow-mediated dilation (FMD), blood pressure, and vascular stiffness in healthy subjects. Endothelial microparticles (EMPs) are markers of endothelial functional integrity, reflecting activation and injury. In plasma samples, we investigated whether age-dependent changes in circulating EMPs exist and whether CFs decrease EMPs in healthy humans. The concentrations of CD31+/41-, CD144+, and CD62e+ EMPs (flow cytometry) were increased in healthy elderly (n = 19) compared to young (n = 20) non-smokers. EMPs correlated with age, systolic blood pressure, and pulse wave velocity. CD31+/41- and CD62e+ EMPs inversely correlated with FMD. Following 2 weeks twice-daily CF consumption (450 mg), CD31+/41- and CD144+ EMPs decreased in both young and elderly subjects compared to the CF-free control. The EMP decrease inversely correlated with FMD improvements. Cardiovascular aging is associated with increased EMPs that can be modulated by dietary flavanols along with improvements in vascular function. This indicates that flavanol consumption can improve endothelial functional integrity in healthy humans.

Yoo, H., & Kim, H. S. (2019). Different Extraction Solvents and Sample Preparation Methods Affect the Antioxidant Activities and Total Phenol Contents of Theobroma Cacao Powder (P01-016-19). Current Developments in Nutrition, 3(Supplement_1), nzz028-P01.

Abstract. Objectives Cacao(Theobroma cacao) has been widely consumed since more than 4000 years ago which was regarded as sacred food. Cacao has been considered as a strong antioxidant due to its abundant phenolic and especially flavonoid contents. In this experiment, different extraction solvents and methods were used to analyze whether these factors affect the antioxidant activities and phytochemical compounds of cacao powder extracts. Methods Four different cacao extract samples were examined to identify their anti-oxidative activities; raw cacao powder extracted in 70% methanol solution(RM), raw cacao powder extracted in 80% ethanol solution(RE), freeze dried cacao powder extracted in 70% methanol solution(FM), freeze dried cacao powder extracted in 80% ethanol solution(FE). Total phenolic compound content was calculated based on gallic acid standard curve and total flavonoid content was calculated based on the standard curve of quercetin. ABTS scavenging activity was experimented with ABTS reagent diluted by PBS(pH7.4) to make the O.D. value 0.7 ± 0.02 at 734 nm before using 2.5 mM ABTS. The DPPH radical scavenging activity was assessed by 1 mM DPPH methanol solution. Results The contents of total phenol compounds were 45.4 mg GAE/g(RM), 37.3 mg GAE/g(RE), 50.6 mg GAE/g(FM), and 52.7 mg GAE/g(FE). The contents of total flavonoid compounds were 34.3 mg QE/g(RM), 29.7 mg QE/g(RE), 85.1 mg QE/g(FM), and 89.7 mg QE/g(FE). FM group showed the highest ABTS radical scavenging activity which is a marker of antioxidant activity but the other sample groups represented almost as high ABTS radical scavenging activity as FM group. RM group showed the highest DPPH radical scavenging activity which is also a marker of antioxidant activity. Conclusions The results show that different extract solvents and sample preparation methods affect DPPH radical scavenging activity, total phenol and flavonoid contents of cacao powder extracts. Therefore, appropriate methods should be applied according to the purpose of the study.

Rossin D, Barbosa-Pereira L, Iaia N, Testa G, Sottero B, Poli G, Zeppa G, Biasi F. A Dietary Mixture of Oxysterols Induces In Vitro Intestinal Inflammation through TLR2/4 Activation: The Protective Effect of Cocoa Bean Shells. Antioxidants (Basel). 2019 May 31;8(6):151. doi: 10.3390/antiox8060151.

Abstract. Background: Exaggerated Toll-like receptor (TLR)-mediated immune and inflammatory responses play a role in inflammatory bowel diseases. This report deals with the ability of a mixture of oxysterols widely present in cholesterol-rich foods to induce in vitro intestinal inflammation through TLR up-regulation. The anti-inflammatory action of four cocoa bean shell (CBS) extracts with different polyphenol content, was tested. Methods: Differentiated intestinal CaCo-2 cells were treated with a dietary oxysterol mixture (Oxy-mix) (60 µM). The expression and activation of TLR2 and TLR4, as well as the production of their downstream signaling effectors IL-8, IFNβ and TNFα were analyzed in the presence or absence of TLR antibodies. Honduras CBS extracts were characterized for their polyphenol contents; their anti-inflammatory action was analyzed in CaCo-2 cells treated with Oxy-mix. Results: Oxysterol-dependent TLR-2 and TLR4 over-expression and activation together with cytokine induction were abolished by blocking TLRs with specific antibodies. Polyphenol-rich CBS extracts consisting of high quantities of (-)-epicatechin and tannins also prevented TLR induction. Conclusions: TLR2 and TLR4 mainly contribute to inducing oxysterol-dependent intestinal inflammation. The fractionation method of CBS allowed the recovery of fractions rich in (-)-epicatechin and tannins able to counteract oxysterol-induced inflammation, thus highlighting the beneficial biological potential of specific CBS extracts.

Bhagat AR, Delgado AM, Issaoui M, Chammem N, Fiorino M, Pellerito A, Natalello S. Review of the Role of Fluid Dairy in Delivery of Polyphenolic Compounds in the Diet: Chocolate Milk, Coffee Beverages, Matcha Green Tea, and Beyond. J AOAC Int. 2019 Sep 1;102(5):1365-1372. doi: 10.5740/jaoacint.19-0129. 

Abstract. Dairy-based functional beverages have been a growing segment as consumer demands for health foods have shifted focus from simply enhancing lifespan to protecting health. Green tea is often limited in use because of poor bioavailability and disagreeable taste. However, milk is considered an ideal platform for the delivery of active polyphenolic compounds in green tea. Furthermore, the antioxidant enzymatic activity and antioxidant index of polyphenols in green tea have been known to be protected through interaction with dairy proteins inside the unstable intestinal environment. In addition, consumption of green tea infused with milk has been found to have a significant impact on reducing skin wrinkles and roughness in elderly subjects, through a decrease in lipid peroxidation and a concomitant reduction in oxidative stress. A similar affinity has been observed between antioxidants in coffee and milk proteins. Dark chocolate has been known to contain significant phenolic content and antioxidant activity. The activation of protein complex NF-κB, which is responsible for cell survival, was found to be significantly reduced upon consumption of cocoa with water, whereas consuming cocoa with milk had no effect on the bioavailability of the phenolic compounds in cocoa. The popularity of dairy as the source for polyphenol fortified beverages in the diet will be dictated by optimization of the technology for maximizing the bioavailability of the antioxidants.

Garcia JP, Santana A, Baruqui DL, Suraci N. The Cardiovascular effects of chocolate. Rev Cardiovasc Med. 2018 Dec 30;19(4):123-127. doi: 10.31083/j.rcm.2018.04.3187. 

Abstract. The antioxidants as polyphenols, especially flavanols present in cocoa, exert a favorable effect on endothelium vasodilation, modulate inflammatory markers, and decrease platelet aggregation, lipid oxidation and insulin resistance. Recent nutritional intervention trials and molecular studies demonstrate that consumption of cocoa, particularly rich in flavanols, is beneficial to promote cardiovascular health. This review describes the cardiovascular effects of chocolate.

Lopes JP, Kattan J, Doppelt A, Nowak-Węgrzyn A, Bunyavanich S. Not so sweet: True chocolate and cocoa allergy. J Allergy Clin Immunol Pract. 2019 Nov-Dec;7(8):2868-2871. doi: 10.1016/j.jaip.2019.04.023.

Abstract. Food allergy to chocolate and cocoa is considered possible, but there has been no report to date of challenge-proven, IgE-mediated allergy to chocolate or cocoa. Although population studies cite a prevalence of self-reported chocolate/cocoa allergy of 0.5 to 0.7% in select populations,1 self-reported food allergy is notoriously inaccurate, and objective testing was not used to corroborate chocolate or cocoa allergy status in these studies. 1 The majority of empirically defined chocolate and cocoa allergies are thought to be due to cross-contamination from common allergens, including peanut,2 tree nuts, or milk.3, 4 Here we report for the first time a case series of oral food challenge-confirmed, IgE-mediated chocolate/cocoa allergy.

Zięba K, Makarewicz-Wujec M, Kozłowska-Wojciechowska M. Cardioprotective Mechanisms of Cocoa. J Am Coll Nutr. 2019 Aug;38(6):564-575. doi: 10.1080/07315724.2018.1557087.

Abstract. Cardiovascular diseases are the main cause of deaths in highly developed countries. Dietetic interventions that involve recommendations for consumption of products with a confirmed health-improving action are an important aspect of prevention of cardiovascular diseases. Cocoa is an alimentary product with significant cardioprotective potential due to its high content of bioactive compounds. The aim of the present study was to review the most recent literature concerning the effectiveness and mechanisms of action of compounds contained in cocoa with regard to selected cardiovascular risk factors and cardiometabolic markers. Study results indicate that cocoa consumption, especially in the form of dark chocolate with high flavonoid content, may be a good strategy to diminish cardiovascular risk due to its beneficial effect on platelet aggregation, decreasing blood pressure, diminishing dyslipidemia, and decreasing blood plasma glucose concentration. Many studies have shown that cocoa-derived flavonoids have antioxidant and anti-inflammatory activity and also play a significant role in preventing insulin resistance. However, in order to completely confirm the potential cardiovascular benefits, it is necessary to conduct larger and longer studies, also with regard to potential dangers associated with long-term consumption of large amounts of flavonoids and determination of a safe and effective dose. Key teaching points Cocoa consumption may be a good strategy in diminishing cardiovascular risk. Beneficial effects on platelet aggregation, blood pressure, dyslipidemia, glycemia, as well as antioxidant and anti-inflammatory activity are observed. There is a need to conduct larger and longer studies to determine a safe and effective dose of cocoa flavonoids.

Cavarretta E, Peruzzi M, Del Vescovo R, Di Pilla F, Gobbi G, Serdoz A, Ferrara R, Schirone L, Sciarretta S, Nocella C, De Falco E, Schiavon S, Biondi-Zoccai G, Frati G, Carnevale R. Dark Chocolate Intake Positively Modulates Redox Status and Markers of Muscular Damage in Elite Football Athletes: A Randomized Controlled Study. Oxid Med Cell Longev. 2018 Nov 21;2018:4061901. doi: 10.1155/2018/4061901. 

 Abstract. Intensive physical exercise may cause increase oxidative stress and muscular injury in elite football athletes. The aim of this study was to exploit the effect of cocoa polyphenols on oxidative stress and muscular injuries induced by intensive physical exercise in elite football players. Oxidant/antioxidant status and markers of muscle damage were evaluated in 24 elite football players and 15 controls. Furthermore, the 24 elite football players were randomly assigned to either a dark chocolate (>85% cocoa) intake (n = 12) or a control group (n = 12) for 30 days in a randomized controlled trial. Oxidative stress, antioxidant status, and muscle damage were assessed at baseline and after 30 days of chocolate intake. Compared to controls, elite football players showed lower antioxidant power and higher oxidative stress paralleled by an increase in muscle damage markers. After 30 days of dark chocolate intake, an increased antioxidant power was found in elite athletes assuming dark chocolate. Moreover, a significant reduction in muscle damage markers (CK and LDH, p < 0.001) was observed. In the control group, no changes were observed with the exception of an increase of sNox2-dp, H2O2, and myoglobin. A simple linear regression analysis showed that sNox2-dp was associated with a significant increase in muscle damage biomarker release (p = 0.001). An in vitro study also confirmed that polyphenol extracts significantly decreased oxidative stress in murine myoblast cell line C2C12-derived. These results indicate that polyphenol-rich nutrient supplementation by means of dark chocolate positively modulates redox status and reduced exercise-induced muscular injury biomarkers in elite football athletes. This trial is registered with NCT03288623.

Cocoa is derived from a plant called Theobroma cocoa that is born in South America and from which the seeds of the fruits are obtained.

These seeds undergo a fairly complex processing divided into several stages:

• Fermentation. The seeds are collected and piled into baskets and wooden vats and then exposed to a temperature of between 40 and 50 degrees. At this stage the sugars turn into acids and eliminate what is left of the pulp and lower the bitter taste of the seeds.
• Drying. Exposure to the sun or through jets of hot air makes the product lose half its weight.
• Toasting. The temperature is kept around 100/150 degrees.
  Various shelling, crushing and cleaning procedures. 
• Grinding. At this stage you mix different qualities of cocoa to get the desired taste and quality.

At the end of the processing you get a dough that will serve for all subsequent processing, but especially for the most important the :

Cocoa paste or cocoa mass

From this pasta that results in a very bitter taste, they are made with various processes:

• Cocoa butter : obtained by pressure and heat
• Cocoa powder : with fine grinding until you get a soluble powder in water.
• Chocolate : with complex and articulated processing.

Cocoa (Theobroma cacao L., Malvaceae)

Cocoa encompasses seeds (beans/nibs) and derivatives obtained from processing Theobroma cacao: cocoa liquor (mass), cocoa butter, and powders (natural or Dutch-processed/alkalized). It is used as a flavoring, functional, and lightly coloring ingredient in foods and beverages; in cosmetics as an emollient (butter), gentle exfoliant (powders), and antioxidant extract.

Caloric Value (Per 100 g Of Product)

Roasted beans/nibs: ~570–620 kcal/100 g.
Cocoa liquor (mass): ~580–650 kcal/100 g.
Cocoa powder 10–12% fat (natural/Dutch): ~220–260 kcal/100 g.
Cocoa powder 20–22% fat: ~360–450 kcal/100 g.
Cocoa butter: ~880–900 kcal/100 g.
Unsweetened cocoa beverage (ready-to-drink): ~0–15 kcal/100 g (depends on dissolved solids).

At practical use levels, energy contribution depends on form and recipe; sugar additions markedly raise calories.

Key Constituents

Polyphenols: flavanols (catechin, epicatechin) and B-type procyanidins; phenolic acids. They contribute color and bitterness/astringency and show in-vitro antioxidant activity (TPC as a global indicator; profile via HPLC).
Xanthine alkaloids: theobromine (major) and trace caffeine; responsible for bitter note and mild stimulation.
Lipids (derivatives/residual): cocoa butter with typical fatty acids—stearic (SFA — saturated fatty acids; high oxidative stability, dietary balance advisable), oleic (MUFA — monounsaturated fatty acids; often favorable for oxidative stability), palmitic (SFA), and small linoleic (PUFA — polyunsaturated fatty acids; functional but more oxidation-prone).
Dietary fiber: high in powders and nibs.
Minerals: magnesium, potassium, iron, copper (variable); regulated metals (e.g., cadmium) must be monitored.
Volatiles: pyrazines, aldehydes, esters formed through fermentation/roasting.
Analytical markers: TPC, flavanol/xanthine profile by HPLC, moisture/aw, color (Lab*), pH; for powders, particle size and fat content.

Production Process

Harvest & fermentation: pod opening; pulp/seed fermentation (2–7 days) to develop aroma precursors.
Drying: moisture reduction for stability and storability.
Roasting: aroma development and microbial reduction.
Winnowing (dehulling): shell/nib separation.
Grinding: production of cocoa liquor; optional pressing to separate cocoa butter and press cake, then milled into powder.
Alkalization (Dutch process): carbonate treatment (pH ↑, darker color, lower acidity; potential polyphenol reduction).
Quality controls: moisture/aw, pH, total fat, TPC/HPLC, mycotoxins (e.g., OTA), metals (e.g., Cd), pesticides, PAHs; packaging per GMP/HACCP.

Sensory And Technological Properties

Aroma/color: roasted-cocoa, nutty, caramelized notes; Dutch powders are darker and less acidic.
Functionality: brown coloration, in-vitro antioxidant activity, contribution to body and texture; cocoa butter provides structure and gloss in coatings/creams.
Compatibility: in protein beverages, polyphenol–protein complexes can cause haze/precipitation; alkalization improves dispersibility but may reduce TPC.

Food Applications

Chocolate & fillings: liquor/butter per standards of identity.
Beverages & RTD: 1–4% powder (higher for Dutch); stabilize with hydrocolloids.
Bakery: 1–6% in flours/doughs; consider pH for chemical leavening.
Dairy/ice cream: 1.5–5% powder; contributes color, bitterness, and body.
Cereals/snacks/spreads: dosage per sensory target and fat/water binding.

Nutrition And Health

Cocoa provides polyphenols with in-vitro antioxidant activity; in foods, health claims require authorization. Theobromine and caffeine warrant attention in sensitive individuals; the product is not suitable for pets. Nutritional impact depends on context (added sugars, fat level).

Quality And Specifications (Typical Topics)

Key parameters: moisture (often ≤8%), aw, total fat (10–12% or 20–22% in powders), pH (natural ~5.2–6.0; Dutch ~6.8–8.1), color Lab*, particle size.
Composition: TPC/HPLC (flavanols), theobromine/caffeine, ash.
Safety: mycotoxins (OTA), metals (Cd/Pb) within limits, pesticides, PAHs compliant; microbiology to spec.
Sensory: free from moldy, rancid, overly smoky notes.
Traceability & hygiene: GMP/HACCP compliance.

Storage And Shelf Life

Protect from humidity, light, and oxygen (DO); use low-permeability barrier packs.
Powders: control RH/aw to avoid caking and aroma loss; reseal well.
Butter/liquor: limit oxidation (cool temperature, low light, low oxygen); avoid thermal shock.
Apply FIFO rotation.

Allergens And Safety

Cocoa is not a major allergen, yet cross-contact (milk/tree nuts) can occur in facilities. Verify labeling where required for theobromine/caffeine and compliance with mycotoxin/metal limits.

INCI Functions In Cosmetics

Typical entries: Theobroma Cacao (Cocoa) Seed Butter; Theobroma Cacao Extract; Theobroma Cacao (Cocoa) Seed Powder.
Roles: emollient (butter), antioxidant and skin conditioning (extracts), gentle physical exfoliant (powders), masking.

Troubleshooting

Excess bitterness/astringency: high dose or tannin-rich fraction → reduce dose, choose “softer” powders, balance with fats/sugars, optimize pH.
Haze/precipitates in beverages: polyphenol–protein or Ca/Mg complexes → clarification, fine filtration, mild chelants; check water hardness.
Underpowered color: low-pH natural powder → consider Dutch powder (higher pH) or higher dose; note possible TPC decrease.
Over-limit Cd/OTA: non-compliant raw material → change origin/supplier, tighten intake screening, reinforce CCP.

Sustainability And Supply Chain

Valorization of co-products (shells for extracts/fiber, biochar/compost), effluent management to BOD/COD targets, recyclable packaging, and controlled temperature/humidity logistics reduce environmental impact. Provenance traceability and responsible farming practices support sustainability.

Conclusion

Cocoa offers a complex aroma profile, a useful polyphenolic matrix, and broad technological functionality. Performance and stability depend on raw-material quality, pH/process choices (alkalization), protection from humidity/oxygen, and analytical standardization.

Mini-Glossary

TPC — Total phenolic content: Folin–Ciocalteu global, non-specific phenolic indicator.
HPLC — High-performance liquid chromatography: quantitative analysis of flavanols and xanthines.
SFA — Saturated fatty acids (e.g., stearic/palmitic): high oxidative stability; dietary moderation advisable.
MUFA — Monounsaturated fatty acids (e.g., oleic): often favorable for lipid profile and stability.
PUFA — Polyunsaturated fatty acids (e.g., linoleic): functional but more oxidation-prone; protect accordingly.
DO — Dissolved oxygen: lowering it limits oxidation and aroma loss.
RH — Relative humidity: control to prevent caking and degradation.
aw — Water activity: “free” water fraction tied to stability and microbiology.
GMP/HACCP — Good manufacturing practice / Hazard analysis and critical control points: preventive quality systems with defined CCP.
BOD/COD — Biochemical/chemical oxygen demand: effluent organic-load indicators.
PAHs — Polycyclic aromatic hydrocarbons: regulated contaminants in roasted matrices.
CCP — Critical control point: step where a control prevents/eliminates/reduces a hazard.

Cocoa studies

References__________________________________________________________________________

Montagna MT, Diella G, Triggiano F, Caponio GR, De Giglio O, Caggiano G, Di Ciaula A, Portincasa P. Chocolate, "Food of the Gods": History, Science, and Human Health. Int J Environ Res Public Health. 2019 Dec 6;16(24):4960. doi: 10.3390/ijerph16244960. 

Abstract. Chocolate is well known for its fine flavor, and its history began in ancient times, when the Maya considered chocolate (a cocoa drink prepared with hot water) the "Food of the Gods". The food industry produces many different types of chocolate: in recent years, dark chocolate, in particular, has gained great popularity. Interest in chocolate has grown, owing to its physiological and potential health effects, such as regulation of blood pressure, insulin levels, vascular functions, oxidation processes, prebiotic effects, glucose homeostasis, and lipid metabolism. However, further translational and epidemiologic studies are needed to confirm available results and to evaluate other possible effects related to the consumption of cocoa and chocolate, verifying in humans the effects hitherto demonstrated only in vitro, and suggesting how best to consume (in terms of dose, mode, and time) chocolate in the daily diet.

Ferri C, Desideri G, Ferri L, Proietti I, Di Agostino S, Martella L, Mai F, Di Giosia P, Grassi D. Cocoa, blood pressure, and cardiovascular health. J Agric Food Chem. 2015 Nov 18;63(45):9901-9. doi: 10.1021/acs.jafc.5b01064. 

Abstract. High blood pressure is an important risk factor for cardiovascular disease and cardiovascular events worldwide. Clinical and epidemiological studies suggest that cocoa-rich products reduce the risk of cardiovascular disease. According to this, cocoa has a high content in polyphenols, especially flavanols. Flavanols have been described to exert favorable effects on endothelium-derived vasodilation via the stimulation of nitric oxide-synthase, the increased availability of l-arginine, and the decreased degradation of NO. Cocoa may also have a beneficial effect by protecting against oxidative stress alterations and via decreased platelet aggregation, decreased lipid oxidation, and insulin resistance. These effects are associated with a decrease of blood pressure and a favorable trend toward a reduction in cardiovascular events and strokes. Previous meta-analyses have shown that cocoa-rich foods may reduce blood pressure. Long-term trials investigating the effect of cocoa products are needed to determine whether or not blood pressure is reduced on a chronic basis by daily ingestion of cocoa. Furthermore, long-term trials investigating the effect of cocoa on clinical outcomes are also needed to assess whether cocoa has an effect on cardiovascular events. A 3 mmHg systolic blood pressure reduction has been estimated to decrease the risk of cardiovascular and all-cause mortality. This paper summarizes new findings concerning cocoa effects on blood pressure and cardiovascular health, focusing on putative mechanisms of action and "nutraceutical " viewpoints.

Jean-Marie E, Jiang W, Bereau D, Robinson JC. Theobroma cacao and Theobroma grandiflorum: Botany, Composition and Pharmacological Activities of Pods and Seeds. Foods. 2022 Dec 8;11(24):3966. doi: 10.3390/foods11243966. 

Abstract. Cocoa and cupuassu are evergreen Amazonian trees belonging to the genus Theobroma, with morphologically distinct fruits, including pods and beans. These beans are generally used for agri-food and cosmetics and have high fat and carbohydrates contents. The beans also contain interesting bioactive compounds, among which are polyphenols and methylxanthines thought to be responsible for various health benefits such as protective abilities against cardiovascular and neurodegenerative disorders and other metabolic disorders such as obesity and diabetes. Although these pods represent 50-80% of the whole fruit and provide a rich source of proteins, they are regularly eliminated during the cocoa and cupuassu transformation process. The purpose of this work is to provide an overview of recent research on cocoa and cupuassu pods and beans, with emphasis on their chemical composition, bioavailability, and pharmacological properties. According to the literature, pods and beans from cocoa and cupuassu are promising ecological and healthy resources.

Tan TYC, Lim XY, Yeo JHH, Lee SWH, Lai NM. The Health Effects of Chocolate and Cocoa: A Systematic Review. Nutrients. 2021 Aug 24;13(9):2909. doi: 10.3390/nu13092909. 

Abstract. Chocolate has a history of human consumption tracing back to 400 AD and is rich in polyphenols such as catechins, anthocyanidins, and pro anthocyanidins. As chocolate and cocoa product consumption, along with interest in them as functional foods, increases worldwide, there is a need to systematically and critically appraise the available clinical evidence on their health effects. A systematic search was conducted on electronic databases such as MEDLINE, EMBASE, and Cochrane Central Register of Controlled Trials (CENTRAL) using a search strategy and keywords. Among the many health effects assessed on several outcomes (including skin, cardiovascular, anthropometric, cognitive, and quality of life), we found that compared to controls, chocolate or cocoa product consumption significantly improved lipid profiles (triglycerides), while the effects of chocolate on all other outcome parameters were not significantly different. In conclusion, low-to-moderate-quality evidence with short duration of research (majority 4-6 weeks) showed no significant difference between the effects of chocolate and control groups on parameters related to skin, blood pressure, lipid profile, cognitive function, anthropometry, blood glucose, and quality of life regardless of form, dose, and duration among healthy individuals. It was generally well accepted by study subjects, with gastrointestinal disturbances and unpalatability being the most reported concerns.

Schwan RF, Wheals AE. The microbiology of cocoa fermentation and its role in chocolate quality. Crit Rev Food Sci Nutr. 2004;44(4):205-21. doi: 10.1080/10408690490464104. 

Abstract. The first stage of chocolate production consists of a natural, seven-day microbial fermentation of the pectinaceous pulp surrounding beans of the tree Theobroma cacao. There is a microbial succession of a wide range of yeasts, lactic-acid, and acetic-acid bacteria during which high temperatures of up to 50 degrees C and microbial products, such as ethanol, lactic acid, and acetic acid, kill the beans and cause production of flavor precursors. Over-fermentation leads to a rise in bacilli and filamentous fungi that can cause off-flavors. The physiological roles of the predominant micro-organisms are now reasonably well understood and the crucial importance of a well-ordered microbial succession in cocoa aroma has been established. It has been possible to use a synthetic microbial cocktail inoculum of just 5 species, including members of the 3 principal groups, to mimic the natural fermentation process and yield good quality chocolate. Reduction of the amount of pectin by physical or mechanical means can also lead to an improved fermentation in reduced time and the juice can be used as a high-value byproduct. To improve the quality of the processed beans, more research is needed on pectinase production by yeasts, better depulping, fermenter design, and the use of starter cultures.

Cárdenas A, Mojica L, Coronado-Cáceres L, Castillo-Herrera GA. Unlocking the Alkaloid Biological Potential of Chili Pepper (Capsicum spp.), Cacao (Theobroma cacao L.), and Coffee (Coffea spp.) Byproducts: Characterization, Non-Conventional Extraction, Applications, and Future Perspectives. Molecules. 2025 Sep 18;30(18):3795. doi: 10.3390/molecules30183795. 

Abstract. Chili peppers (Capsicum spp.), cacao (Theobroma cacao L.), and coffee (Coffea spp.) are important crops worldwide. Nearly 35%, 80%, and 45% of the respective fruits are underutilized or discarded, representing a considerable economic loss. This work reviews and analyzes the environmental factors that influence the concentration of the main alkaloids in these crops, including capsaicin, theobromine, and caffeine. Their reported anti-inflammatory, cardioprotective, neuroprotective, and cytotoxic properties are also reviewed. This work explores strategies for the revalorization of these crops, comparing alkaloid extraction methods that use non-conventional techniques, including supercritical fluid extraction (SFE), ultrasound-assisted extraction (UAE), high-pressure and -temperature extraction (HPTE), pressurized liquid extraction (PLE), pressurized hot water extraction (PHWE), enzyme-assisted extraction (EAE), and pulsed electric field-assisted extraction (PEFAE), and their combination to enhance the recovery of capsaicin, theobromine, and caffeine, leading to sustainable and innovative uses of these crops' byproducts. Capsaicin, theobromine, and caffeine alkaloids are promising ingredients for the development of functional foods, cosmeceuticals, and pharmaceuticals.

Díaz-Valderrama JR, Leiva-Espinoza ST, Aime MC. The History of Cacao and Its Diseases in the Americas. Phytopathology. 2020 Oct;110(10):1604-1619. doi: 10.1094/PHYTO-05-20-0178-RVW. 

Abstract. Cacao is a commodity crop from the tropics cultivated by about 6 million smallholder farmers. The tree, Theobroma cacao, originated in the Upper Amazon where it was domesticated ca. 5450 to 5300 B.P. From this center of origin, cacao was dispersed and cultivated in Mesoamerica as early as 3800 to 3000 B.P. After the European conquest of the Americas (the 1500s), cacao cultivation intensified in several loci, primarily Mesoamerica, Trinidad, Venezuela, and Ecuador. It was during the colonial period that cacao diseases began emerging as threats to production. One early example is the collapse of the cacao industry in Trinidad in the 1720s, attributed to an unknown disease referred to as the "blast". Trinidad would resurface as a production center due to the discovery of the Trinitario genetic group, which is still widely used in breeding programs around the world. However, a resurgence of diseases like frosty pod rot during the republican period (the late 1800s and early 1900s) had profound impacts on other centers of Latin American production, especially in Venezuela and Ecuador, shifting the focus of cacao production southward, to Bahia, Brazil. Production in Bahia was, in turn, dramatically curtailed by the introduction of witches' broom disease in the late 1980s. Today, most of the world's cacao production occurs in West Africa and parts of Asia, where the primary Latin American diseases have not yet spread. In this review, we discuss the history of cacao cultivation in the Americas and how that history has been shaped by the emergence of diseases.

Martin MÁ, Ramos S. Impact of cocoa flavanols on human health. Food Chem Toxicol. 2021 May;151:112121. doi: 10.1016/j.fct.2021.112121. Epub 2021 Mar 13. PMID: 33722594.

Kerimi A, Williamson G. The cardiovascular benefits of dark chocolate. Vascul Pharmacol. 2015 Aug;71:11-5. doi: 10.1016/j.vph.2015.05.011. Epub 2015 May 27. PMID: 26026398.