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Among the emerging prebiotics, galactooligosaccharide (GOS) has a remarkable value with health-promoting properties confirmed by several studies. In addition, the application of ohmic heating has been gaining prominence in food processing, due to its various technological and nutritional benefits. This study focuses on the transformative potential of ohmic heating processing (OH, voltage values 30 and 60 V, frequencies 100, 300, and 500 Hz, respectively) in prebiotic chocolate milk beverage (3.0 %w/v galactooligosaccharide) processing. Chemical stability of GOS was assessed along all the ohmic conditions. In addition, microbiological analysis (predictive modeling), physical analysis (color and rheology), thermal load indicators assessment, bioactivity values, and volatile compound was performed. HPAEC-PAD analysis confirmed GOS stability and volatile compound evaluation supported OH's ability to preserve flavor-associated compounds. Besides, OH treatments demonstrated superior microbial reduction and decreased thermal load indicators as well as the assessment of the bioactivity. In conclusion, OH presented was able to preserve the GOS chemical stability on chocolate milk beverages processing with positive effects of the intrinsic quality parameters of the product.

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BACKGROUND: The high quality and unique flavor and aroma of bean-to-bar chocolates have resulted in an increase in the consumption of these products. Nevertheless, cocoa beans may present inorganic contaminants from environmental and anthropogenic sources which can contribute to contamination of the chocolates, despite the fewer processing steps and few ingredients used in bean-to-bar manufacturing process compared to the industrial one. Therefore, this study aimed to evaluate the content of trace elements (As, Cd, Co, Cu, Hg, Pb, Se) in bean-to-bar chocolates and traceable cocoa beans from Brazil and Ecuador. METHODS: Bean-to-bar chocolate samples were acquired in Brazil (n=65) and Ecuador (n=10), considering the main products available: white, milk, semisweet and dark chocolate. Cocoa samples from dedicated farms (n=23) were analyzed for trace elements and inorganic contaminants regulated by Brazil and European agencies. Samples were mineralized using acid digestion (nitric acid and hydrogen peroxide) in a closed microwave-assisted system. Quantification of trace elements was performed using Inductively coupled plasma mass spectroscopy (ICP-MS) and Inductively coupled plasma optical emission spectroscopy (ICP OES) in optimized conditions. The analytical control was performed with certified reference materials (ERM BD512 - Dark Chocolate, Tort-2 and Tort-3 - Lobster Hepatopancreas and SRM 1547 - Peach leaves) and recoveries ranged between 84% and 105% for all elements. RESULTS: The trace element levels in the bean-to-bar chocolates were (mg/kg): As (<0.022-0.023), Cd (<0.002-0.74), Cu (0.11-21.2), Co (<0.003-1.88), Hg (<0.010-<0.010), Pb (<0.007-0.22), and Se (<0.029-0.35). The exposure assessment from inorganic contaminants in chocolates revealed up to 93% of provisional tolerable monthly intake (PTMI) for Cd and 123% of tolerable upper intake level (UL) for Co for children. Inorganic contaminants were also analyzed in cocoa beans from dedicated farms and Cd and Pb levels were found above the thresholds established by Brazil health agency. CONCLUSION: The results observed for both bean-to-bar chocolates and raw materials (cocoa beans from dedicated farms) indicated a need for monitoring these trace elements.

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This study is a bibliometric analysis and literature review on the use of oleogels (OGs), hydrogels (HGs) and hybrid gels (HYGs) in chocolate, compounds and spreads with the aim of reducing the saturated fat in these products. The articles were selected by analyzing titles, keywords and abstracts in the Web of Science (WoS), Scopus and Google Scholar databases. Supplementary documents were obtained from government sources, including patent registrations. The theoretical and practical aspects were critically analyzed, highlighting the main points of agreement and disagreement between the authors. The results revealed a lack of regulations and official guidelines that widely allow the use of OGs, HGs and HYGs in chocolate confectionery products. The type and characteristics of raw materials affect the properties of products. Replacing cocoa butter (CB) with OGs, HGs or HYGs also affects texture, melting point and behavior, and nutritional aspects. These substitutions can result in products with better sensory acceptance and health benefits, such as reducing saturated fat and promoting cardiovascular health. However, it is important to find the ideal combination and proportions of components to obtain the desired properties in the final products.

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Este trabalho teve como objetivo o estudo dos possíveis substitutos da sacarose em chocolates. O trabalho foi dividido em três capítulos a fim de se realizar um estudo abrangente sobre o tema. No Capítulo 1, concluiu-se, através de uma revisão bibliográfica, que a qualidade da textura e a estabilidade do chocolate são significativamente influenciadas pela existência de cristais específicos, especialmente os provenientes do açúcar. Por essa razão, é crucial examinar cuidadosamente a substituição desse componente. A literatura estudada menciona diversos edulcorantes e espessantes empregados para substituir a sacarose em chocolates, como os edulcorantes nutritivos e não-nutritivos, assim como os espessantes mais aplicados. O melhor edulcorante e fibra, considerando-se as literaturas estudadas, em termos de custo, propriedades reológicas e mouthfeel, para a substituição da sacarose em chocolates, seria o maltitol e a polidextrose. No Capítulo 2, foram comparados alguns chocolates amargos e ao leite, com e sem sacarose presentes no mercado. Estes chocolates apresentaram comportamento calorimétrico indicando pico de fusão, pico de caramelização e pico de carbonização variáveis de acordo com a formulação. Os chocolates apresentaram forças de ruptura semelhantes, entretanto, o dimensional das amostras influenciou na textura dos produtos. O tipo de edulcorante e os ingredientes utilizados, influenciaram no fluxo de escoamento e nas características físicas, como a cor dos chocolates. No Capítulo 3, foram formulados dois chocolates amargos, um com adição de açúcar e um com a adição de uma mistura comercial (contendo fibra de mandioca/tapioca e edulcorantes), disponível comercialmente, porém sem estudos anteriores de aplicação em chocolates, em substituição à sacarose. As amostras foram comparadas, entre si e em relação aos valores encontrados na literatura, quanto as suas propriedades nutricionais, físico-químicas e reológicas. A substituição da sacarose, pela mistura comercial, demonstrou comportamento próximo das amostras de mercado estudadas no Capítulo 2. Concluiu-se que a mistura comercial cumpre as funções da sacarose em formulação de chocolate amargo diet, mesmo que alterando algumas das propriedades físico-químicas e reológicas. Entretanto, esta mistura não é indicada para uso em formulações de chocolates forneáveis, pois apresenta pico de carbonização em temperatura precoce

This work aimed to study possible sucrose substitutes in chocolates. The study was divided into three chapters to conduct a comprehensive examination of the topic. In Chapter 1, it was concluded, through a literature review, that the texture quality and stability of chocolate are significantly influenced by the presence of specific crystals, especially those derived from sugar. For this reason, it is crucial to carefully examine the replacement of this component. The literature studied mentions various sweeteners and thickeners used to replace sucrose in chocolates, including both nutritive and non-nutritive sweeteners, as well as commonly applied thickeners. According to the literature reviewed, the best sweetener and fiber for the replacement of sucrose in chocolates, considering cost, rheological properties, and mouthfeel, would be maltitol and polydextrose. In Chapter 2, some dark and milk chocolates without sucrose available in the market were compared. These chocolates exhibited calorimetric behavior indicating variable melting peaks, caramelization peaks, and carbonization peaks according to the formulation. The chocolates exhibited similar breaking strengths, however, the dimensional characteristics of the samples influenced the texture of the products. The type of sweetener and ingredients used influenced the flow behavior and physical characteristics, such as the color of the chocolates. In the Chapter 3, two dark chocolates were formulated, one with sugar added and one with the addition of a commercial blend (containing cassava/tapioca fiber and sweeteners), commercially available, but without previous studies of application in chocolates, as a substitute for sucrose. The samples were compared, both between themselves and in relation to the values found in the literature, regarding its nutritional, physicochemical, and rheological properties. The replacement of sucrose with the commercial blend demonstrated behavior similar to the market samples studied in Chapter 2. It is concluded that the commercial blend fulfills the functions of sucrose in diet dark chocolate formulation, even changing some of the physicochemical and rheological properties of the chocolate. However, this blend is not recommended for the use in bakeable chocolate formulations, as it exhibits an early peak of carbonization

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Aiming to improve the quality of cocoa, preconditioning of cocoa after harvesting and before fermentation has become an on-farm processing step of great interest in recent times. The present work aimed to evaluate the influence of a pre-drying process on the volatile composition of Ecuadorian bulk (Forastero and CCN-51) and fine-flavour (ETT103 and LR14) cocoa at the end of primary processing. A total of 63 volatile compounds including aldehydes, alcohols, acids, ketones, esters, terpenes, lactones and other miscellaneous compounds were identified in cocoa samples by HS-SPME-GC-MS. The use of a pre-drying step revealed a varietal homogenization and a reduction in the fermentation time, making this preconditioning step an inexpensive and attractive option for farmers. Moreover, different varietal behaviour was observed after pre-drying, the fine-flavour varieties obtaining a clear improvement in aromatic quality with higher levels of compounds imparting positive notes.

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The chemical composition of dark chocolate has a significant impact on its complex flavor profile. This study aims to investigate the relationship between the volatile chemical composition and perceived flavor of 54 dark chocolate samples made from Trinitario cocoa beans from the Dominican Republic. The samples were evaluated by a trained panel and analyzed using gas chromatography-mass spectrometry (GC-MS) to identify and quantify the volatile compounds. Predictive models based on a partial least squares regression (PLS) allowed the identification of key compounds for predicting individual sensory attributes. The models were most successful in classifying samples based on the intensity of bitterness and astringency, even though these attributes are mostly linked to non-volatile compounds. Acetaldehyde, dimethyl sulfide, and 2,3-butanediol were found to be key predictors for various sensory attributes, while propylene glycol diacetate was identified as a possible marker for red fruit aroma. The study highlights the potential of using volatile compounds to accurately predict chocolate flavor potential.

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Rapid assessment of pesticide residues ensures cocoa bean quality and marketability. In this study, a portable FTIR instrument equipped with a triple reflection attenuated total reflectance (ATR) accessory was used to screen cocoa beans for pesticide residues. Cocoa beans (n = 75) were obtained from major cocoa growing regions of Peru and were quantified for pesticides by gas chromatography (GC) or liquid chromatography (LC) coupled with mass spectrometry (MS). The FTIR spectra were used to detect the presence of pesticides in cocoa beans or lipid fraction (butter) by using a pattern recognition (Soft Independent Modeling by Class Analogy, SIMCA) algorithm, which produced a significant discrimination for cocoa nibs (free or with pesticides). The variables related to the class grouping were assigned to the aliphatic (3200-2800 cm-1) region with an interclass distance (ICD) of 3.3. Subsequently, the concentration of pesticides in cocoa beans was predicted using a partial least squares regression analysis (PLSR), using an internal validation of the PLRS model, the cross-validation correlation coefficient (Rval = 0.954) and the cross-validation standard error (SECV = 14.9 mg/kg) were obtained. Additionally, an external validation was performed, obtaining the prediction correlation coefficient (Rpre = 0.940) and the standard error of prediction (SEP = 16.0 µg/kg) with high statistical performances, which demonstrates the excellent predictability of the PLSR model in a similar real application. The developed FTIR method presented limits of detection and quantification (LOD = 9.8 µg/kg; LOQ = 23.1 µg/kg) with four optimum factors (PC). Mid-infrared spectroscopy (MIR) offered a viable alternative for field screening of cocoa.

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Cocoa currently faces differentiation processes toward niches of specialty products, leading to greater competitiveness for producers who must compete with products differentiated by their integral quality regarding their organoleptic characteristics, such as fine-flavor cocoa and their functional characteristics. Quality is influenced by the genetic variety of the cultivars on the one hand, and the correct postharvest processing operations of cocoa seeds, on the other. During the transformation operations, the native chemical compounds of the seeds, especially proteins, carbohydrates, and polyphenols, are transformed and generate other compounds called flavor precursors, which are responsible for defining the product quality. In this sense, the analysis of the most relevant chemical compounds in cocoa is essential to guarantee higher overall quality. Similarly, understanding the fundamental aspects that affect fine-flavor cocoa production is crucial for improving transformation processes. Therefore, reliable and robust analytical techniques are required to detect and quantify these chemical compounds. This review highlights the main techniques used to analyze essential cocoa metabolites and derived products throughout all postharvest transformation stages: from cocoa seeds to chocolate bar, offering an overview of the sample preparation methods and the analytical and imaging methodologies often employed to characterize qualifying cocoa products.

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A non-segregated kinetic model is proposed to describe a fermentation process of agro-industrial residues derived via cocoa (mucilage juice) by Pichia kudriavzevii. The novel proposed hybrid model is based on a multiple coupling reaction mechanisms (structured) to describe the kinetics of substrate consumption, biomass, carbon dioxide, and ethanol, coupled to an unstructured model for the activity enzyme. The parameters of the kinetic model are estimated by non-linear least-squares curve fitting using the Marquardt-Levenberg algorithm. In addition, numerical simulations were compared with the experimental data via residual graphs. The effectiveness of the model was statistically evaluated using dimensionless efficiency coefficients under different initial conditions. A global sensitivity analysis was applied (Fisher's information matrix). The experimental results of the batch reactor showed a maximum ethanol concentration of 29 g/L, with a yield of 0.48 g-ethanol/g-glucose and a productivity of 0.30 g/L h. The method determined that the cell formation coefficient and the specific substrate consumption rate (θ1 and θ2) directly influence most of the states of our system. The proposed scheme is particularly suitable to assist in the rational design of cell factory properties or fermentation processes because it can represent the complex biochemistry in more detail and under different initial experimental conditions; the above reveals that the generated model is robust and can be considered for control and optimization purposes.

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This study aimed to evaluate the volatile compounds of chocolates made of Brazilian cocoas and statistically track them according to the products' sensorial profile in order to relate them to consumers' acceptance by preference map methodology. The intensity of the chocolate, acidity, woody, smoked, green, floral, burned, musty, and cocoa notes from chocolates produced with cocoa from different Brazilian states were analyzed by a trained panel and by 128 consumers. Samples from Côte d'Ivoire, which is known for its high-quality chocolate, were evaluated for comparison. Solid-phase microextraction headspace sampling/gas chromatography-mass spectrometry was employed to evaluate the samples' volatile compounds. One hundred volatile compounds were identified within the samples. The results from the preference maps showed that the maximum preference was found for chocolate made of cocoa from Rondônia, Bahia, and Espírito Santo and Côte d'Ivoire and organic samples from Pará. The ideal sample point was characterized by intense chocolate, floral, and woody notes and mild green and burned notes. The presence of furfural, 3-methyl butanal, phenethyl acetate, 2-phenyl-5-methyl-2-hexenal, methyl pyrazine, phenethyl acetate, 2-phenyl-5-methyl-2-hexenal, and tetramethyl pyrazine were shown to be important for consumer acceptance in the ideal product, whereas the presence of (Z)-2-heptenal and 2-pentyl furan may increase consumer rejection. 2,3-Methyl pyrazine, methyl pyrazine, and 2,3-butanediol, which are important volatile compounds previously reported in the literature, were statistically tracked to both positive and negative sample attributes and must be better explored concerning consumers' acceptance of chocolates.

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The processing of cupuassu (Theobroma grandiflorum Schum) beans after fermentation gives a chocolate-like product, the cupulate. The high amount of pulp adhered to the seeds hinders the fermentation. Consequently, it is necessary to depulp the seeds to perform the process, even though the pulp contains important substrates for the formation of flavor precursors. To verify whether the complete or partial removal of the pulp influences the sensory characteristics of the product, fermentation was performed with three pulp concentrations (0, 7.5, and 15%) and two schemes of turning for aeration of the mass: fixed (R1) and according to the temperature (R2), in a total of six experiments (0R1, 0R2, 7.5R1, 7.5R2, 15R1 and 15R2). The beans were processed to obtain cupulates, which were submitted to tests performed with consumers, to express their preference and attributes (acceptance, purchase intent, Check All That Apply - CATA), and then to tests with a trained panel, the Quantitative Descriptive Profile (QDP) to characterize the samples. Both tests showed the consumers' perceptions that the cupulates have peculiar sensory characteristics. In the Consumer Test, through the Preference Mapping, all the samples of cupulates obtained from R1 conditions were preferred. In the penalty analysis, these same samples showed positive attributes that mask the negative attributes. Both CATA and QDP results showed that cupulate samples produced from seeds with a higher amount of pulp (15R1 and 15R2) had a higher number of positive mentions, for their fruity and floral flavors. The research also demonstrated that all samples gave the perception of an earthy taste, an important reason for consumer rejection, as well as a bad residual flavor. Thus, the results showed that the presence of the pulp in the fermentation environmental is important to the formation of flavor compounds and improving the sensory acceptance of the products.

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Regulation of maximum levels of cadmium in chocolate is an issue for cacao exportation from many parts of Latin America, including Colombia. These limits are related to the final product, but buyers often request maximum levels of Cd in the beans. However, to date, there is neither a clear understanding of the relationship between the specified levels of Cd in chocolate and cocoa derivatives and levels in harvested beans or soil nor of the effect of post-harvest processes on the levels of Cd in the final product. To address this, the fate of Cd concentration from soil to chocolate bar was followed in a single farm in Santander district, Colombia. The concentration of Cd in soils was measured using ICP-OES and correlated with soil pH, soil organic matter (SOM), and the use of P-based fertilisers. Cd concentrations were also measured in unfermented seeds, fermented and dried beans, shell, nibs, and chocolate. SOM (2.93-3.78%), soil pH (4.7-4.9), soil P concentration (120-132 mg kg-1) affect Cd availability. However, it is still unclear whether Cd concentration of P-based fertilisers (3-30 mg kg-1) is important or not. While post-harvest treatments did not affect the Cd concentration of beans (4.17 ± 0.8 mg kg-1 on average), the removal of the shell (6.57 mg kg-1) from the nibs (3.28 mg kg-1), as well as the percentage of cocoa mass used contributes to a reduction in Cd concentration in the chocolate bar (1.60 mg kg-1). This study provides clear indications on where research into mitigation measures should be focussed, as well as indicating the importance of carrying out analyses for Cd in the nib or cacao mass, rather than the whole bean, reducing Cd concentration by up to 40%.

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Yeasts are one of the main ingredients responsible for flavor precursors production associated with sensorial characteristics in chocolate. Using wild yeast isolated from cocoa beans fermentation is emerging as a strategy for developing starter cultures. However, the volatile compounds (VCs) produced by yeasts are not yet known. This study aimed to select wild yeasts with the potential to produce volatile compounds associated with desirable flavor attributes. A total of 150 wild yeasts strains were isolated from the spontaneous cocoa beans fermentation, of which 40 were identified by morphology and physiological features. VCs produced were identified and quantified using SPME-GC-MS and GC-FID and profiles were evaluated statistically by PCA and cluster analysis for the compounds that had a high odor threshold value. Thirty-six VCs produced by these yeasts were identified into six main families, namely esters, alcohols, acids, aldehydes, ketones, and pyrazines. PCA showed the separation of the yeasts into two main clusters. Strains, Y195 and Y246, belong to the first cluster and are the highest producers of alcohols related to floral perceptions. In the second cluster, thirty-three yeasts were grouped by their ability to produce esters. Of all of them, Y110MRS stood out for producing 2-phenyl ethyl acetate and isoamyl acetate associated with fruity perceptions. This screening allowed us to identify yeasts that produced VCs of technological interest and which could be used to develop a starter culture.

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Metagenomic studies about cocoa fermentation have mainly reported on the analysis of short reads for determination of operational taxonomic units. However, it is also important to determine metagenome-assembled genomes (MAGs), which are genomes deriving from the assembly of metagenomics. For this research, all the cocoa metagenomes from public databases were downloaded, resulting in five data sets: one from Ghana and four from Brazil. In addition, in silico approaches were used to describe putative phenotypes and the metabolic potential of MAGs. A total of 17 high-quality MAGs were recovered from these microbiomes, as follows: (i) for fungi, Yamadazyma tenuis (n = 1); (ii) lactic acid bacteria, Limosilactobacillus fermentum (n = 5), Liquorilactobacillus cacaonum (n = 1), Liquorilactobacillus nagelli (n = 1), Leuconostoc pseudomesenteroides (n = 1), and Lactiplantibacillus plantarum subsp. plantarum (n = 1); (iii) acetic acid bacteria, Acetobacter senegalensis (n = 2) and Kozakia baliensis (n = 1); and (iv) Bacillus subtilis (n = 1), Brevundimonas sp. (n = 2), and Pseudomonas sp. (n = 1). Medium-quality MAGs were also recovered from cocoa microbiomes, including some that, to our knowledge, were not previously detected in this environment (Liquorilactobacillus vini, Komagataeibacter saccharivorans, and Komagataeibacter maltaceti) and others previously described (Fructobacillus pseudoficulneus and Acetobacter pasteurianus). Taken together, the MAGs were useful for providing an additional description of the microbiome of cocoa fermentation, revealing previously overlooked microorganisms, with prediction of key phenotypes and biochemical pathways. IMPORTANCE The production of chocolate starts with the harvesting of cocoa fruits and the spontaneous fermentation of the seeds in a microbial succession that depends on yeasts, lactic acid bacteria, and acetic acid bacteria in order to eliminate bitter and astringent compounds present in the raw material, which will be further roasted and grinded to originate the cocoa powder that will enter the food processing industry. The microbiota of cocoa fermentation is not completely known, and yet it advanced from culture-based studies to the advent of next-generation DNA sequencing, with the generation of a myriad of data that need bioinformatic approaches to be properly analyzed. Although the majority of metagenomic studies have been based on short reads (operational taxonomic units), it is also important to analyze entire genomes to determine more precisely possible ecological roles of different species. Metagenome-assembled genomes (MAGs) are very useful for this purpose; here, MAGs from cocoa fermentation microbiomes are described, and the possible implications of their phenotypic and metabolic potentials are discussed.

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High-yield resistant hybrids are used in cocoa fermentation and result in chocolates with different sensorial profiles. This work aimed to characterize the fermentation microbiologically and physicochemically. Hybrids CEPEC 2004, FA13, PH15, and CEPEC 2002 were used for fermentation. The yeast, acetic acid bacteria, lactic acid bacteria, and mesophilic bacteria population were evaluated in their respective medium. Carbohydrates and acids were detected using a high-performance liquid chromatography system, and volatiles were analyzed using gas chromatography-mass spectrometry equipment. Finally, a consumer acceptance test followed by a check-all-that-apply question and a temporal dominance of sensations assessment was performed in chocolate. The fermentation resulted in a typical succession: yeast-dominated at first, followed by lactic acid, acetic acid, and mesophilic bacteria. In the pulp, carbohydrates and citric acid were consumed. Low concentrations of acetic acid (0.09-1.75 g/kg) were detected. Acids, esters, and alcohols were the most abundant groups. The chocolate profile resulted in sweet, acidic, and fruity, satisfying consumers' tastes. PRACTICAL APPLICATION:  The cocoa hybrid-mix fermentation can improve the fermentation process and chocolate quality.  The mixture generated a different sensory profile in comparison to other fermentations.  The fruity chocolate was accepted and liked by consumers.

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Se evaluó la composición proximal, el contenido de polifenoles totales y la capacidad antioxidante de un chocolate oscuro formulado a partir de granos de cacao orgánico, seco, y no sometido al proceso fermentativo. Este producto fue elaborado con insumos y procesos desarrollados en la Granja Experimental El Cairo, ubicada en la Universidad Nacional de Colombia (GEL-UN), departamento de Arauca, Colombia. El contenido de grasa se encontró entre los rangos conocidos (>50%), siendo una característica ligada al origen genético, y a las condiciones climáticas. El alto contenido de proteínas (16,21%), evidencia que se trata de granos de cacao secos no fermentados. Tal contenido juega un papel importante en la formación de los precursores del sabor y aroma, en chocolates finos y exquisitos. El pH determinado es un indicativo de un chocolate amargo, con bajo índice de fermentación y de baja acidez acética o láctica. El contenido de polifenoles totales se relaciona de manera directa con la alta actividad antioxidante de este chocolate. Se concluye que el tipo de material genético usado como insumo, así como los procesos implementados para la obtención del chocolate, en GEL-UN, contribuyeron para la obtención de un producto tipo comercial, con propiedades funcionales, entre ellas las relacionadas con la capacidad antioxidante(AU)

Thee proximal chemical composition, the content of total polyphenols, and the antioxidant capacity of a dark chocolate formulated from organic, dry cocoa beans and not subjected to the fermentation process were evaluated. This product was made with inputs and processes developed at the El Cairo Experimental Farm, located at the Universidad Nacional de Colombia (CEF-UN), Arauca, Colombia. The fat content was found between the known ranges (> 50%), being a characteristic linked to the genetic origin, and to the climatic conditions. The high protein content (16.21%) evidenced that it´s unfermented dry cocoa beans. Such content plays an important role in the formation of precursors of flavor and aroma, in fine and exquisite chocolates. The determined pH is indicative of a dark chocolate, with a low fermentation index and low acetic or lactic acidity. The content of total polyphenols is directly related to the high antioxidant activity of this chocolate. It is concluded that the type of genetic material used as input, as well as the processes implemented to obtain chocolate, in CEF-UN, contributed to obtaining a commercial type product, with functional properties, including those related to antioxidant capacity(AU)

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The Near (NIR) and Mid (MIR) Infrared Spectroscopy associated with chemometric techniques were used to determine the cocoa solids content in chocolates and detect possible adulterations. Five chocolate formulations (30% to 90%) were produced with different cocoa solids concentrations and 110 commercial samples from 10 different countries with varying concentrations of cocoa solids (30% to 88%) were acquired. All repetions of the produced and commercial chocolates were evaluated using NIR and MIR. Spectroscopic data were submitted to multivariate techniques of Principal Component Analysis (PCA) and Partial Least Squares Regression (PLS). For both spectroscopy techniques, the PCA of the 5 formulations formed 5 distinct groups regarding the cocoa solids and the commercial samples showed a behavior pattern similar to the produced samples. For PLS, the regression equations showed high predictive capacity, with correlation coefficients above 90 and RMSECV values of 0.70 and 1.22, for NIR and MIR, respectively. These models highlighted, approximately, 14% of the commercial samples as possible adulterated products.

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Macro- and microelement determination in chocolate bars by microwave-induced plasma optical emission spectrometry (MIP OES) was evaluated after microwave-assisted sample digestion. Optimization of the sample digestion was carried out, and the recommended conditions were obtained at a temperature of 190 °C, with a digestion time of 40 min and in a mixture constituted by 2.3 mL of nitric acid, 1.0 mL of hydrogen peroxide and 4.7 mL of water. The method was applied in the analysis of chocolate bars, and the concentration ranges of the elements determined were (in mg kg-1): Ca (653-3096); Cr (<0.6-2.8); Cu (<0.16-19.5); Fe (<1.6-227); Mg (147-2775); K (3554-8573); Mn (<0.03-25.2); Na (45.6-1095); Ni (3.2-10.2); P (1111-22594) and Zn (4.8-33.3). The association of the proposed microwave-assisted acid digestion with the MIP OES technique was adequate for multi-element determination in chocolate bars for routine analysis.

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Chocolate is an important source of free bioactive amines and amino acids which play important roles in human health. Considering the limited information on the bioaccessibility of these compounds from chocolate, the objective of this study was to characterize their profiles and bioaccessibility in 70% cocoa dark chocolate through in vitro simulation of oral, gastric and intestinal digestions. Seven amines were detected; polyamines were predominant before in vitro digestion, whereas tyramine, cadaverine and spermidine after digestion. All amines showed high bioaccessibility with slight influence of digestive enzymes. Amines increased after gastrointestinal digestion: tyramine (13-fold), tryptamine (9-fold), others (2.4-4.2-fold) and histamine appeared. All amino acids, GABA and ammonia were detected in chocolate, and their contents increased after in vitro digestion due to digestive enzymes (4.6, 2.8 and 2.1, respectively). Dark chocolate protein is a good source of tryptophan, phenylalanine + tyrosine, isoleucine, histidine, but limiting for lysine, leucine, and threonine.

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This study aimed to identify the volatile compounds in the fermented and dried cocoa beans conducted with three distinct inoculants of yeast species due to their high fermentative capacity: Saccharomyces cerevisiae, Pichia kudriavzevii, the mixture in equal proportions 1:1 of both species, and a control fermentation (with no inoculum application). Three starter cultures of yeasts, previously isolated and identified in cocoa fermentation in the municipality of Tomé-Açu, Pará state, Brazil. The seeds with pulp were removed manually and placed in wooden boxes for the fermentation process that lasted from 6 to 7 days. On the last day of fermentation, the almonds were packaged properly and placed to dry (36 °C), followed by preparation for the analysis of volatile compounds by GC-MS technique. In addition to the control fermentation, a high capacity for the formation of desirable compounds in chocolate by the inoculants with P. kudriavzevii was observed, which was confirmed through multivariate analyses, classifying these almonds with the highest content of aldehydes, esters, ketones and alcohols and low concentration of off-flavours. We conclude that the addition of mixed culture starter can be an excellent alternative for cocoa producers, suggesting obtaining cocoa beans with desirable characteristics for chocolate production, as well as creating a product identity for the producing region.

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