Cocoa extract induces browning of white adipocytes and improves glucose intolerance in mice fed a high-fat diet.

Cocoa extract (CE) offers several health benefits, such as anti-obesity and improved glucose intolerance. However, the mechanisms remain unclear. Adipose tissue includes white adipose tissue (WAT) and brown adipose tissue. Brown adipose tissue leads to body fat reduction by metabolizing lipids to heat via uncoupling protein 1 (UCP1). The conversion of white adipocytes into brown-like adipocytes (beige adipocytes) is called browning, and it contributes to the anti-obesity effect and improved glucose tolerance. This study aimed to evaluate the effect of CE on glucose tolerance in terms of browning. We found that dietary supplementation with CE improved glucose intolerance in mice fed a high-fat diet, and it increased the expression levels of Ucp1 and browning-associated gene in inguinal WAT. Furthermore, in primary adipocytes of mice, CE induced Ucp1 expression through ß3-adrenergic receptor stimulation. These results suggest that dietary CE improves glucose intolerance by inducing browning in WAT.

Influence of cocoa products on rheological, fatty acid profile and quality characteristics of biscuits.

Cocoa products namely cocoa powder (CP), cocoa butter (CB) and cocoa mass (CM) were selected for their utilization in soft dough biscuits. CP was blended with the refined wheat flour (WF-0, 5, 10 and 15% levels) and rheological and quality characteristics of biscuits were studied. The spread ratio decreased (10.1-8.8), density (0.49-0.52 g/cm3) and breaking strength values (1127-1369 g force) increased gradually with increase in CP. Combination of GMS and SSL at 0.25% each improved the quality of biscuits at 10% incorporation of CP. Further the biscuit fat (BF) was replaced with CB (0, 25, 50 and 75%). Later the biscuits with CM were prepared by replacing the flour (15%) and BF (0, 25, 50 and 75%). Acceptability of the CM based biscuits was better when compared to CB based biscuits. The total polyphenol content in control biscuits was 55.55 mg/100 g and was in the range between 81.98 and 102.05 mg/100 g for cocoa based biscuits. The protein content in cocoa based biscuits was marginally higher than the control biscuit. Though there was a wide variation in the fat content and different fatty acids in raw materials, interestingly, the values varied narrowly in biscuits.

Investigating Key Volatile Compound Diffusion in Cocoa Beans during Yeast Fermentation-like Incubation.

An experimental setup was devised to investigate the permeability of cocoa bean seed coat and pulp to key volatile compounds during fermentation. Four labeled compounds (ethyl acetate-d3, ethyl octanoate-d15, 2-phenylethanol-d5, linalool-d5) and 2 unlabeled (beta-damascenone, delta-decalactone) were chosen for the investigation. The beans (cotyledons), depulped beans, or pulped beans were immersed separately in a concentrated solution of these volatile compounds at 36 or 46 °C for durations ranging from 3 to 120 h. The imbibed beans were dissected, and the cotyledons were analyzed by SPME-GC/MS. The diffusion of volatile compounds from the external solution to the seed was categorized into three groups: (1) not diffusible (ethyl octanoate-d15); (2) semidiffusible (ethyl acetate); and (3) totally diffusible (2-phenylethanol-d5, linalool-d5, beta-damascenone, delta-decalactone). The impact of the yeast on volatile compound diffusion was also investigated by immerging the pulped beans into the same concentrated solution with a yeast starter. Results highlighted the positive role of yeast in the diffusion of volatile compounds. The starter positively contributed to volatile compound diffusion after a transition phase occurring at approximately 48 h of fermentation, enriching the cocoa beans with key aromatic volatile compounds.

Salmonella prevalence in raw cocoa beans and a microbiological risk assessment to evaluate the impact of cocoa liquor processing on the reduction of Salmonella.

Salmonella in raw cocoa beans (n= 870) from main sourcing areas over nine months was analyzed. It was detected in 71 (ca. 8.2%) samples, with a contamination level of 0.3-46 MPN/g except for one sample (4.1×104 CFU/g). Using prevalence and concentration data as input, the impact of thermal treatment in cocoa processing on the risk estimate of acquiring salmonellosis by a random Belgian chocolate consumer was calculated by a quantitative microbiological risk assessment (QMRA) approach. A modular process risk model from raw cocoa beans to cocoa liquor up to a hypothetical final product (70-90% dark chocolate tablet), was set up to understand changes of Salmonella concentrations following the production process. Different thermal treatments during bean or nib steam, nib roasting or liquor sterilization (achieving a 0-6 log reduction of Salmonella) were simulated. Based on the generic FAO/WHO Salmonella dose-response model and the chocolate consumption data in Belgium, salmonellosis risk per serving and cases per year at population level were estimated. When a 5 log reduction of Salmonella was achieved, the estimated mean risk per serving was 3.35×10-8 (95% CI: 3.27×10-10-1.59×10-7), and estimated salmonellosis cases per year (11.7 million population) was 88 (95% CI: <1-418). The estimated mean risk per serving was 3.35×10-9 (95% CI: 3.27×10-11-1.59×10-8), and the estimated salmonellosis cases per year was 9 (95% CI: <1-42), for a 6 log reduction. The current QMRA model solely considered Salmonella reduction in a single-step thermal treatment in the cocoa process. Inactivation obtained during other process steps (e.g. grinding) might occur but was not considered. As the purpose was to use QMRA as a tool to evaluate the log reduction in the cocoa processing, no post-contamination from the processing environment and ingredients was included. A minimum of 5 log reduction of Salmonella in the single-step thermal treatment of cocoa process, was considered to be adequate.

Optimization of HS-SPME/GC-MS Method for Determining Volatile Organic Compounds and Sensory Profile in Cocoa Honey from Different Cocoa Varieties (Theobroma cacao L.).

This study aimed to develop an analytical method using HS-SPME/GC-MS to determine the volatile organic compound (VOC) profiles and evaluate the sensory attributes of cocoa honey from four cocoa varieties (CCN51, PS1319, SJ02, and Parazinho). Using a multivariate factorial experimental design, the HS-SPME/GC-MS method was optimized to determine the VOC profiles. Twenty previously trained tasters participated in the ranking descriptive analysis, while 108 consumers participated in the acceptance and purchase intention tests. A total of 84 volatile organic compounds were identified from various chemical classes, including acids, alcohols, aldehydes, esters, ketones, monoterpenes, oxygenated monoterpenoids, sesquiterpenes, and oxygenated sesquiterpenoids. Palmitic acid was the compound found in the highest concentration in all varieties (5.13-13.10%). Multivariate analysis tools identified key compounds for differentiation and grouping of the samples. The results revealed that the variety significantly influenced both the VOCs' concentrations and sensory profiles. The CCN51, PS1319, and SJ02 varieties exhibited the highest diversity of VOCs and sensory attributes. Notably, the SJ02 and CCN51 varieties demonstrated superior acceptability and purchase intention, with means ranging from 7.21 and 7.08 to 3.71 and 3.56, respectively. These results indicate their potential as promising sources of cocoa honey for the food industry.

Growing cocoa in semi-arid climate and the rhythmicity of stem growth and leaf flushing determined by dendrometers.

This study investigated the performance of cocoa trees within an irrigated cocoa plantation situated in the semi-arid region of Bahia, Brazil. Two treatments were compared: "full sun," where cocoa trees were not shaded, and "shade," where trees were covered with a shading net absorbing 30 % of the radiation. The number of leaves and the leaf area index (LAI) were assessed using destructive method on 8 trees. In addition, new flushing of leaves, categorized into four flushing stages, were assessed visually on a weekly basis during two years. The variation of the stem diameter was measured using dendrometer sensors (n = 12 trees). Yield parameters like dry bean yield and number of fruits (healthy and aborted) were assessed on 40 trees per treatment. Both treatments, performed well in the semi-arid region. Generative parameters, such as dry bean yield (±2,000 kg/ha), fruit healthy and abortion rate per plot, were unaffected by full sun and shade treatments. The treatments showed high fruit abortion rates of (±60 %), showing that there's still much room for yield optimization. Additionally, stem diameter of the trees showed a significant reduction of the stem growth (daily increase of stem diameter) and maximum daily shrinkage (daily variation of stem diameter) during the flushing of new leaves. This implies that the emergence of new leaves significantly influences stem growth, consequently affecting the fruits which are growing on the stem. This assumption was corroborated by the significantly increased fruit abortion rate during the flushing of new leaves (stages 1 & 2). These findings highlight the potential of dendrometers to quantify this effect what can be used in future to optimize management practices. By doing so, more effective strategies can be developed to enhance cocoa yield and overall productivity in semi-arid regions.

Cocoa and Polyphenol-Rich Cocoa Fractions Fail to Improve Acute Colonic Inflammation in Dextran Sulfate Sodium-Treated Mice.

SCOPE: A study is conducted to determine the anti-inflammatory effects of cocoa and polyphenol-rich cocoa fractions in the dextran sulfate sodium (DSS)-induced mouse model of acute colonic inflammation. METHODS AND RESULTS: Male C57BL/6J mice are treated with dietary cocoa powder, an extractable cocoa polyphenol fraction, or a non-extractable cocoa polyphenol fraction for 2 weeks prior to treatment with 2.5% DSS in the drinking water for 7 days to induce colonic inflammation. Cocoa treatment continues during the DSS period. Cocoa and/or cocoa fractions exacerbate DSS-induced weight loss and fail to mitigate DSS-induced colon shortening but do improve splenomegaly. Cocoa/cocoa fraction treatment fails to mitigate DSS-induced mRNA and protein markers of inflammation. Principal component analysis shows overlap between cocoa or cocoa fraction-treated mice and DSS-induced controls, but separation from mice not treated with DSS. CONCLUSION: The results suggest cocoa and cocoa polyphenols may not be useful in mitigating acute colonic inflammation.

Inclusion of cocoa bran in the diet of lambs and its effect on reproductive parameters.

The objective of the study was to evaluate the effect of the inclusion of cocoa bran in the diet of lambs and its effect on reproductive parameters. For this, 40 lambs were randomly assigned to four treatments, and including 0, 10, 20 and 30% levels of cocoa bran in the concentrate. Blood was collected to measure cholesterol and testosterone and semen for physical and morphological evaluation; testicular biometry and morphometry were also evaluated. There was significant difference (P < 0.05) in body weight and tubulosomatic index between the lambs in the control treatment and those in the 30% cocoa bran treatment. There was no difference in testicular biometry, physical and morphological parameters of fresh semen, testicular morphometry, and volumetric ratio between lambs in all the treatments (P < 0.05). In addition, there was no difference in plasma cholesterol or testosterone concentration (P > 0.05). Thus, it is possible to include up to 30% of cocoa bran in diet without affecting the reproductive parameters of lambs.

Allergy to Theobroma Cacao.

We present the case of a patient with antecedent of tree nuts allergy who developed an anaphylactic reaction after ingesting a piece chocolate. An allergy study detected sensitization to cocoa in skin tests as well as cross-reactivity with tree nuts in the SDS-PAGE immunoblotting-inhibition.

Preparation and evaluation of functional cocoa-free spread alternatives from different sources.

Spread products have an important market share as they have high nutritional value and they are increasingly consumed, especially by children as a source of energy. The purpose of this work was to evaluate the potential use of powdered chickpea, black rice, carob, doum, date seeds, and beetroot to produce novel functional spreadable products as cocoa-free alternatives. Additionally, to avoid the side effects of cocoa-based products and to assess the cocoa replacement effects on the sensory properties, chemical composition, texture analysis, viscosity, antioxidant, peroxide stability, and microbial quality during storage periods were compared to the ones of cocoa spread. Sensory evaluation revealed that most formulated spreads were accepted as chocolate spread alternatives since there was no significant difference in overall acceptability among cocoa, chickpea, black rice, carob, and doum, while date seeds and beetroot spreads were significantly less acceptable. A variation was observed in the proximate chemical analysis of the produced functional spreads, as the alternative spreads had different characteristics to each other in their physicochemical, texture, and rheological properties. Results indicated a wide variation in the total phenolic content (TPC) of the different spread extracts. The highest amount of TPC was obtained for beetroot spread (455.84 mg GAE/100 g) followed by black rice spread (436.08 mg GAE/100 g). The obtained results indicated that the antioxidant activity of different spreads was significantly different (p < .05) while based on their microbiological analysis, they could have a shelf life of up to 9 months. According to the results, chickpea, carob, doum, black rice, date seeds, and beetroot powders could be used for the production of cocoa-free alternatives as they were highly acceptable and they showed antioxidant and antimicrobial activity.

Effects of Cocoa Consumption on Cardiometabolic Risk Markers: Meta-Analysis of Randomized Controlled Trials.

BACKGROUND: We conducted a systematic review and meta-analysis to examine the effect of dietary intake of cocoa on anthropometric measurements, lipid and glycemic profiles, and blood pressure levels in adults, with and without comorbidities. METHODS: The databases used were MEDLINE (PubMed), EMBASE, Web of Science, Cochrane, LILACS, and SciELO. The eligible studies were randomized clinical trials (RCTs) involving adults undergoing cocoa consumption (cocoa extract or ≥70% cocoa dark chocolate) for ≥4 weeks that evaluated at least one of the following markers: body weight, body mass index (BMI), waist/abdominal circumference, total cholesterol, LDL-c, triglycerides, HDL-c, blood glucose, glycated hemoglobin (HbA1c), and systolic and diastolic blood pressure (SBP/DBP). RESULTS: Thirty-one studies were included, totaling 1986 participants. Cocoa consumption showed no effects on body weight, BMI, waist circumference, triglycerides, HDL-c and HbA1c. Yet, there was a reduction in total cholesterol (-8.35 mg/dL, 95% CI -14.01; -2.69 mg/dL), LDL-c (-9.47 mg/dL, 95% CI -13.75; -5.20 mg/dL), fasting blood glucose (-4.91 mg/dL, 95% CI -8.29; -1.52 mg/dL), SBP (-2.52 mmHg, 95% CI -4.17; -0.88 mmHg), and DBP (-1.58 mmHg, 95% CI -2.54; -0.62 mmHg). CONCLUSIONS: The consumption of cocoa showed protective effects on major cardiometabolic risk markers that have a clinical impact in terms of cardiovascular risk reduction.

Effect of dark chocolate/ cocoa consumption on oxidative stress and inflammation in adults: A GRADE-assessed systematic review and dose-response meta-analysis of controlled trials.

BACKGROUND: Oxidative stress and inflammation play critical roles in the pathogenesis of many chronic diseases. Dark chocolate (DC)/cocoa, as a rich source of polyphenols like flavonoids, has anti-inflammatory and antioxidant properties that may confer health benefits, but findings in this context are inconsistent. OBJECTIVE: This systematic review and dose-response meta-analysis aimed to provide a comprehensive overview of the controlled trials (CTs) that have examined the effects of DC/cocoa on oxidative stress and inflammation biomarkers in adults. SEARCH METHODS: Databases including PubMed, Web of Science, and Scopus, were searched for relevant studies through April 2024. SELECTION CRITERIA: Studies assessed C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), malondialdehyde (MDA), nitric oxide (NO), P-selectin, E-selectin and thiobarbituric acid reactive substances (TBARS) in adults were included. DATA ANALYSIS: Based on the random-effects model, we calculated WMDs, SMDs and 95 % confidence intervals (CIs). Sensitivity, sub-group, meta-regression and dose-response analyses were also conducted. RESULTS: Thirty-three eligible CTs with 1379 participants were included. All studies reported the intervention types (cocoa powder, beverages and chocolate bars) and dosage. However, sixteen studies didn't do/report testing for purity and potency by independent groups. Also, none of the studies mentioned the risk of contamination with heavy metals. Another limitation was the lack of blinding assessment in studies. DC/cocoa significantly reduced MDA (SMD: -0.69, 95 %CI: -1.17, -0.2, p = 0.005) and increased NO levels (SMD: 2.43, 95 %CI: 1.11,3.75, p < 0.001); However, it has no significant effects on the other outcomes. Greater anti-inflammatory effects occurred at higher flavonoid doses (>450 mg/day) and for shorter durations (≤4 weeks) in the non-healthy participants. Non-linear dose-response relationships between cocoa dosage and CRP level and also between flavonoid dosage and IL-6 level were observed. Based on the GRADE evaluation, just CRP and MDA results were considered as high certainty evidence and the other outcomes results were categorized as very low to moderate certainty. CONCLUSIONS: DC/cocoa may improve systemic oxidative status and inflammation in adults. However, further studies should be performed to determine its benefits.

Characterization and Implementation of Cocoa Pod Husk as a Reinforcing Agent to Obtain Thermoplastic Starches and Bio-Based Composite Materials.

When the cocoa pod husk (CPH) is used and processed, two types of flour were obtained and can be differentiated by particle size, fine flour (FFCH), and coarse flour (CFCH) and can be used as a possible reinforcement for the development of bio-based composite materials. Each flour was obtained from chopping, drying by forced convection, milling by blades, and sieving using the 100 mesh/bottom according to the Tyler series. Their physicochemical, thermal, and structural characterization made it possible to identify the lower presence of lignin and higher proportions of cellulose and pectin in FFCH. Based on the properties identified in FFCH, it was included in the processing of thermoplastic starch (TPS) from the plantain pulp (Musa paradisiaca) and its respective bio-based composite material using plantain peel short fiber (PPSF) as a reinforcing agent using the following sequence of processing techniques: extrusion, internal mixing, and compression molding. The influence of FFCH contributed to the increase in ultimate tensile strength (7.59 MPa) and higher matrix-reinforcement interaction when obtaining the freshly processed composite material (day 0) when compared to the bio-based composite material with higher FCP content (30%) in the absence of FFCH. As for the disadvantages of FFCH, reduced thermal stability (323.57 to 300.47 °C) and losses in ultimate tensile strength (0.73 MPa) and modulus of elasticity (142.53 to 26.17 MPa) during storage progress were identified. In the case of TPS, the strengthening action of FFCH was not evident. Finally, the use of CFCH was not considered for the elaboration of the bio-based composite material because it reached a higher lignin content than FFCH, which was expected to decrease its affinity with the TPS matrix, resulting in lower mechanical properties in the material.

Microbiological characterization of kombucha and biocellulose film produced with black tea and cocoa bean shell infusion.

The food industry is increasingly striving to produce probiotics-based food and beverages using sustainable processes. Therefore, the use of by-products in product development has been investigated by several authors. The aim of this work was to investigate the effects of cocoa bean shell infusion in the production of kombucha through microbiological and genetic characterization. Three beverage formulations were prepared, one based on black tea (KBT), one based on cocoa bean shell infusion (KCS) and one containing 50 % black tea and 50 % cocoa shell infusion (KBL). The infusions were prepared with water, filtered, and sucrose was added. They were then homogenized and a portion of finished kombucha and SCOBY (symbiotic culture of bacteria and yeast) were added. Fermentation took place for 13 days and aliquots were collected every three days for physicochemical and microbial count analyses. Samples from the last day of fermentation were sent for DNA sequencing, extraction and quantification. The results were subjected to analysis of variance and compared by using Tukey's test (p < 0.05). The results show that there was a significant decrease in pH over time in all samples, while the titratable acidity increased, indicating an acidification of the beverage due to the production of organic acids. There was an increase in lactic acid bacterial colonies in all the formulations, which have a probiotic nature and are not always found in this type of beverage. Regarding the taxonomic classification of the samples, microorganisms of the kingdoms Fungi and Bacteria, of the families Saccharomycetaceae and Acetobacteraceae, were found in KBT, KCS and KBL, but with different microbiological compositions, with different amounts of yeasts and bacteria. Therefore, the use of by-products such as cocoa bean shell in the production of kombucha can contribute to the reduction of waste in the food industry and, at the same time, accelerate fermentation increasing the presence of lactic acid bacteria when compared to black tea.

Gut Dysbiosis Shaped by Cocoa Butter-Based Sucrose-Free HFD Leads to Steatohepatitis, and Insulin Resistance in Mice.

BACKGROUND: High-fat diets cause gut dysbiosis and promote triglyceride accumulation, obesity, gut permeability changes, inflammation, and insulin resistance. Both cocoa butter and fish oil are considered to be a part of healthy diets. However, their differential effects on gut microbiome perturbations in mice fed high concentrations of these fats, in the absence of sucrose, remains to be elucidated. The aim of the study was to test whether the sucrose-free cocoa butter-based high-fat diet (C-HFD) feeding in mice leads to gut dysbiosis that associates with a pathologic phenotype marked by hepatic steatosis, low-grade inflammation, perturbed glucose homeostasis, and insulin resistance, compared with control mice fed the fish oil based high-fat diet (F-HFD). RESULTS: C57BL/6 mice (5-6 mice/group) were fed two types of high fat diets (C-HFD and F-HFD) for 24 weeks. No significant difference was found in the liver weight or total body weight between the two groups. The 16S rRNA sequencing of gut bacterial samples displayed gut dysbiosis in C-HFD group, with differentially-altered microbial diversity or relative abundances. Bacteroidetes, Firmicutes, and Proteobacteria were highly abundant in C-HFD group, while the Verrucomicrobia, Saccharibacteria (TM7), Actinobacteria, and Tenericutes were more abundant in F-HFD group. Other taxa in C-HFD group included the Bacteroides, Odoribacter, Sutterella, Firmicutes bacterium (AF12), Anaeroplasma, Roseburia, and Parabacteroides distasonis. An increased Firmicutes/Bacteroidetes (F/B) ratio in C-HFD group, compared with F-HFD group, indicated the gut dysbiosis. These gut bacterial changes in C-HFD group had predicted associations with fatty liver disease and with lipogenic, inflammatory, glucose metabolic, and insulin signaling pathways. Consistent with its microbiome shift, the C-HFD group showed hepatic inflammation and steatosis, high fasting blood glucose, insulin resistance, increased hepatic de novo lipogenesis (Acetyl CoA carboxylases 1 (Acaca), Fatty acid synthase (Fasn), Stearoyl-CoA desaturase-1 (Scd1), Elongation of long-chain fatty acids family member 6 (Elovl6), Peroxisome proliferator-activated receptor-gamma (Pparg) and cholesterol synthesis (ß-(hydroxy ß-methylglutaryl-CoA reductase (Hmgcr). Non-significant differences were observed regarding fatty acid uptake (Cluster of differentiation 36 (CD36), Fatty acid binding protein-1 (Fabp1) and efflux (ATP-binding cassette G1 (Abcg1), Microsomal TG transfer protein (Mttp) in C-HFD group, compared with F-HFD group. The C-HFD group also displayed increased gene expression of inflammatory markers including Tumor necrosis factor alpha (Tnfa), C-C motif chemokine ligand 2 (Ccl2), and Interleukin-12 (Il12), as well as a tendency for liver fibrosis. CONCLUSION: These findings suggest that the sucrose-free C-HFD feeding in mice induces gut dysbiosis which associates with liver inflammation, steatosis, glucose intolerance and insulin resistance.

Chromatographic and spectroscopic methods for the detection of cocoa butter in cocoa and its derivatives: A review.

Currently, there is fierce competition in the cocoa industry to develop products that possess distinctive sensory characteristics and flavours. This is because cocoa and its derivatives provide numerous health and functional advantages, which is essential to their economics. The fatty acid and triglyceride composition of cocoa determines its quality. This review emphasises the necessity of developing precise, adaptable analytical techniques to identify and quantify cocoa butter in cocoa and its derived products, from cocoa beans to chocolate bars. Key chromatographic and spectroscopic techniques play crucial roles in understanding the fundamental principles underlying the production of cocoa with desirable flavours. This significantly impacts the sustainability, traceability, and authenticity of cocoa products while also supporting the battle against adulteration.

Fabrication and characterization of cocoa butter-based caffeine fast-melting tablets.

Aim: The objective of this study was to develop and characterize the physical properties of fast-melting tablets (FMTs) using cocoa butter as the base and caffeine as the model drug. Method: The simple refrigerator freezing method was employed to prepare caffeine-loaded, FMTs from cocoa butter bases. Results: The F3 chosen formulation achieved a disintegration time of 1.20 min ± 0.035, which falls within the specified limit set by the European Pharmacopoeia. The cumulative drug release data of F3, was 88.52 and 94.08% within 60 and 75 min, respectively (NLT 85% as per US FDA requirement). All the other physical test standards for FMTs met the pharmacopeial specifications. Conclusion: Based on the findings, the simple refrigerator freezing method could be used to formulate FMTs.

Patient-friendly natural caffeine-loaded cocoa butter-based fast-melting tablets with rapid disintegration, affordability, safety and biocompatibility are an efficient base for drug delivery.

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.

Assessment of pesticide knowledge, Safety Practices and postharvest handling among cocoa farmers in South Western Nigeria.

Cocoa beans are susceptible to fungal contamination during processing and storage. The knowledge of the use of pesticides and post-harvest handling of cocoa beans among farmers is of great importance for safe consumption. The study evaluated common cocoa production and post-harvest practices of farmers in selected study locations in South Western Nigeria. Primary data were collected through the administration of structured questionnaires, and interviews. The collected data were analyzed using inferential descriptive statistics. The results of 394 farmers showed that 52.9 % in Osun and 47.3 % in Oyo were primarily farmers by occupation, the rest had other ventures. The majority of cocoa farmers were men:83.6 % in Oyo State, 88.2 % in Osun state and 87.9 % in Ondo state. 28.6 % and 32.7 % of farmers were aged 51-60 in the Ondo and Oyo communities, respectively. Osun farming communities are dominated by young adults (51 %) of 31-50 years, followed by Ondo 40 % and36 % of farmers in Oyo State. Most cocoa farmers were married with 4-6 children as the most common household size in Osun (51 %), Ondo (60.4 %) and 49.1 % in Oyo State. The literacy level of farmers in cocoa communities was the highest in Oyo state where 47.3 % had tertiary education. Farmers in Oyo State had better knowledge of the dangers of pesticides than Ondo and Osun. However, ignorance of dangers in agrochemicals was higher among Osun farmers than in Ondo State. The highest (18 %) pesticide use during storage was recorded among Oyo farmers, while the least (11.0 %) was recorded among farmers in Ondo State. Pesticide usage was more abundant in Osun (50 %) during cocoa production than in the other study areas. The majority of farmers were positively disposed to make use of nose masks during agrochemical application, meanwhile, 69 %, 62 %, and 61 % of farmers used them already in Oyo, Ondo, and Osun states, respectively. Educational qualification (χ2 = 9.176, p = 0.027) of cocoa farmers was significantly related to knowledge of best practices. Farmers with higher education have a greater ability to receive and process information relating to global best practices in production, postharvest, and pesticide handling in cocoa. In conclusion, cocoa farmers' knowledge of processing, use of pesticides, and storage practices differed from one location to another. Intensive orientation and more enlightenment by extension workers against indiscriminate use of pesticides in cocoa plantations and stores must be consistently and continuously done.

Production of low-protein cocoa powder with enzyme-assisted hydrolysis.

Amino acid-related disorders are caused by a defect in the metabolic pathways of amino acid groups. These patients must follow a lifelong protein diet. The objective of this study was to produce a low-protein cocoa powder (LPP) with enzymatic hydrolysis and precipitation method. First, the solubility of cocoa powder was increased by heat and enzyme treatments (Amylase, Viscozyme, and Alcalase). Then, the protein level was decreased by isoelectric precipitation. According to the obtained results, the solubility of cocoa powder rose from 28.61% to 50.69%. Protein content decreased by almost 40% and significant reductions in the amino acid profile were also provided; the highest ones were detected in methionine (100%), lysine (73.65%), leucine (53.64%), alanine (46.17%), and isoleucine (44.73%) levels. LPP had high phenolic content (25.10 mg/g GAE) and the changes in the antioxidant activities were not significant (p > .05). Moreover, chocolate production with LPP and control powder was also carried out under laboratory conditions. Hardness (1732.52 g), moisture content (0.60%), and water activity (0.37) of chocolate samples produced with low-protein cocoa powder (LPC) were lower than those of the control sample. The Casson model well fitted to the rheological data (R 2 > .990) and chocolate samples showed elastic behavior. The removal of proteins from the cocoa was verified with Fourier transform infrared spectroscopy analyses. The melting temperatures of chocolates (31.84 and 31.54°C for control and LPC samples, respectively) did not change with the applied process. As a conclusion, it was revealed that the production of low-protein cocoa powder and chocolate is feasible for patients with amino acid disorders with this study.