Bio-Composite Filaments Based on Poly(Lactic Acid) and Cocoa Bean Shell Waste for Fused Filament Fabrication (FFF): Production, Characterization and 3D Printing.

In this study, novel biocomposite filaments incorporating cocoa bean shell waste (CBSW) and poly(lactic acid) (PLA) were formulated for application in Fused Filament Fabrication (FFF) technology. CBSW, obtained from discarded chocolate processing remnants, was blended with PLA at concentrations of 5 and 10 wt.% to address the challenge of waste material disposal while offering eco-friendly composite biofilaments for FFF, thereby promoting resource conservation and supporting circular economy initiatives. A comprehensive analysis encompassing structural, morphological, thermal, and mechanical assessments of both raw materials and resultant products (filaments and 3D printed bars) was conducted. The findings reveal the presence of filler aggregates only in high concentrations of CBSW. However, no significant morphological or thermal changes were observed at either CBSW concentration (5 wt.% and 10 wt.%) and satisfactory printability was achieved. In addition, tensile tests on the 3D printed objects showed improved stiffness and load resistance in these samples at the highest CBSW concentrations. In addition, to demonstrate their practical application, several 3D prototypes (chocolate-shaped objects) were printed for presentation in the company's shop window as a chocolate alternative; while retaining the sensory properties of the original cocoa, the mechanical properties were improved compared to the base raw material. Future research will focus on evaluating indicators relevant to the preservation of the biocomposite's sensory properties and longevity.

Nanocellulose from Cocoa Shell in Pickering Emulsions of Cocoa Butter in Water: Effect of Isolation and Concentration on Its Stability and Rheological Properties.

There is a growing interest in developing new strategies to completely or partially replace cocoa butter in food and cosmetic products due to its cost and health effects. One of these alternatives is to develop stable emulsions of cocoa butter in water. However, incorporating cocoa butter is challenging as it solidifies and forms crystals, destabilizing the emulsion through arrested coalescence. Prevention against this destabilization mechanism is significantly lower than against coalescence. In this research, the rheological properties of nanocellulose from cocoa shell, a by-product of the chocolate industry, were controlled through isolation treatments to produce nanocellulose with a higher degree of polymerization (DP) and a stronger three-dimensional network. This nanocellulose was used at concentrations of 0.7 and 1.0 wt %, to develop cocoa butter in-water Pickering emulsion using a high shear mixing technique. The emulsions remained stable for more than 15 days. Nanocellulose was characterized using attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), hot water and organic extractives, atomic force microscopy (AFM), degree of polymerization (DP), and rheological analysis. Subsequently, the emulsions were characterized on days 1 and 15 after their preparation through photographs to assess their physical stability. Fluorescent and electronic microscopy, as well as rheological analysis, were used to understand the physical properties of emulsions.

Cocoa Shell Extract Reduces Blood Pressure in Aged Hypertensive Rats via the Cardiovascular Upregulation of Endothelial Nitric Oxide Synthase and Nuclear Factor (Erythroid-Derived 2)-like 2 Protein Expression.

Cocoa shell is a by-product of cocoa manufacturing. We obtained an aqueous extract (CSE) rich in polyphenols and methylxanthines with antioxidant and vasodilatory properties. We aimed to evaluate the effects of CSE supplementation in aged hypertensive rats on blood pressure and the mechanism implicated. Eighteen-month-old male and female rats exposed to undernutrition during the fetal period who developed hypertension, with a milder form in females, were used (MUN rats). Systolic blood pressure (SBP; tail-cuff plethysmography) and a blood sample were obtained before (basal) and after CSE supplementation (250 mg/kg; 2 weeks, 5 days/week). Plasma SOD, catalase activity, GSH, carbonyls, and lipid peroxidation were assessed (spectrophotometry). In hearts and aortas from supplemented and non-supplemented age-matched rats, we evaluated the protein expression of SOD-2, catalase, HO-1, UCP-2, total and phosphorylated Nrf2 and e-NOS (Western blot), and aorta media thickness (confocal microscopy). MUN males had higher SBP compared with females, which was reduced via CSE supplementation with a significant difference for group, sex, and interaction effect. After supplementation with plasma, GSH, but not catalase or SOD, was elevated in males and females. Compared with non-supplemented rats, CSE-supplemented males and females exhibited increased aorta e-NOS and Nrf2 protein expression and cardiac phosphorylated-Nrf2, without changes in SOD-2, catalase, HO-1, or UCP-2 in cardiovascular tissues or aorta remodeling. In conclusion, CSE supplementation induces antihypertensive actions related to the upregulation of e-NOS and Nrf2 expression and GSH elevation and a possible direct antioxidant effect of CSE bioactive components. Two weeks of supplementation may be insufficient to increase antioxidant enzyme expression.

Cocoa Shell Infusion: A Promising Application for Added-Value Beverages Based on Cocoa's Production Coproducts.

The cocoa shell (CS) is being incorporated into different food products due to its recognized content of bioactive compounds. In the case of cocoa shell infusions (CSI), the bioactive compounds that manage to be transferred to the infusion have yet to be clearly known, i.e., what is really available to the consumer. In this study, CS was obtained from toasted Colombian Criollo cocoa beans. Three particle sizes (A: >710 µm; B: >425 and <710 µm; C: <425 µm) were evaluated in the CSI, which was traditionally prepared by adding CS to hot water (1%). The decrease in particle size increased the antioxidant capacity (DPPH and ABTS) and the total phenolic compounds. A significant effect (p < 0.05) both of the particle size and of the temperature of tasting was found on some sensory attributes: greater bitterness, acidity, and astringency were due to the greater presence of epicatechin, melanoidins, and proanthocyanidins in the smaller particle sizes. The analysis of the volatile organic compounds showed that the CSI aroma was characterized by the presence of nonanal, 2-nonanone, tetramethylpyrazine, α-limonene, and linalool, which present few variations among the particle sizes. Moreover, analysis of biogenic amines, ochratoxin A, and microbial load showed that CSI is not a risk to public health. Reducing particle size becomes an important step to valorize the functional properties of CS and increase the quality of CSI.

Effect of Supplementation with Coffee and Cocoa By-Products to Ameliorate Metabolic Syndrome Alterations Induced by High-Fat Diet in Female Mice.

Coffee and cocoa manufacturing produces large amounts of waste. Generated by-products contain bioactive compounds with antioxidant and anti-inflammatory properties, suitable for treating metabolic syndrome (MetS). We aimed to compare the efficacy of aqueous extracts and flours from coffee pulp (CfPulp-E, CfPulp-F) and cocoa shell (CcShell-E, CcShell-F) to ameliorate MetS alterations induced by a high-fat diet (HFD). Bioactive component content was assessed by HPLC/MS. C57BL/6 female mice were fed for 6 weeks with HFD followed by 6 weeks with HFD plus supplementation with one of the ingredients (500 mg/kg/day, 5 days/week), and compared to non-supplemented HFD and Control group fed with regular chow. Body weight, adipocyte size and browning (Mitotracker, confocal microscopy), plasma glycemia (basal, glucose tolerance test-area under the curve, GTT-AUC), lipid profile, and leptin were compared between groups. Cocoa shell ingredients had mainly caffeine, theobromine, protocatechuic acid, and flavan-3-ols. Coffee pulp showed a high content in caffeine, protocatechuic, and chlorogenic acids. Compared to Control mice, HFD group showed alterations in all parameters. Compared to HFD, CcShell-F significantly reduced adipocyte size, increased browning and high-density lipoprotein cholesterol (HDL), and normalized basal glycemia, while CcShell-E only increased HDL. Both coffee pulp ingredients normalized adipocyte size, basal glycemia, and GTT-AUC. Additionally, CfPulp-E improved hyperleptinemia, reduced triglycerides, and slowed weight gain, and CfPulp-F increased HDL. In conclusion, coffee pulp ingredients showed a better efficacy against MetS, likely due to the synergic effect of caffeine, protocatechuic, and chlorogenic acids. Since coffee pulp is already approved as a food ingredient, this by-product could be used in humans to treat obesity-related MetS alterations.

Radical Scavenging and Cellular Antioxidant Activity of the Cocoa Shell Phenolic Compounds after Simulated Digestion.

The cocoa industry generates a considerable quantity of cocoa shell, a by-product with high levels of methylxanthines and phenolic compounds. Nevertheless, the digestion process can extensively modify these compounds' bioaccessibility, bioavailability, and bioactivity as a consequence of their transformation. Hence, this work's objective was to assess the influence of simulated gastrointestinal digestion on the concentration of phenolic compounds found in the cocoa shell flour (CSF) and the cocoa shell extract (CSE), as well as to investigate their radical scavenging capacity and antioxidant activity in both intestinal epithelial (IEC-6) and hepatic (HepG2) cells. The CSF and the CSE exhibited a high amount of methylxanthines (theobromine and caffeine) and phenolic compounds, mainly gallic acid and (+)-catechin, which persisted through the course of the simulated digestion. Gastrointestinal digestion increased the antioxidant capacity of the CSF and the CSE, which also displayed free radical scavenging capacity during the simulated digestion. Neither the CSF nor the CSE exhibited cytotoxicity in intestinal epithelial (IEC-6) or hepatic (HepG2) cells. Moreover, they effectively counteracted oxidative stress triggered by tert-butyl hydroperoxide (t-BHP) while preventing the decline of glutathione, thiol groups, superoxide dismutase, and catalase activities in both cell lines. Our study suggests that the cocoa shell may serve as a functional food ingredient for promoting health, owing to its rich concentration of antioxidant compounds that could support combating the cellular oxidative stress associated with chronic disease development.

Changes in the cocoa shell dietary fiber and phenolic compounds after extrusion determine its functional and physiological properties.

The influence of different extrusion conditions on the cocoa shell (CS) dietary fiber, phenolic compounds, and antioxidant and functional properties was evaluated. Extrusion produced losses in the CS dietary fiber (3-26%), especially in the insoluble fraction, being more accentuated at higher temperatures (160 °C) and lower moisture feed (15-20%). The soluble fiber fraction significantly increased at 135 °C because of the solubilization of galactose- and glucose-containing insoluble polysaccharides. The extruded CS treated at 160 °C-25% of feed moisture showed the highest increase of total (27%) and free (58%) phenolic compounds, accompanied by an increase of indirect (10%) and direct (77%) antioxidant capacity. However, more promising results relative to the phenolic compounds' bioaccessibility after in vitro simulated digestion were observed for 135°C-15% of feed moisture extrusion conditions. The CS' physicochemical and techno-functional properties were affected by extrusion, producing extrudates with higher bulk density, a diminished capacity to hold oil (22-28%) and water (18-65%), and improved swelling properties (14-35%). The extruded CS exhibited increased glucose adsorption capacity (up to 2.1-fold, at 135 °C-15% of feed moisture) and α-amylase in vitro inhibitory capacity (29-54%), accompanied by an increase in their glucose diffusion delaying ability (73-91%) and their starch digestion retardation capacity (up to 2.8-fold, at 135 °C-15% of feed moisture). Moreover, the extruded CS preserved its cholesterol and bile salts binding capacity and pancreatic lipase inhibitory properties. These findings generated knowledge of the CS valorization through extrusion to produce foods rich in dietary fiber with improved health-promoting properties due to the extrusion-triggered fiber solubilization.

Handheld and benchtop vis/NIR spectrometer combined with PLS regression for fast prediction of cocoa shell in cocoa powder.

The fermented and dried cocoa beans are peeled, either before or after the roasting process, as peeled nibs are used for chocolate production, and shell content in cocoa powders may result from economically motivated adulteration (EMA), cross-contamination or misfits in equipment in the peeling process. The performance of this process is carefully evaluated, as values above 5% (w/w) of cocoa shell can directly affect the sensory quality of cocoa products. In this study chemometric methods were applied to near-infrared (NIR) spectra from a handheld (900-1700 nm) and a benchtop (400-1700 nm) spectrometers to predict cocoa shell content in cocoa powders. A total of 132 binary mixtures of cocoa powders with cocoa shell were prepared at several proportions (0 to 10% w/w). Partial least squares regression (PLSR) was used to develop the calibration models and different spectral preprocessing were investigated to improve the predictive performance of the models. The ensemble Monte Carlo variable selection (EMCVS) method was used to select the most informative spectral variables. Based on the results obtained with both benchtop (R2P = 0.939, RMSEP = 0.687% and RPDP = 4.14) and handheld (R2P = 0.876, RMSEP = 1.04% and RPDP = 2.82) spectrometers, NIR spectroscopy combined with the EMCVS method proved to be a highly accurate and reliable tool for predicting cocoa shell in cocoa powder. Even with a lower predictive performance than the benchtop spectrometer, the handheld spectrometer has potential to specify whether the amount of cocoa shell present in cocoa powders is in accordance with the Codex Alimentarius specifications.

Ultrasound-Assisted Extraction of Bioactive Compounds from Cocoa Shell and Their Encapsulation in Gum Arabic and Maltodextrin: A Technology to Produce Functional Food Ingredients.

In this study, the extraction of cocoa shell powder (CSP) was optimized, and the optimized extracts were spray-dried for encapsulation purposes. Temperature (45−65 °C), extraction time (30−60 min), and ethanol concentration (60−100%) were the extraction parameters. The response surface methodology analysis revealed that the model was significant (p ≤ 0.05) in interactions between all variables (total phenolic compound, total flavonoid content, and antioxidant activity as measured by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP assays), with a lack of fit test for the model being insignificant (p > 0.05). Temperature (55 °C), time (45 min), and ethanol concentration (60%) were found to be the optimal extraction conditions. For spray-drying encapsulation, some quality metrics (e.g., water solubility, water activity) were insignificant (p > 0.05). The microcapsules were found to be spherical in shape using a scanning electron microscope. Thermogravimetric and differential thermogravimetric measurements of the microcapsules revealed nearly identical results. The gum arabic + maltodextrin microcapsule (GMM) showed potential antibacterial (zone of inhibition: 11.50 mm; lower minimum inhibitory concentration: 1.50 mg/mL) and antioxidant (DPPH: 1063 mM trolox/100g dry wt.) activities (p ≤ 0.05). In conclusion, the microcapsules in this study, particularly GMM, are promising antioxidant and antibacterial agents to be fortified as functional food ingredients for the production of nutraceutical foods with health-promoting properties.

Gastrointestinal fate of phenolic compounds and amino derivatives from the cocoa shell: An in vitro and in silico approach.

The objective of this study was to assess how in vitro gastrointestinal digestion influenced the bioaccessibility and potential bioavailability of phenolic compounds and methylxanthines in thecocoa shell (CS) in the form of flour (CSF) and aqueous extract (CSE). To comprehend how these phytochemicals behaved during gastrointestinal digestion, we also modeled in silico the colonic microbial biotransformation of the phenolic compounds in the CS. Different groups of phenolic compounds (mainly gallic andprotocatechuic acids, and catechin) and methylxanthines (theobromine and caffeine)could be found in the CS. Methylxanthines and phenolic compounds were released differently during gastrointestinal digestion. Whereas digestion triggered the release of hydroxybenzoic acids (67-73%) and flavan-3-ols (73-88%) during the intestinal phase, it also caused the degradation of flavonols and flavones. Besides, the release of phytochemicals was significantly influenced by the CS matrix type. Phenolic compounds were protected by the CSF matrix. Phenolic acids from CSF were more bioaccessible in the intestinal (1.2-fold, p < 0.05) and colonic (1.3-fold, p < 0.05) phases than those from the CSE. Methylxanthines were also more bioaccessible in the intestinal (1.8-fold, p < 0.01) and colonic phases (1.3-fold, p < 0.001) and bioavailable (1.8-fold, p < 0.001) in the CSF. Colonic metabolism demonstrated that the gut microbiota could biotransform non-absorbed phenolic compounds into other lower molecular weight and more bioavailable metabolites. These findings support the CS's potential as a source of bioaccessible, bioavailable, and active phytochemicals.