Microwave-assisted unfermented cocoa bean: improving flavor precursor after acetic acid submersion.

Objective was to enhance flavor precursors of unfermented cocoa beans by soaking beans in acetic acid and further heated by microwave. Acetic acid concentration, microwave power, and microwave exposure time were investigated and screened using a one-variable-at-a-time approach. Optimum condition for degree of hydrolysis (DH) was determined by Response Surface Methodology using Box-Behnken Design. Results showed that flavor precursors increased at a higher acetic acid concentration, microwave power, and microwave exposure time. Optimum condition was achieved at acetic acid concentration of 1.21 M, microwave power at 450 W, and microwave exposure time of 4 min. The microwave-assisted cocoa bean had a DH of 38.99% and a reducing sugar of 0.98%. Microwave-assisted heating increased amino acid content, especially hydrophobic amino acids as flavor precursors, and the main volatile compounds, especially aldehyde and pyrazine. Thus, microwave-assisted heating is a promising alternative to improve flavor precursors of unfermented cocoa beans.

Classification of Organic and Conventional Cocoa Beans Using Laser-Induced Fluorescence Spectroscopy Combined with Chemometric Techniques.

The craving for organic cocoa beans has resulted in fraudulent practices such as mislabeling, adulteration, all known as food fraud, prompting the international cocoa market to call for the authenticity of organic cocoa beans before export. In this study, we proposed robust models using laser-induced fluorescence (LIF) and chemometric techniques for rapid classification of cocoa beans as either organic or conventional. The LIF measurements were conducted on cocoa beans harvested from organic and conventional farms. From the results, conventional cocoa beans exhibited a higher fluorescence intensity compared to organic ones. In addition, a general peak wavelength shift was observed when the cocoa beans were excited using a 445 nm laser source. These results highlight distinct characteristics that can be used to differentiate between organic and conventional cocoa beans. Identical compounds were found in the fluorescence spectra of both the organic and conventional ones. With preprocessed fluorescence spectra data and utilizing principal component analysis, classification models such as Linear Discriminant Analysis (LDA), Support Vector Machine (SVM), Neural Network (NN) and Random Forest (RF) models were employed. LDA and NN models yielded 100.0% classification accuracy for both training and validation sets, while 99.0% classification accuracy was achieved in the training and validation sets using SVM and RF models. The results demonstrate that employing a combination of LIF and either LDA or NN can be a reliable and efficient technique to classify authentic cocoa beans as either organic or conventional. This technique can play a vital role in maintaining integrity and preventing fraudulent practices in the cocoa bean supply chain.

An in-depth multiphasic analysis of the chocolate production chain, from bean to bar, demonstrates the superiority of Saccharomyces cerevisiae over Hanseniaspora opuntiae as functional starter culture during cocoa fermentation.

Hanseniaspora opuntiae is a commonly found yeast species in naturally fermenting cocoa pulp-bean mass, which needed in-depth investigation. The present study aimed at examining effects of the cocoa isolate H. opuntiae IMDO 040108 as part of three different starter culture mixtures compared with spontaneous fermentation, regarding microbial community, substrate consumption, and metabolite production dynamics, including volatile organic compound (VOC) and phytochemical compositions, as well as compositions of the cocoa beans after fermentation, cocoa liquors, and chocolates. The inoculated H. opuntiae strain was unable to prevail over background yeasts present in the fermenting cocoa pulp-bean mass. It led to under-fermented cocoa beans after four days of fermentation, which was however reflected in higher levels of polyphenols. Cocoa fermentation processes inoculated with a Saccharomyces cerevisiae strain enhanced flavour production during the fermentation and drying steps, which was reflected in richer and more reproducible aroma profiles of the cocoa liquors and chocolates. Sensory analysis of the cocoa liquors and chocolates further demonstrated that S. cerevisiae led to more acidic notes compared to spontaneous fermentation, as a result of an advanced fermentation degree. Finally, different VOC profiles were found in the cocoa beans throughout the whole chocolate production chain, depending on the fermentation process.

Untargeted Metabolomics Analysis for Studying Differences in High-Quality Colombian Cocoa Beans.

Colombia is a producer of fine cocoa, according to the International Cocoa Organization; however, most of its exports are in the ordinary cocoa category. To remedy this situation, several national organizations are working to create technological platforms for small producers to certify the quality of their beans. The objective of this study was to identify differential chemical markers in 36 cocoa bean samples from five Colombian departments and associate them with cocoa quality properties. For this purpose, a non-targeted metabolomics approach was performed using UHPLC-HRMS, along with sensory and physicochemical analyses. The 36 samples did not differ in sensory quality, polyphenol content, and theobromine/caffeine ratio. However, the multivariate statistical analysis allowed us to differentiate the samples into four clusters. In addition, a similar grouping of the samples was also observed in the physical analyses. The metabolites responsible for such clustering were investigated with univariate statistical analysis and presumptively identified by comparison of experimental mass spectra with those reported in databases. Alkaloids, flavonoids, terpenoids, peptides, quinolines, and sulfur compounds were identified as discriminants between sample groups. Here, it was presented the metabolic profiles as an important chemical feature for further studies in quality control and more specific characterization of fine cocoa.

Screening Wild Yeast Isolated from Cocoa Bean Fermentation Using Volatile Compounds Profile.

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.

Cocoa By-Products: Characterization of Bioactive Compounds and Beneficial Health Effects.

The annual production of cocoa is approximately 4.7 million tons of cocoa beans, of which only 10% corresponds to the cocoa bean and the remaining value corresponds to a high number of residues, cocoa bean shell, pulp and husk. These by-products are a source of nutrients and compounds of notable interest in the food industry as possible ingredients, or even additives. The assessment of such by-products is relevant to the circular economy at both environmental and economic levels. Investigations carried out with these by-products have shown that cocoa husk can be used for the production of useful chemicals such as ketones, carboxylic acids, aldehydes, furans, heterocyclic aromatics, alkylbenzenes, phenols and benzenediols, as well as being efficient for the removal of lead from acidic solutions, without decay in the process due to the other metals in this matrix. The fibre present in the cocoa bean shell has a considerable capacity to adsorb a large amount of oil and cholesterol, thus reducing its bioavailability during the digestion process, as well as preventing lipid oxidation in meats, with better results compared to synthetic antioxidants (butylated hydroxytoluene and ß-tocopherol). Finally, cocoa pulp can be used to generate a sweet and sour juice with a natural flavour. Thus, this review aimed to compile information on these by-products, focusing mainly on their chemical and nutritional composition, simultaneously, the various uses proposed in the literature based on a bibliographic review of articles, books and theses published between 2000 and 2021, using databases such as Scopus, Web of Science, ScieLO, PubMed and ResearchGate.

Volatile organic compounds and sensory profile of dark chocolates made with cocoa beans fermented with Pichia kudriavzevii and Hanseniaspora thailandica.

Volatile organic compounds (VOCs) are important to determine the aroma and sensory perception of cocoa. Starter cultures can modulate the volatile profile of cocoa beans during fermentation. This study aimed to determine the VOCs and sensory of chocolates produced using cocoa beans fermented with yeast starters (Pichia kudriavzevii (MH979681), Hanseniaspora thailandica (MH979675) and the mixture of the two yeasts (Mix)). The VOCs of chocolates were determined by Head-Space Solid Phase Microextraction followed by Gas Chromatography-Mass Spectrophotometry. Sensory analysis was determined by using trained panels. VOCs profiles of chocolates produced using beans fermented with HT, PK or Mix were noticeably different from Ghana and control chocolates (no starter). The addition of yeast starters during cocoa fermentation produced chocolates that were preferred by trained panels. Bitterness and astringency were the more intense flavour attributes in chocolates produced using cocoa beans added with yeast starters. The chocolate produced using cocoa beans fermented with PK was the most acidic; whereas chocolate produced using beans fermented with Mix had the sweetest taste. The addition of PK or HT is helpful in producing chocolate with a distinct flavour.

Effect of spontaneous fermentation location on the fingerprint of volatile compound precursors of cocoa and the sensory perceptions of the end-chocolate.

Cocoa pod-opening delay and bean fermentation promote the organoleptic quality of chocolate. The present research investigated the changes in the volatile fingerprint of cocoa harvested at a traditional plantation. Cocoa beans extracted from 2-days pod-opening delay were simultaneously fermented for 5 days using container and then sun-dried to 7-8% moisture content at five different locations: Akoupé, San Pedro, Soubré, Djekanou and Daloa. The aromatic analysis were done on cocoa using the HS-SPME-GC/MS technique. Professional panelists evaluated the sensory perceptions of the chocolate. The results shows that cocoa fermented in both Daloa and Soubré regions were differentiated by 2,3-butanediol while those processed in other regions presented highest acetoin content. However, fermented cocoa from Soubré region exhibited most amount of 2,3-butanediol, diacetate A whereas 2,3,5,6-tetramethylpyrazine differentiated those from Daloa region. Sensory properties of chocolate were not linked to the aromatic compound precursors profile of beans. The fermentation performed in San Pédro region promote both the generation of more desirable aromatic compounds of cocoa and sensory attributes of the finished chocolate. The fermentation location generates a greater differentiation of the volatile fingerprint of cocoa and the sensory perceptions of the finished chocolate.

Methodology development based on "dilute and shoot" and QuEChERS for determination of multiple mycotoxins in cocoa by LC-MS/MS.

This work proposes an extraction method based on the "dilute and shoot" approach and QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) for the simultaneous determination of 42 mycotoxins (34 quantified and 8 qualitatively studied) in dried cocoa bean samples. The purpose of the developed methodology was the reduction of co-extractives from the matrix and an efficient extraction without a cleanup step, and subsequent analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). In order to obtain the best extraction conditions, gravimetric tests were performed and parameters that influenced the extraction efficiency were evaluated, such as the proportion of extraction phases, amount of salt, acidification, and extraction time. The performance of the developed method was evaluated to ensure its reliability. Considering the recovery range of 70-120% as an accuracy parameter, four of the mycotoxins under study (acetyl T-2, tenuazonic acid, wortmannin, and zearalenone) showed undesirable values at one of the levels evaluated. The repeatability of the method was assessed for 34 mycotoxins by the relative standard deviation (RSD%) of the responses, and all presented satisfactory values. The quantification limits ranged from 1.0 to 33.0 µg kg-1. Modification of the extraction methods made it possible to simultaneously analyze multiple mycotoxins, eliminating the need for the cleanup step, which led to analyte losses. The proposed methodology has a low cost, which makes it advantageous in routine analysis. It also has the potential for scope extension to cocoa-based foods, which are naturally exposed to a greater variety of mycotoxins. Graphical abstract.

Cocoa beans improve mitochondrial biogenesis via PPARγ/PGC1α dependent signalling pathway in MPP+ intoxicated human neuroblastoma cells (SH-SY5Y).

Polyphenols are shown to protect from or delay the progression of chronic neurodegenerative diseases. Mitochondrial dysfunction plays a key role in the pathogenesis of Parkinson's disease (PD). This study was aims to gain insight into the role of ahydroalcoholic extract of cocoa (standardised for epicatechin content) on mitochondrial biogenesis in MPP+ intoxicated human neuroblastoma cells (SHSY5Y). The effects of cocoa on PPARγ, PGC1α, Nrf2 and TFAM protein expression and mitochondrial membrane potential were evaluated. A pre-exposure to cocoa extract decreased reactive oxygen species formation and restored mitochondrial membrane potential. The cocoa extract was found to up-regulate the expression of PPARγ and the downstream signalling proteins PGC1α, Nrf2 and TFAM. It increased the expression of the anti-apoptotic protein BCl2 and increased superoxide dismutase activity. Further, the cocoa extract down-regulated the expression of mitochondria fission 1 (Fis1) and up-regulated the expression of mitochondria fusion 2 (Mfn2) proteins, suggesting an improvement in mitochondrial functions in MPP+ intoxicated cells upon treatment with cocoa. Interestingly, cocoa up-regulates the expression of tyrosine hydroxylase, the rate limiting enzyme in dopamine synthesis. No change in the expression of PPARγ on treatment with cocoa extract was observed when the cells were pre-treated with PPARγ antagonist GW9662. This data suggests that cocoa mediates mitochondrial biogenesis via a PPARγ/PGC1α dependent signalling pathway and also has the ability to improve dopaminergic functions by increasing tyrosine hydroxylase expression. Based on our data, we propose that a cocoa bean extract and products thereof could be used as potential nutritional supplements for neuroprotection in PD.

Spatial Variability of Aroma Profiles of Cocoa Trees Obtained through Computer Vision and Machine Learning Modelling: A Cover Photography and High Spatial Remote Sensing Application.

Cocoa is an important commodity crop, not only to produce chocolate, one of the most complex products from the sensory perspective, but one that commonly grows in developing countries close to the tropics. This paper presents novel techniques applied using cover photography and a novel computer application (VitiCanopy) to assess the canopy architecture of cocoa trees in a commercial plantation in Queensland, Australia. From the cocoa trees monitored, pod samples were collected, fermented, dried, and ground to obtain the aroma profile per tree using gas chromatography. The canopy architecture data were used as inputs in an artificial neural network (ANN) algorithm, with the aroma profile, considering six main aromas, as targets. The ANN model rendered high accuracy (correlation coefficient (R) = 0.82; mean squared error (MSE) = 0.09) with no overfitting. The model was then applied to an aerial image of the whole cocoa field studied to produce canopy vigor, and aroma profile maps up to the tree-by-tree scale. The tool developed could significantly aid the canopy management practices in cocoa trees, which have a direct effect on cocoa quality.

The Chemistry behind Chocolate Production.

Chocolate production is a complex process during which numerous chemical reactions occur. The most important processes, involving most of the reactions important for development of the proper chocolate flavor, are fermentation, drying and roasting of cocoa bean, and chocolate conching. During fermentation, formation of important precursors occurs, which are essential for further chemical reactions in the following processes of chocolate production. Roasting is one of the most important processes due to the occurrence of Maillard's reactions, during which aroma compounds are formed. In this paper, we have reviewed the most important chemical reactions that occur with proteins, carbohydrates, lipids, and polyphenols. Additionally, we present other components that may be naturally present or form during the production process, such as methylxanthines, aldehydes, esters, ketones, pyrazines, acids, and alcohols.

Characterization of the Proteome of Theobroma cacao Beans by Nano-UHPLC-ESI MS/MS.

Cocoa seed storage proteins play an important role in flavour development as aroma precursors are formed from their degradation during fermentation. Major proteins in the beans of Theobroma cacao are the storage proteins belonging to the vicilin and albumin classes. Although both these classes of proteins have been extensively characterized, there is still limited information on the expression and abundance of other proteins present in cocoa beans. This work is the first attempt to characterize the whole cocoa bean proteome by nano-UHPLC-ESI MS/MS analysis using tryptic digests of cocoa bean protein extracts. The results of this analysis show that >1000 proteins could be identified using a species-specific Theobroma cacao database. The majority of the identified proteins were involved with metabolism and energy. Additionally, a significant number of the identified proteins were linked to protein synthesis and processing. Several proteins were also involved with plant response to stress conditions and defence. Albumin and vicilin storage proteins showed the highest intensity values among all detected proteins, although only seven entries were identified as storage proteins. A comparison of MS/MS data searches carried out against larger non-specific databases confirmed that using a species-specific database can increase the number of identified proteins, and at the same time reduce the number of false positives. The results of this work will be useful in developing tools that can allow the comparison of the proteomic profile of cocoa beans from different genotypes and geographic origins. Data are available via ProteomeXchange with identifier PXD005586.

Dynamics and Biodiversity of Bacterial and Yeast Communities during Fermentation of Cocoa Beans.

Forastero hybrid cocoa bean fermentations have been carried out in a box (B) and in a heap (H), with or without the inoculation of Saccharomyces cerevisiae and Torulaspora delbrueckii as starter cultures. The bacteria, yeasts, and microbial metabolites (volatile and nonvolatile organic compounds) were monitored during fermentation to assess the connection between microbiota and the release of metabolites during this process. The presence of starter cultures was detected, by means of culture-dependent analysis, during the first 2 days of both fermentations. However, no statistical difference was observed in any of the physicochemical or microbiological analyses. Plate counts revealed the dominance of yeasts at the beginning of both fermentations, and these were followed by acetic acid bacteria (AAB) and lactic acid bacteria (LAB). Hanseniaspora opuntiae, S. cerevisiae, Pichia pijperi, Acetobacter pasteurianus, and Lactobacillus fermentum were the most abundant operational taxonomic units (OTUs) during both fermentation processes (B and H), although different relative abundances were observed. Only the diversity of the fungal species indicated a higher level of complexity in the B fermentations than in the H fermentations (P < 0.05), as well as a statistically significant difference between the initially inoculated starter cultures (P < 0.01). However, the microbial metabolite analysis indicated different distributions of the volatile and nonvolatile compounds between the two procedures, that is, B and H (P < 0.05), rather than between the inoculated and noninoculated fermentations. The box fermentations showed faster carbohydrate metabolism and greater production of organic acid compounds, which boosted the formation of alcohols and esters, than did the heap fermentations. Overall, the microbial dynamics and associations between the bacteria, yeasts, and metabolites were found to depend on the type of fermentation.IMPORTANCE In spite of the limited effectiveness of the considered inoculated starter strains, this study provides new information on the microbial development of box and heap cocoa fermentations, under inoculated and noninoculated conditions, as we coupled yeast/bacterial amplicon-based sequencing data with microbial metabolite detection. The information so far available suggests that microbial communities have played an important role in the evolution of aroma compounds. Understanding the pathways that microorganisms follow during the formation of aromas could be used to improve the fermentation processes and to enhance chocolate quality.

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.

Classification and compositional characterization of different varieties of cocoa beans by near infrared spectroscopy and multivariate statistical analyses.

Effective and fast methods are important for distinguishing cocoa varieties in the field and in the processing industry. This work proposes the application of NIR spectroscopy as a potential analytical method to classify different varieties and predict the chemical composition of cocoa. Chemical composition and colour features were determined by traditional methods and then related with the spectral information by partial least-squares regression. Several mathematical pre-processing methods including first and second derivatives, standard normal variate and multiplicative scatter correction were applied to study the influence of spectral variations. The results of chemical composition analysis and colourimetric measurements show significant differences between varieties. NIR spectra of samples exhibited characteristic profiles for each variety and principal component analysis showed different varieties in according to spectral features.

Clovamide and Its Derivatives-Bioactive Components of Theobroma cacao and Other Plants in the Context of Human Health.

Clovamide (N-caffeoyl-L-3,4-dihydroxyphenylalanine, N-caffeoyldopamine, N-caffeoyl-L-DOPA) is a derivative of caffeic acid, belonging to phenolamides (hydroxycinnamic acid amides). Despite a growing interest in the biological activity of natural polyphenolic substances, studies on the properties of clovamide and related compounds, their significance as bioactive components of the diet, as well as their effects on human health are a relatively new research trend. On the other hand, in vitro and in vivo evidence indicates the considerable potential of these substances in the context of maintaining human health or using them as pharmacophores. The name "clovamide" directly derives from red clover (Trifolium pratense L.), being the first identified source of this compound. In the human diet, clovamides are mainly present in chocolate and other cocoa-containing products. Furthermore, their occurrence in some medicinal plants has also been confirmed. The literature reports deal with the antioxidant, anti-inflammatory, neuroprotective, antiplatelet/antithrombotic and anticancer properties of clovamide-type compounds. This narrative review summarizes the available data on the biological activity of clovamides and their potential health-supporting properties, including prospects for the use of these compounds for therapeutic purposes.

Effect of Pod Storage and Drying Temperature on Fermentation Dynamics and Final Bean Quality of Cacao Nacional in Ecuador.

The impact of pod storage (PS) and two drying temperatures of fermented cocoa beans was investigated in Ecuador. Therefore, four variations were simultaneously carried out three times at two locations, independently: 0, 3, and 5 days of PS, dried at 60 °C and 0 days of PS, dried at 80 °C. Pod weight during storage, pulp content, pH, temperature, microbial counts, total free amino acids, protein profiles, sugars, organic acids, cut-test, fermentation index, and sensory profiles were analyzed. Minor differences in fermentation dynamics and bean quality were found between variations with and without PS. A rather accelerated fermentation with pod-stored beans was observed (e.g., faster color change, slightly lower pH in cotyledon after 48 h), along with a significantly higher maximal temperature during 24-42 h (43.1 ± 3.2 °C compared to 39.2 ± 2.0 °C without PS). More well-fermented beans were reached with PS (52.3 ± 22.6%) than without (62.7 ± 9.2%). Differences during fermentation were observed between the locations (e.g., pH, acids, sugars), but sensory evaluation indicated that the impact of location was mitigated with PS. Drying at 80 °C showed no adverse effects, as evidenced by the results of the cut-test and fermentation index. However, sensory evaluations revealed significant differences between 80 °C and 60 °C, with the former exhibiting more bitter and astringent cocoa liquor.

Simultaneous discovery of compounds dominated by either molding kinetics or geographical region of origin for moisture damaged cacao beans using orthogonally applied tile-based fisher ratio analysis of GC×GC-TOFMS data.

Herein, two "orthogonal" characteristics of moisture damaged cacao beans (temporally dependent molding kinetics versus the time-independent geographical region of origin) are simultaneously analyzed in a comprehensive two-dimensional (2D) gas chromatography time-of-flight mass spectrometry (GC×GC-TOFMS) dataset using tile-based Fisher ratio (F-ratio) analysis. Cacao beans from six geographical regions were analyzed once a day for six days following the initiation of moisture damage to trigger the molding process. Thus, there are two "extremes" to the experimental sample class design: six time points for the molding kinetics versus the six geographical regions of origin, resulting in a 6 × 6 element signal array referred to as a composite chemical fingerprint (CCF) for each analyte. Usually, this study would involve initial generation of two separate hit lists using F-ratio analysis, one hit list from inputting the data with the six time point classes, then another hit list from inputting the dataset from the perspective of geographic region of origin. However, analysis of two separate hit lists with the intent to distill them down to one hit list is extremely time-consuming and fraught with shortcomings due to the challenges associated with attempting to match analytes across two hit lists. To address this challenge, tile-based F-ratio analysis is "orthogonally applied" to each analyte CCF to simultaneously determine two F-ratios at the chromatographic 2D location (F-ratiokinetic and F-ratioregion) for each hit, by ranking a single hit list using the higher of the two F-ratios resulting in the discovery of 591 analytes. Further, using a pseudo-null distribution approach, at the 99.9% threshold over 400 analytes were deemed suitable for PCA classification. Using a more stringent 99.999% threshold, over 100 analytes were explored more deeply using PARAFAC to provide a purified mass spectrum.

Influences of Depulping, Pod Storage and Fermentation Time on Fermentation Dynamics and Quality of Ghanaian Cocoa.

This study investigated the impact of the depulping of cocoa beans after pod opening, as well as the influences of pod storage (PS) and fermentation time on the fermentation dynamics and the overall quality of beans and liquors made thereof. Twelve variations were conducted in three experimental runs (with/without depulping; 1-/3-day PS; and fermentation times of 3, 4, 5, 6 or 7 days). Fermentation dynamics (e.g., temperature and pH) and the quality of dried beans (e.g., cut-test and fermentation index) and liquors (sensory assessment, quantification of cocoa key-odorants and tastants) were investigated. It was demonstrated that 17-20% of cocoa pulp, relative to the total bean-pulp-mass weight, could be mechanically removed without negatively affecting the bean quality. No significant differences were found in the percentages of well-fermented beans after 5-6 days fermentation with 1-day PS, resulting in 49 ± 9% with, and 48 ± 12% without depulping. There were no significant differences in key tastants present in the liquors; however, significantly less volatile acids and esters were found when liquors were produced from 5-6 day-fermented depulped beans, with 1-day PS, without negatively affecting the sensory profiles. This strategy allows producers to maximize the cacao fruit's value by integrating part of the pulp into the cocoa value chain.