Evaluation of Cocoa Bean Shell Antimicrobial Activity: A Tentative Assay Using a Metabolomic Approach for Active Compound Identification.

Cocoa bean shell is one of the main by-products of chocolate manufacturing and possesses several compounds with biofunctionalities. It can function as an antibacterial agent, and its action is mostly reported against Streptococcus mutans. However, only a few studies have investigated the cocoa bean shell compounds responsible for this activity. This study aimed to evaluate several extracts of cocoa bean shells from different geographical origins and cocoa varieties and estimate their antimicrobial properties against different fungal and bacterial strains by determining their minimal inhibitory concentration. The results demonstrated antimicrobial activity of cocoa bean shell against one of the tested strains, S. mutans. Cocoa bean shell extracts were further analysed via LC-HRMS for untargeted metabolomic analysis. LC-HRMS data were analysed (preprocessing and statistical analyses) using the Workflow4Metabolomics platform. The latter enabled us to identify possible compounds responsible for the detected antimicrobial activity by comparing the more and less active extracts. Active extracts were not the most abundant in polyphenols but contained higher concentrations of two metabolites. After tentative annotation of these metabolites, one of them was identified and confirmed to be 7-methylxanthine. When tested alone, 7-methylxanthine did not display antibacterial activity. However, a possible cocktail effect due to the synergistic activity of this molecule along with other compounds in the cocoa bean shell extracts cannot be neglected. In conclusion, cocoa bean shell could be a functional ingredient with benefits for human health as it exhibited antibacterial activity against S. mutans. However, the antimicrobial mechanisms still need to be confirmed.

Characterization and Classification of Cocoa Bean Shells from Different Regions of Venezuela Using HPLC-PDA-MS/MS and Spectrophotometric Techniques Coupled to Chemometric Analysis.

The cocoa bean shell (CBS) is one of the main cocoa byproducts with a prospective to be used as a functional food ingredient due to its nutritional and sensory properties. This study aims to define the chemical fingerprint of CBSs obtained from cocoa beans of diverse cultivars and collected in different geographical areas of Venezuela assessed using high-performance liquid chromatography coupled to photodiodes array and mass spectrometry (HPLC-PDA-MS/MS) and spectrophotometric assays combined with multivariate analysis for classification purposes. The study provides a comprehensive fingerprint and quantitative data for 39 compounds, including methylxanthines and several polyphenols, such as flavan-3-ols, procyanidins, and N-phenylpropenoyl amino acids. Several key cocoa markers, such as theobromine, epicatechin, quercetin-3-O-glucoside, procyanidin_A pentoside_3, and N-coumaroyl-l-aspartate_2, were found suitable for the classification of CBS according to their cultivar and origin. Despite the screening methods required a previous purification of the sample, both methodologies appear to be suitable for the classification of CBS with a high correlation between datasets. Finally, preliminary findings on the identification of potential contributors for the radical scavenging activity of CBS were also accomplished to support the valorization of this byproduct as a bioactive ingredient in the production of functional foods.

Chemometric Classification of Cocoa Bean Shells Based on Their Polyphenolic Profile Determined by RP-HPLC-PDA Analysis and Spectrophotometric Assays.

The cocoa bean shell (CBS), a byproduct from the cocoa industry, was recently proposed as a functional and low-cost ingredient, mainly because of its content in polyphenols. However, vegetal food products could significantly differ in their chemical composition depending on different factors such as their geographical provenience. This work is aimed to determine the polyphenolic and methylxanthine profile of different CBS samples and utilize it for achieving their differentiation according to their geographical origin and variety. RP-HPLC-PDA was used to determine the CBS polyphenolic profile. Spectrophotometric assays were used to obtain the total phenolic, flavonoid, and tannin contents, as well as to evaluate their radical scavenging activity. The results obtained from both methods were then compared and used for the CBS differentiation according to their origin and varieties through chemometric analysis. RP-HPLC-PDA allowed to determine 25 polyphenolic compounds, as well as the methylxanthines theobromine and caffeine. Polyphenolic profile results highlighted significant differences among the analyzed samples, allowing for their differentiation based on their geographical provenience. Similar results were achieved with the results of the spectrophotometric assays, considered as screening methods. Differentiation based on CBS variety was instead obtained based on the HPLC-determined methylxanthine profile.

Cocoa Bean Shell-A By-Product with Nutritional Properties and Biofunctional Potential.

Cocoa bean shells (CBS) are one of the main by-products from the transformation of cocoa beans, representing 10%‒17% of the total cocoa bean weight. Hence, their disposal could lead to environmental and economic issues. As CBS could be a source of nutrients and interesting compounds, such as fiber (around 50% w/w), cocoa volatile compounds, proteins, minerals, vitamins, and a large spectrum of polyphenols, CBS may be a valuable ingredient/additive for innovative and functional foods. In fact, the valorization of food by-products within the frame of a circular economy is becoming crucial due to economic and environmental reasons. The aim of this review is to look over the chemical and nutritional composition of CBS and to revise the several uses that have been proposed in order to valorize this by-product for food, livestock feed, or industrial usages, but also for different medical applications. A special focus will be directed to studies that have reported the biofunctional potential of CBS for human health, such as antibacterial, antiviral, anticarcinogenic, antidiabetic, or neuroprotective activities, benefits for the cardiovascular system, or an anti-inflammatory capacity.

Physical Properties and Consumer Evaluation of Cocoa Bean Shell-Functionalized Biscuits Adapted for Diabetic Consumers by the Replacement of Sucrose with Tagatose.

The cocoa bean shell (CBS), a by-product of the cocoa industry, has been reported to be rich in fiber and polyphenols, which could contribute to reducing the metabolism of sugars and glucose adsorption. The production of CBS-based biscuits in which sucrose is replaced with tagatose (a low-glycemic sugar with prebiotic properties), benefiting diabetic consumers, is proposed. Six prototype biscuits were produced using sucrose, tagatose, and CBS powder at 0%, 10%, and 20% as a wheat flour replacement. Biscuits were studied in terms of fiber content, and those with 10% and 20% CBS showed to contain 5.66% and 8.70-8.71% of total dietary fiber, respectively. Moreover, the physicochemical and structural properties of the biscuits were studied to evaluate their differences due to the use of sucrose and tagatose combined with CBS. Significant effects mainly caused by the reducing nature and lower solubility of tagatose with respect to sugar, and the water retention capacities of CBS were observed. Finally, the biscuits were evaluated by performing a consumer acceptance evaluation, and their perceptible sensorial differences were studied by performing a Napping® sensory characterization. CBS-based biscuits represent an interesting possibility for cocoa by-product revalorization, although an optimized recipe is recommended, especially when employing tagatose.

Polyphenolic and Methylxanthine Bioaccessibility of Cocoa Bean Shell Functional Biscuits: Metabolomics Approach and Intestinal Permeability through Caco-2 Cell Models.

Cocoa bean shell (CBS), a by-product with considerable concentrations of bioactive compounds and proven biofunctional potential, has been demonstrated to be a suitable ingredient for high-fiber functional biscuits adapted to diabetic consumers. In this work, the in vitro bioaccessibility and intestinal absorption of polyphenols and methylxanthines contained in these biscuits were evaluated, and the effect of the food matrix was studied. Biscuits containing CBS and the CBS alone underwent in vitro digestion followed by an intestinal permeability study. The results confirmed that compounds were less bioavailable in the presence of a food matrix, although the digestion contributed to their release from this matrix, increasing the concentrations available at the intestinal level and making them capable of promoting antioxidant and antidiabetic activities. After digestion, CBS biscuits were shown to possess α-glucosidase inhibition capacity comparable to that of acarbose. Moreover, the presence of the food matrix improved the stability of polyphenols throughout the digestion process. Intestinal absorption of flavan-3-ols seemed to be limited to a maximum threshold and was therefore independent of the sample, while procyanidin was not absorbed. Methylxanthine absorption was high and was boosted by the presence of the food matrix. The results confirmed the biofunctional potential of CBS-based biscuits.

In Vitro Bioaccessibility and Functional Properties of Phenolic Compounds from Enriched Beverages Based on Cocoa Bean Shell.

The cocoa bean shell (CBS), a cocoa by-product, contains a significant number of bioactive compounds with functional properties, such as polyphenols and methylxanthines, and is used as an ingredient in beverages and foods. In this work, the bioaccessibility of polyphenols and methylxanthines after In Vitro digestion was evaluated in new flavoured beverages for at-home consumption (capsules and tea bags). In addition, the polyphenolic composition, functional properties (antiradical and α-glucosidase inhibition capacities) and consumer acceptability of these beverages were evaluated. In both capsule and tea bag beverages, the bioaccessibility of methylxanthines was 100% while that of total polyphenols exceeded 50%. The main polyphenols determined using reverse-phase liquid chromatography were type B procyanidins and epicatechin. The antiradical activity in capsule and tea bag beverages was 1.75 and 1.88 mM of Trolox equivalents, respectively, of which 59.50% and 57.09% were recovered after simulated digestion. The percentage of α-glucosidase inhibition before In Vitro digestion (51.64% and 53.82% for capsules and tea bags, respectively) was comparable to that of acarbose at 0.5 mM. All the beverages obtained a high consumer acceptability. Therefore, these results highlight that CBSs can be used as a valid source of bioactive compounds in the preparation of beverages with homemade techniques.

Assessment of volatile fingerprint by HS-SPME/GC-qMS and E-nose for the classification of cocoa bean shells using chemometrics.

The cocoa bean shell (CBS) is a main by-product of cocoa processing, with great potential to be used as an ingredient for functional foods because of its nutritional and flavour properties. This study aimed to characterise and classify CBSs obtained from cocoa beans of diverse cultivars and collected in different geographical origins through their volatile profile assessed using headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME/GC-qMS) and E-nose combined with principal component analysis (PCA). The study provides, for the first time in a representative set of samples, a comprehensive fingerprint and semi-quantitative data for >100 volatile organic compounds (VOCs), such as aldehydes, ketones, pyrazines, alcohols, and acids. Through PCA, a clear separation of the Criollo cultivar from the other cultivars was achieved with both GC-qMS and E-nose analytical techniques because of the high content of key-aroma VOCs. Several biomarkers identified by GC-qMS, such as 2-hepanol, 2-methylpropanoic acid, and 2,3,5-trimethylpyrazine, recognized as key-aroma compounds for cocoa beans, were found suitable for the classification of CBSs according to their quality and origin. GC-qMS and E-nose appeared to be suitable analytical approaches to classify CBSs, with a high correlation between both analytical techniques. The volatile fingerprint and classification of CBSs could allow for the selection of samples with a specific flavour profile according to the food application and, therefore, constitute an interesting approach to valorise this by-product as a food ingredient.

Analytical dataset on volatile compounds of cocoa bean shells from different cultivars and geographical origins.

This data article describes the analysis of volatile organic compounds (VOCs) in 44 samples of cocoa bean shells (CBS) obtained from cocoa beans of diverse cultivars and collected in different geographical origins. The volatile compounds were extracted by headspace solid-phase microextraction (HS-SPME) method and then analyzed by gas chromatography coupled to a quadrupole mass spectrometry GC-qMS. The retention times, identification and semi-quantification of 101 VOCs are reported. Data collected on the volatile profile of CBS samples using E-nose analysis are also available. Additional data related to physicochemical characteristics and color analysis for CBS samples are reported. Further interpretation and discussion on these datasets can be found in the related article entitled "Assessment of volatile fingerprint by HS-SPME/GC-qMS and E-nose for the classification of cocoa bean shells using chemometrics" (Barbosa-Pereira et al., 2019).

Effects of Particle Size and Extraction Methods on Cocoa Bean Shell Functional Beverage.

One of the main by-products in cocoa industry is the cocoa bean shell (CBS), which represents approximately 12-20% of the bean. This product has been suggested as a food ingredient because of its aroma and high dietary fiber and polyphenol contents. The purpose of this work was to evaluate the effects of the CBS particle size and extraction methods on the chemical composition and consumer acceptance of a functional beverage, in order to find the best combination of technological parameters and health benefits. Five particle sizes of CBS powder and six home techniques were used for beverage preparation. The influence of these factors on the physico-chemical characteristics, methylxanthine and polyphenolic contents, antioxidant and antidiabetic properties, and consumer acceptance was evaluated. Total phenolic content values up to 1803.83 mg GAE/L were obtained for the beverages. Phenolic compounds and methylxanthines were identified and quantified by HPLC-PDA. These compounds may be related to the high antioxidant capacity (up to 7.29 mmol TE/L) and antidiabetic properties (up to 52.0% of α-glucosidase inhibition) observed. Furthermore, the consumer acceptance results indicated that CBS may represent an interesting ingredient for new functional beverages with potential health benefits, reducing the environmental and economic impact of by-product disposal.