Preservation of Cocoa Antioxidant Activity, Total Polyphenols, Flavan-3-ols, and Procyanidin Content in Foods Prepared with Cocoa Powder

ABSTRACT:  Little is known about the effects of common cooking processes on cocoa flavanols. Antioxidant activity, total polyphenols (TP), flavanol monomers, and procyanidin oligomers were determined in chocolate frosting, a hot cocoa drink, chocolate cookies, and chocolate cake made with natural cocoa powder. Recoveries of antioxidant activity, TP, flavanol monomers, and procyanidins ranged from 86% to over 100% in the chocolate frosting, hot cocoa drink, and chocolate cookies. Losses were greatest in the chocolate cake with recoveries ranging from 5% for epicatechin to 54% for antioxidant activity. The causes of losses in baked chocolate cakes were investigated by exchanging baking soda with baking powder or combinations of the 2 leavening agents. Use of baking soda as a leavening agent was associated with increased pH and darkening color of cakes. Losses of antioxidant activity, TP, flavanol monomers, and procyanidins were associated with an increased extractable pH of the baked cakes. Chocolate cakes made with baking powder for leavening resulted in an average extractable pH of 6.2 with essentially complete retention of antioxidant activity and flavanol content, but with reduced cake heights and lighter cake color. Commercially available chocolate cake mixes had final pHs above 8.3 and contained no detectable monomeric flavanols after baking. These results suggest that baking soda causes an increase in pH and subsequent destruction of flavanol compounds and antioxidant activity. Use of an appropriate leavening agent to moderate the final cake pH to approximately 7.25 or less results in both good leavening and preservation of cocoa flavanols and procyanidins.     

Factors influencing quality variation in cocoa (Theobroma cacao) bean flavour profile — A review

This review examined the factors that influence flavour volatiles of cocoa beans and the volume of work that needs to be done on these factors and their impact on the flavour volatiles of commercial cocoa beans. Cocoa bean flavour is one of the most important quality attributes as flavour is central to acceptability of cocoa beans and cocoa products such as chocolate. The complex composition of cocoa bean flavour depends on bean genotype, postharvest treatments such as pulp pre-conditioning, fermentation and drying, industrial processes such as roasting as well as the type of soil and age of cocoa tree. The bean genotype determines the chemical composition of the bean, specifically the contents of bean storage proteins, polysaccharides, and polyphenols. This determines the quantities and type of precursors formed during fermentation and drying processes leading to flavour formation, hence, influencing both flavour type and intensity. Cocoa bean fermentation and drying result in the breakdown of the storage proteins by endogenous proteases into amino acids and short chain oligopeptides while the polysaccharides are also degraded by invertase to glucose and fructose. The amino acids, oligopeptides, glucose and fructose react with each other during the roasting process to produce the typical cocoa flavour volatiles. Polyphenols are also oxidized by polyphenol oxidase during fermentation and drying which reduce the astringency and bitterness of the beans, thus, enhancing the flavour of cocoa beans. However, the extent to which other factors such as age of the cocoa tree and soil chemical compositions influence the formation of flavour precursors and their relationships with final flavour quality remains unclear. With increasing demand for sustainable production of high quality cocoa beans, greater understanding of factors contributing to the variations in flavour character would have significant commercial implications.     

Tocopherols in cocoa butter obtained from cocoa bean roasted in different forms and under various process parameters

Obtaining good quality chocolate strongly depends on raw material, i.e. cocoa beans. The processing of cocoa beans consists of some important steps, including fermentation, drying fermented beans and roasting. Traditionally roasting is performed on whole beans but currently, roasting crushed cocoa beans or cocoa liquor becomes more and more popular. Many biologically active compounds may be found in the cocoa beans, including tocopherols. This work investigates the influence of the constant or variable roasting process parameters (temperature, velocity and relative humidity of roasting air) on the tocopherol concentration in cocoa butter (CB) extracted from cocoa beans originating from Togo and roasted in two different forms, namely as whole and crushed beans. Whole cocoa beans were roasted to a 2% moisture content and crushed cocoa beans were firstly partially dried which further enables easier dehusking, then ground, dehusked and roasted until their humidity decreased to around 2%. Roasting resulted in lowering the content of individual tocopherols in analyzed material. The degree of degradation of tocopherols in CBs was different, depending on the form of roasted beans from which these CBs were extracted. Higher concentrations were determined in CBs extracted from beans roasted in the form of crushed samples comparing to CBs obtained from beans roasted as a whole. The study investigates different roasting parameters of crushed beans, none of which drastically lowered the concentration of tocopherols in extracted CBs. Their concentration in CBs extracted from whole beans was, on the other hand, influenced by roasting air parameters. In case of whole beans roasted under constant parameters, application of 150 °C proved to be more favorable than roasting at 135 °C, as well as application of “dry” air and 1 m/s roasting air velocity. Discussing the variable roasting process parameters, in case of applied roasting temperature it is more favorable to change it from 150 °C to 135 °C, than the other way round. Changing the relative humidity of roasting air from 5 to 0.3%, lower degradation of tocopherols in CB occurs when the process is conducted at 150 °C. It may be further concluded that a direct dependence between the velocity of roasting air varied during the process and the concentration of tocopherols in extracted CB may not be indicated. In conclusion, it is stated that the temperature of the air applied during the roasting process has the decisive influence on the tocopherol content in CBs extracted from cocoa beans subjected to the process.     

Influence of roasting conditions on fatty acid composition and oxidative changes of cocoa butter extracted from cocoa bean of Forastero variety cultivated in Togo

Cocoa butter is responsible for physico-chemical, rheological and sensory properties of chocolates. Forastero cocoa beans are rich in cocoa butter. Roasting of cocoa beans is an essential step of their processing, giving rise to many desirable transformations. On the other hand, it causes changes in certain valuable ingredients including also cocoa butter. This article studies the influence of roasting conditions, i.e. temperature, humidity and flow velocity of roasting air, on the fatty acid composition, peroxide value and content of conjugated dienes and trienes in cocoa butter extracted from cocoa beans. Generally, in most cases roasting did not increase the peroxide value. In the constant conditions due to obtained results of PV, CD and CT values it is recommended to roast cocoa beans at the temperature of 150 °C, 1 m/s air velocity and 5% relative humidity. In the variable conditions lower values of PV, CD and CT were achieved when cocoa beans were roasted in variants with changing temperature from 150 °C to 135 °C or with changing air velocity from 1 to 0.5 m/s. Generally, the roasting process substantially did not change the content of each fatty acid, regardless of the applied conditions.     

Antioxidant Properties of Cocoa Beans (Theobroma cacao L.): Influence of Cultivar and Roasting Conditions

Effects of various roasting conditions on antioxidant properties of five Theobroma cacao L. varieties were investigated. The cocoa beans were roasted at four different temperatures (110–150°C) and three different air humidities (0.3–5.0%). The raw cocoa beans were characterized by high antioxidant activities. The antioxidant properties of the roasted cocoa beans varied markedly among the analyzed cultivars and geographical regions and were affected by roasting conditions. Generally, cocoa beans of the cv. Forastero from Brazil exhibited higher total phenolic content, free radical scavenging activity, and metal chelating ability than samples of the other analyzed cocoa varieties. Roasting at 110°C caused negligible changes in total phenolics content and antioxidant activity of cocoa beans, while almost all samples tended to have lower antioxidant potential when roasting temperature increased. The air humidity used in roasting did not affect the total phenolics content and antioxidant activity for lowest roasting temperature (110°C). Moreover, the obtained results revealed that thermal processing at the higher temperatures and elevated air humidity resulted in the higher antioxidant capacities. It was also found that the ferrous ion chelating activity of cocoa beans increased with the roasting temperature (in the range from 110 to 150°C), with the exception of cv. Trinitario from Papua New Guinea. The data showed that roasting at lower temperatures with humid air are more favorable in terms of preserving the bioactivity of roasted cocoa beans.     

发酵和焙烤对可可豆多酚、黄酮和风味品质的影响

本实验采用高效液相色谱仪（high performance liquid chromatography,HPLC）、色差仪、电子鼻分别检测未发酵、发酵和焙烤海南可可豆的总酚含量、总黄酮含量、色度和可可豆风味差异。结果表明：未经焙烤的未发酵豆总酚（464.03 mg/10 g）和总黄酮（126.86 mg/10 g）含量明显高于发酵豆总酚（211.86 mg/10 g）和总黄酮（61.98 mg/10 g）含量。105～145 ℃焙烤30 min,未发酵豆总酚和总黄酮含量分别为419.5～129.8 mg/10 g和77.8～16.8 mg/10 g;发酵豆总酚和总黄酮含量分别为182.53～86.25 mg/10 g和34.7～7.0 mg/10 g。其中,125 ℃焙烤20～40 min,未发酵豆总酚和黄酮含量分别为353.74～289.45 mg/10 g和42.86～32.20 mg/10 g;发酵豆总酚和黄酮含量分别为152.08～123.55 mg/10 g和25.12～21.14 mg/10 g。在105～145 ℃的温度焙烤下没食子酸含量变化显著。可可豆色度值的范围：L*值集中在40.0～47.0之间,a*值集中在5.0～6.8之间,b*值集中在4.0～8.5之间。未发酵和发酵可可豆之间,以及不同温度焙烤可可豆之间的电子感官风味分析结果差异较大。     

Quantification of 3-aminopropionamide in cocoa, coffee and cereal products

Based on recent results confirming 3-aminopropionamide (3-APA) as a very effective precursor of acrylamide in the absence of further “catalysts”, this compound was quantified for the first time in cocoa masses, cocoa beans, coffee and cereal products by LC–MS–MS after derivatisation with dansyl chloride. Cocoa masses contained >3000 μg/kg of 3-APA, but varied significantly in its concentration. For the quantification of acrylamide (AA) in cocoa and coffee, an improved isolation procedure using charcoal was developed. In various samples of unroasted and roasted cocoa beans, the concentrations of AA were by a factor of >5 lower than those of 3-APA, but the concentrations of 3-APA and AA were more closely correlated as compared to the concentrations of AA and Asparagine. Experiments on authentic cocoa beans from Ghana and Sulawesi indicated that the thermal generation of 3-APA during roasting was much more pronounced as compared to its biochemical formation. By administering fermented cocoa beans with [¹³C₄ ¹⁵N₂]-asparagine before roasting, 3-APA was confirmed as transient intermediate in AA formation during cocoa roasting. Among the cereal products analysed, in particular popcorn contained quite high amounts of 3-APA, which were also well correlated with the AA concentration. Contrary, in coffee products, 3-APA was always lower than AA.     

Application of various methods for determination of the color of cocoa beans roasted under variable process parameters

Cocoa beans of Forastero variety from Togo were subjected to roasting under either constant or variable process parameters. The variable process parameters were roasting air flow rate, temperature and relative humidity. The color of roasted cocoa beans was determined by pigment extraction under various conditions followed by either their spectrophotometric assays or CIE L*a*b measurements. Also the Maillard compounds index and total polyphenols content correlated with progress of the browning of beans were determined. It was found that an increase in the roasting air relative humidity stimulated formation of brown pigments, while elevated temperature caused worsening of color parameters of roasted cocoa beans. The most suitable method of color characterization of roasted cocoa beans was found to be pigments extraction combined with either separation of their fractions or CIE L*a*b measurements. These assays revealed that cocoa beans roasted under variable roasting air flow rate were characterized by improved color parameters. The relatively simple and inexpensive CIE L*a*b measurements ensured fast analysis of color parameters, while total polyphenols in roasted cocoa beans were quickly estimated by using F–C reagent. Furthermore, quantification of melanoid pigments in roasted cocoa beans can be based on determination of the index of nonenzymatic browning products content, which is relatively simple and inexpensive.     

Determining degree of roasting in cocoa beans by artificial neural network (ANN)‐based electronic nose system and gas chromatography/mass spectrometry (GC/MS)

BACKGROUND

Roasting is a critical step in chocolate processing, where moisture content is decreased and unique flavors and texture are developed. The determination of the degree of roasting in cocoa beans is important to ensure the quality of chocolate. Determining the degree of roasting relies on human specialists or sophisticated chemical analyses that are inaccessible to small manufacturers and farmers. In this study, an electronic nose system was constructed consisting of an array of gas sensors and used to detect volatiles emanating from cocoa beans roasted for 0, 20, 30 and 40 min. The several signals were used to train a three‐layer artificial neural network (ANN). Headspace samples were also analyzed by gas chromatography/mass spectrometry (GC/MS), with 23 select volatiles used to train a separate ANN.

RESULTS

Both ANNs were used to predict the degree of roasting of cocoa beans. The electronic nose had a prediction accuracy of 94.4% using signals from sensors TGS 813, 826, 822, 830, 830, 2620, 2602 and 2610. In comparison, the GC/MS predicted the degree of roasting with an accuracy of 95.8%.

CONCLUSION

The electronic nose system is able to predict the extent of roasting, as well as a more sophisticated approach using GC/MS. © 2018 Society of Chemical Industry     

Identification of novel cocoa flavonoids from raw fermented cocoa beans by HPLC–MSn

Using a detailed HPLC–ESI-MS analysis including both high resolution and tandem MS analysis, a series of fourteen novel flavonoids were identified from alcoholic extracts of raw fermented cocoa beans from Cameroon. The novel compounds include a series of isomeric hexosides of flavan-3-ols and novel proanthocyanidine dimers and trimers and a sulfated flavan-3-ol. Additionally a jasmonic acid sulfate derivative was identified in the cocoa bean extracts as one of the major metabolites produced under fermentation stress. Therefore, the chosen HPLC–ESI-MS analysis method revealed a greater complexity of the cocoa bean polyphenolic extract than previously reported and allowed individual assignment of novel compounds.     

Feasibility study on the use of Fourier transform near-infrared spectroscopy together with chemometrics to discriminate and quantify adulteration in cocoa beans

Fourier transform near-infrared (FT-NIR) spectroscopy combined with Support Vector Machine (SVM) and synergy interval partial least square (Si-PLS) was attempted in this study for cocoa bean authentication. SVM was used to develop an identification model to discriminate between fermented cocoa beans (FC), unfermented cocoa beans (UFC) and adulterated cocoa bean (5–40 wt/wt.% content of UFC). Si-PLS model was used to quantify the addition of UFC in FC. SVM model accurately discriminated the cocoa bean samples used. After cross-validation, the optimal identification rate was 100% in both the training set and prediction set at three principal components. For quantitative analysis, Si-PLS model was evaluated according to root mean square error of prediction (RMSEP) and coefficient of correlation in prediction (R_pred). The results revealed that Si-PLS model in this work was promising. The optimal performance of Si-PLS model showed an excellent predictive potential, RMSEP = 1.68 and R_pred = 0.98 in the prediction set. The overall results indicated that FT-NIR spectroscopy together with an appropriate multivariate algorithm could be employed for rapid identification of fermented and unfermented cocoa beans as well as the quantification of UFC down to 5% in FC for quality control management.     

Phenolic content and antioxidant capacity of hybrid variety cocoa beans

Cocoa (Theobroma cacao L.) is a major, economically important, international crop and has been associated with several nutritional benefits including high antioxidant capacity. New cocoa hybrids have been developed in Ghana that exhibit resistance to pest damage during storage. The aim of this work was to assess the phenolic content and antioxidant capacity of these new hybrids in comparison to more traditional cocoa varieties. Total extractable phenolics were similar in all the four hybrids tested ranging from 69.9 to 81.6 FAE g⁻¹. These levels were very similar to that extracted from traditional beans (73.8 ± 2.5 FAE g⁻¹). The “phenolic profile” was determined by HPLC. A total of 25 peaks was observed but there were only minor differences in this profile between traditional and hybrid bean extracts. Antioxidant capacity was determined using the FRAP assay and traditional beans were found to possess 12.4 μmol TE g⁻¹. In comparison the hybrid beans had antioxidant capacities ranging from 21.6 to 45.5 μmol TE g⁻¹, and these were significantly higher than in the traditional beans for three out of the four hybrids. Since the phenolic and antioxidant levels and in these hybrid varieties were either similar to, or higher than, that obtained from traditional beans, the introduction of these new varieties would be unlikely to impact detrimentally on these nutritional components of the beans.     

Analysis and occurrence of deoxynivalenol (DON) in cocoa

The primary objective of this study was to establish a current situation assessment of the possible occurrence of deoxynivalenol in cocoa and cocoa products. Since there was no analytic method for determining DON in cocoa and cocoa products, a special method was developed. The applicability and consistency of the method was confirmed by performing recovery assays on various cocoa products. A special post-column derivatisation procedure was used to increase selectivity and raise sensitivity by a factor of 80. The method’s limit of detection (LOD) was thereby reduced to 7 μg/kg; the limit of quantification (LOQ) was 14 μg/kg. The method was used to test 230 samples for possible DON content, ranging from cocoa beans to cocoa bean shells, nibs, cocoa liquor and cocoa powders through to finished cocoa-based products. In the case of cocoa beans and cocoa bean shells, DON content close to the detection limit was only determined in isolated cases. No DON content was detected in nibs, cocoa liquor, cocoa powders and finished cocoa-based products. Analogous to ochratoxin A and aflatoxins, the results show DON is more likely to be found in cocoa bean shells. Separation of shells during cocoa processing can reduce potential DON contents. Since no DON was determined in the fractions relevant for chocolate production, these assays show it does not represent a considerable issue for the cocoa and chocolate industry.     

Flavor formation and character in cocoa and chocolate: a critical review

Chocolate characters not only originate in flavor precursors present in cocoa beans, but are generated during post-harvest treatments and transformed into desirable odor notes in the manufacturing processes. Complex biochemical modifications of bean constituents are further altered by thermal reactions in roasting and conching and in alkalization. However, the extent to which the inherent bean constituents from the cocoa genotype, environmental factors, post-harvest treatment, and processing technologies influence chocolate flavor formation and relationships with final flavor quality, has not been clear. With increasing speciality niche products in chocolate confectionery, greater understanding of factors contributing to the variations in flavor character would have significant commercial implications.     

Extraction and determination of methylpyrazines in cocoa beans using coupled steam distillation-microdistillator

The formation of methylpyrazines was determined in fermented cocoa beans (Ivory Coast), in laboratory and industrially roasted samples. The determination of these methylpyrazines was studied by coupled steam distillation-microdistillation as the extraction method and gas chromatography using capillary column and a thermionic detector. The monomethyl-; 2,3-dimethyl-; 2,5-dimethyl-; 2,6-dimethyl-; trimethyl- and the tetramethylpyrazine were detected in non-roasted cocoa beans. Their concentration increased rapidly in laboratory roasted cocoa beans and industrial samples, only the tetramethylpyrazine showed a maximum peak of concentration. The principal compound was the tetramethylpyrazine in fermented and roasted cocoa beans. The determination of the different methylpyrazines in cocoa beans would permit both the evaluation of cocoa mass quality and the control of the cocoa roasting process.     

Vitamin E and Oxidative Stability of Soya Bean Oil Prepared with Beans at Various Moisture Contents Roasted in a Microwave Oven

Whole soya beans ( Glycine max ) at various moistures (96, 382 and 519 g kg⁻¹) were roasted by exposure to microwaves at a frequency of 2450 MHz and the effects on the tocopherols of soya beans were studied in rela-tion to chemical changes in the oils. The amounts of α-, β-, γ- and δ-tocopherols in the soya beans before microwave treatments ranged from 62 to 187, 43 to 89, 673 to 757 and 542 to 593 mg kg⁻¹ oil, respectively. Increasing moisture contents by soaking prevented, not only the reduction of tocopherols but also, the oxidative deterioration of soya bean oils during microwave roasting. The amounts of tocopherols still remained >80% of the original level in soaked soya bean oils after 20 min of roasting, and microwave roasting after soaking caused no significant differences ( P> 0·05, with few exceptions) in the chemical changes of the oils in comparison with those before soaking. These results implied that microwave roasting after soaking would be effective in making full-fat soya flour with high vitamin E without a burnt odour and browning from raw beans.     

Effects of incubation and polyphenol oxidase enrichment on colour, fermentation index, procyanidins and astringency of unfermented and partly fermented cocoa beans

Incubation of unfermented and partly fermented cocoa beans in acetate buffer, pH 5.5, at 45 °C increased yellowness, total colour differences and fermentation index value of the cocoa bean powders and decreased cocoa procyanidins (monomers to pentamers), and their astringency. Fermentation index and (–)‐epicatechin content, equivalent to those of fully fermented beans, were reached by unfermented beans after 4–8‐h incubation, but not by partly fermented beans even after 16 h. During incubation of partly fermented cocoa beans enriched with polyphenol oxidase, yellowness and fermentation index value were increased, whilst (–)‐epicatechin was decreased. Tyrosinase had a less significant effect in yellow colour formation, but showed a significant reduction of (–)‐epicatechin and increase in fermentation index compared with crude cocoa polyphenol oxidase. However, both enzymes have similar effects on procyanidin degradation and astringent taste reduction. Incubation of cocoa beans for 16 h increased the cut test score of unfermented and partly fermented beans by 50 and 30%, respectively.     

Changes in differently processed soya bean (Glycine max.) and lima bean (Phaseolus lunatus) with particular reference to their chemical composition and their mineral and some inherent anti-nutritional constituents

The effects of 3 processing techniques: cooking, roasting and autoclaving on the proximate chemical composition, the mineral content and some inherent toxic factors of soya and lima beans were investigated. The processing techniques generally reduced the crude fibre levels and enhanced the extractable fat in the soya and lima beans. The coefficients of variability for crude fibre and ether extract due to the processing techniques of soya and lima beans were 20.9%, 16.0% and 22.3%, 38.1%, respectively. In parallel with decreased ash content in the cooked bean samples, there was a decrease in the K, Mg, Na and P levels relative to the raw bean and also relative to the other processing techniques. Mineral contents of the autoclaved bean samples were generally similar to those of the raw (unprocessed) samples. Under the processing conditions, roasting caused the highest reduction in thioglucoside content (59%) in soya bean while cooking caused the highest reduction in lima bean (78%). Trypsin inhibitor activity (TIA) ranged between 0.59 mg/g sample in the cooked soya bean and 11.6 mg/g sample in the raw bean while the corresponding values for lima bean ranged between 0.59 and 6.3 mg/g sample. Cooking and roasting caused over 90% reduction of TIA, while autoclaving caused 64-69% reduction in both bean samples. Under the assay conditions, haemagglutinating activity was not detected in the cooked and autoclaved soya and lima beans. The need to prevent both functional and nutritional damage to food proteins and other nutrients, resulting from excessive heating, was discussed.     

Ghanaian Cocoa Bean Fermentation Characterized by Spectroscopic and Chromatographic Methods and Chemometrics

Abstract: Export of cocoa beans is of great economic importance in Ghana and several other tropical countries. Raw cocoa has an astringent, unpleasant taste, and flavor, and has to be fermented, dried, and roasted to obtain the characteristic cocoa flavor and taste. In an attempt to obtain a deeper understanding of the changes in the cocoa beans during fermentation and investigate the possibility of future development of objective methods for assessing the degree of fermentation, a novel combination of methods including cut test, colorimetry, fluorescence spectroscopy, NIR spectroscopy, and GC-MS evaluated by chemometric methods was used to examine cocoa beans sampled at different durations of fermentation and samples representing fully fermented and dried beans from all cocoa growing regions of Ghana. Using colorimetry it was found that samples moved towards higher a* and b* values as fermentation progressed. Furthermore, the degree of fermentation could, in general, be well described by the spectroscopic methods used. In addition, it was possible to link analysis of volatile compounds with predictions of fermentation time. Fermented and dried cocoa beans from the Volta and the Western regions clustered separately in the score plots based on colorimetric, fluorescence, NIR, and GC-MS indicating regional differences in the composition of Ghanaian cocoa beans. The study demonstrates the potential of colorimetry and spectroscopic methods as valuable tools for determining the fermentation degree of cocoa beans. Using GC-MS it was possible to demonstrate the formation of several important aroma compounds such 2-phenylethyl acetate, propionic acid, and acetoin and the breakdown of others like diacetyl during fermentation. Practical Application: The present study demonstrates the potential of using colorimetry and spectroscopic methods as objective methods for determining cocoa bean quality along the processing chain. Development of objective methods for determining cocoa bean quality will be of great importance for quality insurance within the fields of cocoa processing and raw material control in chocolate producing companies.