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In this study, we developed and validated a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of eight phytocannabinoids in various cannabidiol (CBD) products from Japanese market. This method was combined with electrospray ionization in positive mode and sample preparation with QuEChERS. Three types of commercial products such as honey, chocolate, and gummies were used to perform accurate quantification with unified protocol of LC-MS/MS and QuEChERS. The limit of detection and quantification were 5-20 µg g -1 and 10-40 µg g -1 , respectively. Reproducibility was ensured using matrices free of target foods, resulting in an accuracy within ±10 % and a precision with a relative standard deviation of less than 5 % for all targets. Finally, this analytical method was applied to 8 series of commercial samples from the Japanese market. This unified protocol will serve as a reference as an official method in Japan., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Cocoa (Theobroma cacao L.) is the basic raw material to produce chocolate and other derivatives such as cocoa butter, cocoa powder and cocoa liquor (cocoa paste), which requires a fermentation process that affects its chemical composition and sensory profile. The objective of this study was to monitor the biochemical, physical and sensory changes during fermentation of cocoa beans in cocoa bean processing plants in the department of Caquetá, Colombia. During fermentation, the temperature of the mass and the pH of the pulp and beans were monitored at the different cocoa bean processing plants (Sites ASOACASAN ASA, COMICACAO CMI, COMCAP COC). Also, at two points during fermentation (days 4 and 7), physical properties of the bean were determined, such as variables related to bromatological composition, polyphenolic compounds and antioxidant activity as sensory attributes at the different sites. An increase in dough temperature was found, however the pH of the cotyledon decreased during the fermentation process and the fat and moisture content varied with fermentation time. At the site level, total polyphenol content (TPC), total flavonoids (TF), 1,1-diphenyl-2-picrylhydrazil (DPPH) and ferric reducing antioxidant power (FRAP) contents were statistically different, with COC being different from the other sites. The TPC was higher at the COC site (507 mg gallic acid equivalent GAE/g Cocoa) with respect to the other sites (< 360 mg GAE/g Cocoa). The TF content followed a similar behavior to TPC, with significant differences between sites and differences between fermentation times for ASA. The TF was higher in COC (309.1 mg catechin/g cocoa) with respect to CMI (215.6 mg catechin/g cocoa) and ASA (185.7 mg catechin/g cocoa). Values in DPPH ranged from 5869.3 to 7781.8 μmol Trolox/g cocoa and for the FRAP assay ranged from 369.8 to 606.7 mg ascorbic acid AA/g cocoa among the sites. It was found that the time and management of the fermentation process has a significant impact on the parameters (biochemical, physical and sensory) of cocoa beans. Therefore, it is necessary to standardize the fermentation process to achieve a quality product that meets the needs of the market., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 González et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Dark chocolate produced on equipment used to manufacture milk chocolate can contain milk due to cross-contact. This study evaluated the use of dry cleaning methods for removing milk chocolate residue from a butterfly or ball valve attached to a stainless steel pipe and from pilot-scale equipment used in chocolate manufacture. Milk-free dark chocolate (40 °C) was pumped through a milk chocolate-contaminated valve/pipe assembly after no cleaning, use of a pig purging treatment, or a 40 °C cocoa butter flush. Dark chocolate samples were collected at 7-sec intervals. Treatments investigated for the removal of residual milk chocolate from a conche and a ball mill included no cleaning, a 40 °C cocoa butter rinse, and wet cleaning. After cleaning, three batches of dark chocolate (40 °C) were processed in the ball mill and conche, and each batch was collected. Milk chocolate was processed on a 3-roll refiner, followed by push-through with dark chocolate (∼9 kg) with 0.3 kg samples collected at 5-min intervals. Dark chocolate samples were analyzed for milk concentrations by ELISA. Trials and analyses were completed in triplicate. Dark chocolate push-through alone resulted in milk concentrations ≥4,500 µg/g in samples obtained from the contaminated valve/pipe combinations within the first few seconds of collection, and ≥16.2 kg of dark chocolate was needed to obtain milk concentrations below the ELISA LOQ (2.5 µg/g). A pig purging treatment of the ball valve/pipe assembly resulted in milk concentrations below the ELISA LOQ. A cocoa butter flush of the butterfly valve/pipe decreased initial milk concentrations, but milk was detected until ≥18.7 kg dark chocolate purge. Milk concentrations in first batches of dark chocolate processed in a ball mill and conche without cleaning were ≥17,000 µg/g while the use of a cocoa butter rinse reduced milk levels in dark chocolate by ≥89%. Some dry cleaning treatments were effective at reducing levels of milk in dark chocolate due to cross-contact., (Published by Elsevier Inc.)

This study investigated the use of microencapsulated Asiatic pennywort (Centella asiatica) (CA) as a functional ingredient to formulate a novel chocolate oat milk beverage. The main objectives of the study were to characterize and encapsulate bioactive components from CA and to determine the polyphenol content and sensory properties of the beverage. CA extract was microencapsulated using maltodextrin and gum Arabic as carriers and subsequently freeze-dried to produce microcapsules. Microencapsulated CA was incorporated into chocolate oat milk at varying concentrations. Polyphenol content of the beverages was quantified using liquid chromatography-mass spectrometry. Consumer acceptability and sensory perception of the beverages were evaluated through an acceptance test and a check-all-that-apply test, respectively, to assess the sensory characteristics of the chocolate oat milk beverage. CA fortified chocolate oat milk contained fourteen polyphenols. Increasing the concentration of microencapsulated CA led to an increase in the polyphenol content of the beverage. Among the identified polyphenols, asiatic acid and asiaticoside stood out as the unique and most abundant compounds in CA (p < 0.05). Additionally, the incorporation of cocoa powder into the beverage further contributed to the polyphenol content, introducing bioactive compounds such as benzoic acid, caffeic acid, catechin, chlorogenic acid, kaempferol, luteolin, madecassic acid, p-coumaric acid, and quercetin. Evaluation of consumer acceptability revealed that chocolate oat milk beverages containing 2% and 4% microencapsulated CA were liked by consumers. However, beverages with higher concentrations of CA were perceived as less acceptable, characterized by grassy, bitter, and earthy attributes. In conclusion, this study demonstrates the potential of microencapsulated CA as a functional ingredient in chocolate oat milk beverages. PRACTICAL APPLICATION: This study reveals new insights on the microencapsulation of bioactive compounds in CA, proposing its potential as a novel functional ingredient in food and beverage applications in Western markets. The study revealed microencapsulated CA retained polyphenols in CA including asiatic acid and asiaticoside responsible for its bioactive properties. Consumer perception of CA added to oat milk revealed that it can be added at an acceptable level of 4%; however, higher amounts can decrease consumer acceptability. As practitioners explore the incorporation of CA as a functional component in food products, it is crucial to explore preservation techniques for the sensitive bioactive components while balancing the optimal amount of CA to enhance overall consumer liking., (© 2024 The Author(s). Journal of Food Science published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Background: In this study, innovative chocolate, citrus and mixture flavoured tofu-based nutritionally customised, dysphagia-oriented, comfortably consumed, appetising, one-bite-sized finger foods, oriented to malnutrition, sarcopenia and frailty prevention in older people were created by using 3D printing technology. Developed products were characterised by evaluating chemical composition and physical properties and performing sensory evaluation among geriatric clinic residents (≥60 years)., Results: The dietary composition of the developed foods was: 19-21 g (100 g) -1 protein, 6-8 g (100 g) -1 fibre, 8-9 g (100 g) -1 fat, 11 mg (100 g) -1 iron, 14 mg (100 g) -1 zinc, 70 μg (100 g) -1 selenium. Foods were also enriched with branched-chain amino acids, such as leucine, isoleucine and valine. All formulated foods were classified as level 6 by International Dysphagia Diet Standardisation Initiative classification. Chocolate-flavoured food was much harder (4914 g) with lower adhesiveness value (-33.6 g s), compared to the citrus- or mixture-flavoured foods. Older people evaluated all finger foods as very easy handled by hand, soft, easy to swallow, having a moderate flavour intensity and a weak afterfeel. Despite the fact that the chocolate food was evaluated as having the highest hardness and gumminess values by the instrumental method, this difference was not noticeable to the evaluators. However 7% of the participants said that 3D printed foods were sticky to dentures., Conclusion: The results suggest that it is possible to create nutrient-dense comfortably consumed 3D printed foods, oriented to malnutrition, sarcopenia and frailty prevention in older people. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

The objective of this study was to assess the effects of simulated digestion on the formation of α-dicarbonyl compounds (α-DCs) in chocolates. For that purpose, the concentrations of glyoxal and methylglyoxal in chocolates were determined through High-Performance Liquid Chromatography (HPLC) analysis before and after in vitro digestion. The initial concentrations ranged from 0.0 and 228.2 µg/100 g, and 0.0 and 555.1 for glyoxal and methylglyoxal, respectively. Following digestion, there was a significant increase in both glyoxal and methylglyoxal levels, reaching up to 1804 % and 859 %, respectively. The findings indicate that digestive system conditions facilitate the formation of advanced glycation end product (AGE) precursors. Also, glyoxal and methylglyoxal levels were found to be low in chocolate samples containing dark chocolate. In contrast, they were found to be high in samples containing hazelnuts, almonds, pistache, and milk. Further studies should focus on α-DCs formation under digestive system conditions, including the colon, to determine the effects of gut microbiota., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Roasted carob pulp (Ceratonia siliqua) is a cocoa substitute known for its faint cocoa-like resemblance. However, the cocoa-carob flavour gap remains poorly uncharacterised. This study aimed to elucidate the sensory and molecular aspects of this flavour gap in a 70 % dark chocolate formulation via a two-pronged instrumental-sensorial approach. Descriptive Sensory Analysis (DSA) revealed carob-based chocolate was significantly sweeter, less sour and astringent than conventional dark chocolate due to the high total sugar content (45-50 % DM; HPLC/RID), low titratable acidity and tannin content, respectively. As for aroma, a distinct, albeit weak, cocoa-like aroma was present in carob-based chocolate. HS-SPME-GC-MS/FID revealed this was attributed to branched-chain Strecker aldehyde generation during roasting (2-methylbutanal, 1.17 μg/g; 3-methylbutanal, 2.89 μg/g). Notably, there was a distinct lack of alkylpyrazines. Additionally, a distinct woody, tree bark-like odour was uniquely associated with carob-based chocolates. This was due to furfural generation during roasting (2.33 μg/g). In conclusion, the aroma and taste gap between cocoa and carob was successfully characterised in this study. These findings substantiate the potential of carob application in chocolate manufacturing, thus empowering confectioners to make evidence-based decisions when evaluating cocoa substitutes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The presence of contaminants in cacao-derived products, especially in chocolates, has raised concerns regarding food safety and human health. The study assessed the concentration variation of 16 elements in 155 chocolate samples from the US market by cacao content and country of geographic origin. The study further examined the potential health risks posed by toxic metals and determined the contribution of essential elements to the Daily Recommended Intake (DRI), estimated based on an ounce (∼28.4 g) of daily chocolate consumption. Dark chocolates with ≥50 % cacao exhibited consecutively increasing mean levels from 1.2 to 391 µg/kg for U, Tl, Th, As, Pb, Se, Cd, and Co. Similarly, Ni, Sr, Cu, Mn, Zn, Fe, Ca, and Mg had mean concentrations from 4.0 to 1890 mg/kg. Dark chocolates sourced from Central and South America exhibited the highest mean levels of Cd, and South America samples also contained elevated Pb, whereas those from West Africa and Asia had low Cd and Pb, respectively. Cacao contents showed increasingly strong association with Cd, Co, Mn, Sr, Ni, Cu, Zn, and Mg (r = 0.60-0.84), and moderately with Se, Fe, As, and Tl (r = 0.35-0.49), indicating these elements are primarily derived from cacao beans. Weak association of cacao contents with Pb, Th, and U levels (r < 0.25), indicates post-harvest contaminations. Hazard Quotient (HQ) > 1 was found only for Cd in 4 dark chocolates, and Hazard Index (HI) > 1 for cumulative risk of Cd, Pb, Ni, As, and U was found in 33 dark chocolates, indicating potential non-carcinogenic risks for 15 kg children but none for 70 kg adults. Dark chocolate also substantially contributed to 47-95 % of the DRI of Cu for children and 50 % for adults. Dark chocolates also provided notable Fe, Mn, Mg, and Zn contributions to the DRI. These essential elements are recognized to reduce the bioavailability of toxic metals such as Cd, Pb, or Ni, thereby potentially lowering associated health risks. This study informs consumers, food industries, and regulatory agencies to target cacao origins or chocolate brands with lower toxic metal contents for food safety and minimizing adverse health effects., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Among the emerging prebiotics, galactooligosaccharide (GOS) has a remarkable value with health-promoting properties confirmed by several studies. In addition, the application of ohmic heating has been gaining prominence in food processing, due to its various technological and nutritional benefits. This study focuses on the transformative potential of ohmic heating processing (OH, voltage values 30 and 60 V, frequencies 100, 300, and 500 Hz, respectively) in prebiotic chocolate milk beverage (3.0 %w/v galactooligosaccharide) processing. Chemical stability of GOS was assessed along all the ohmic conditions. In addition, microbiological analysis (predictive modeling), physical analysis (color and rheology), thermal load indicators assessment, bioactivity values, and volatile compound was performed. HPAEC-PAD analysis confirmed GOS stability and volatile compound evaluation supported OH's ability to preserve flavor-associated compounds. Besides, OH treatments demonstrated superior microbial reduction and decreased thermal load indicators as well as the assessment of the bioactivity. In conclusion, OH presented was able to preserve the GOS chemical stability on chocolate milk beverages processing with positive effects of the intrinsic quality parameters of the product., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: The high quality and unique flavor and aroma of bean-to-bar chocolates have resulted in an increase in the consumption of these products. Nevertheless, cocoa beans may present inorganic contaminants from environmental and anthropogenic sources which can contribute to contamination of the chocolates, despite the fewer processing steps and few ingredients used in bean-to-bar manufacturing process compared to the industrial one. Therefore, this study aimed to evaluate the content of trace elements (As, Cd, Co, Cu, Hg, Pb, Se) in bean-to-bar chocolates and traceable cocoa beans from Brazil and Ecuador., Methods: Bean-to-bar chocolate samples were acquired in Brazil (n=65) and Ecuador (n=10), considering the main products available: white, milk, semisweet and dark chocolate. Cocoa samples from dedicated farms (n=23) were analyzed for trace elements and inorganic contaminants regulated by Brazil and European agencies. Samples were mineralized using acid digestion (nitric acid and hydrogen peroxide) in a closed microwave-assisted system. Quantification of trace elements was performed using Inductively coupled plasma mass spectroscopy (ICP-MS) and Inductively coupled plasma optical emission spectroscopy (ICP OES) in optimized conditions. The analytical control was performed with certified reference materials (ERM BD512 - Dark Chocolate, Tort-2 and Tort-3 - Lobster Hepatopancreas and SRM 1547 - Peach leaves) and recoveries ranged between 84% and 105% for all elements., Results: The trace element levels in the bean-to-bar chocolates were (mg/kg): As (<0.022-0.023), Cd (<0.002-0.74), Cu (0.11-21.2), Co (<0.003-1.88), Hg (<0.010-<0.010), Pb (<0.007-0.22), and Se (<0.029-0.35). The exposure assessment from inorganic contaminants in chocolates revealed up to 93% of provisional tolerable monthly intake (PTMI) for Cd and 123% of tolerable upper intake level (UL) for Co for children. Inorganic contaminants were also analyzed in cocoa beans from dedicated farms and Cd and Pb levels were found above the thresholds established by Brazil health agency., Conclusion: The results observed for both bean-to-bar chocolates and raw materials (cocoa beans from dedicated farms) indicated a need for monitoring these trace elements., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

The objective of this research was to evaluate changes in flow behavior of chocolate during chocolate grinding using a stone grinder as affected by chocolate formulation. Three different types of chocolates were evaluated. Two chocolates without milk added (70% chocolate) and two chocolates with milk added and with different amounts of cocoa nibs (30% chocolate and 14% chocolate) were tested. For the 70% chocolates, nibs of two different origins were used; therefore, a total of four samples were evaluated. Chocolates were processed in a stone grinder, and samples were taken as a function of grinding time. For each timepoint, the flow behavior of the samples was measured using a rotational rheometer and fitted to the Casson model. Particle size was measured using a laser scattering instrument. Results showed that yield stress increased linearly while the Casson plastic viscosity decreased exponentially with grinding time (smaller particles). Particle size distribution of the chocolates showed a prominent bimodal distribution for short grinding times (∼9 h) with small (∼15 µm) and large (∼100 µm) particles; with longer grinding time, the population of larger particles decreased. Yield stress values were higher for the 70% chocolate, but they were not very different between the two milk chocolates tested. The Casson plastic viscosity was greatest for the 70% chocolate, followed by the 30% chocolate. The 14% chocolate had the lowest Casson plastic viscosity. Changes of Casson plastic viscosity with particle size were more evident for the dark chocolates compared to the milk ones. These results are helpful to small chocolate producers who need better understanding of how the formulation and grinding of chocolate affect its flow behavior, which will ultimately affect chocolate handling during production., (© 2024 Institute of Food Technologists.)

The most significant contributor to global mortality are cardiovascular diseases. Dietary factors significantly impact the risk, advancement, and treatment of cardiometabolic conditions. Chocolate, known for its adaptability and capacity to stimulate pleasure centers, emerges as a promising vehicle for integrating different bioactive elements. This systematic review analyzed 10 randomized controlled trials investigating the health effects of consuming enriched, fortified, or supplemented chocolate. These trials varied in chocolate intake amounts (ranging from 5 to 101 g day -1 ), incorporated bioactive components (co-crystalized astaxanthin, lycopene, wood-based phytosterol-phytostanol mixture, canola sterol esters, etc .), and duration (from 2 weeks to 1 year). Some enriched chocolates were found to reduce total and LDL cholesterol and influence markers of oxidative damage, inflammation, immune function, and skin parameters. However, certain trials showed a minimal impact on health outcomes. Therefore, while enriched chocolate holds promise as a carrier for beneficial bioactive compounds, rigorous scientific inquiry and methodological rigor are crucial to fully substantiate these claims. Comprehensive evaluations covering cardiovascular health, metabolic function, immune response, and other aspects are needed to understand its potential benefits and limitations. Advancing robust research initiatives could help realize the full potential of enriched chocolate in promoting human health and well-being.

Background: Titanium dioxide (TiO 2 ) is banned in some countries but its use is still permitted in others. The global food supply chain is therefore challenged with the need to use rapid and reliable testing methods to either detect the presence of TiO 2 or to quantify its concentration. The goal of this study was to determine the feasibility of using color, texture profile analysis, Raman microscopy, and X-ray fluorescence (XRF) spectroscopy to detect and quantify TiO 2 in fillings used in the pastry and confectionery industry. In this study, two types of fillings were investigated: vanilla based and chocolate based. All fillings were prepared in four different variations - without TiO 2 and with three concentrations as follows: 0.25 g*kg -1 , 0.5 g*kg -1 , or 0.75 g*kg -1 TiO 2 per sample. The methods were selected for their ability to analyze the samples in a short period of time., Results: All of the methods showed moderate to high potential for detecting TiO 2 in the samples. The results reveal how TiO 2 affects the food matrix color and texture. Use of Raman microscopy confirms its detectability, although concentrations of TiO 2 do not follow a pattern. X-ray fluorescence spectroscopy showed the greatest potential as it can not only detect TiO 2 but can also quantify its concentration in the samples., Conclusions: The highest potential for quantifying the concentration of this food additive was achieved with XRF. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

The aim of this study was to produce chocolate using roasted (RB) and unroasted cocoa beans (URB). The effect of roasting on the total phenolic content (TPC), antioxidant activity [2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity, ferric reducing antioxidant power (FRAP), and cupric ion reducing antioxidant capacity (CUPRAC) values], phenolic compounds, caffeine, oxidative stability [free fatty acid, peroxide, conjugated dienes, conjugated trienes, and thiobarbituric acid reactive substances (TBARS)], Fourier transform infrared (FTIR), and differential scanning colorimetry (DSC) analysis of both cocoa beans and chocolate samples were analyzed. According to the results, the TPC of URB (24.96 mg gallic acid equivalent (GAE)/g sample) was higher than roasted beans (21.32 mg GAE/g sample). Similar results were also seen in the TPC of chocolate samples. Although roasting did not affect the DPPH scavenging activity and caffeine content of cocoa beans, it decreased FRAP and CUPRAC values. (-)-Epicatechin and chlorogenic acid values were higher in unroasted bean and chocolate samples, but the amount of gallic acid increased with the roasting process. Free fatty acid, peroxide, conjugated dienes, conjugated trienes, and TBARS results of unroasted samples were lower than roasted ones, indicating better oxidative stability. The melting temperatures of cocoa beans changed with roasting while it was similar between chocolate samples. Composition of the beans and the chocolate samples were qualitatively determined with FTIR., Competing Interests: DECLARATION OF CONFLICTING INTERESTSThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

To reduce unwanted fat bloom in the manufacturing and storage of chocolates, detailed knowledge of the chemical composition and molecular mobility of the oils and fats contained is required. Although the formation of fat bloom on chocolate products has been studied for many decades with regard to its prevention and reduction, questions on the molecular level still remain to be answered. Chocolate products with nut-based fillings are especially prone to undesirable fat bloom. The chemical composition of fat bloom is thought to be dominated by the triacylglycerides of the chocolate matrix, which migrate to the chocolate's surface and recrystallize there. Migration of oils from the fillings into the chocolate as driving force for fat bloom formation is an additional factor in the discussion. In this work, the migration was studied and confirmed by MRI, while the chemical composition of the fat bloom was measured by NMR spectroscopy and HPLC-MS, revealing the most important triacylglycerides in the fat bloom. The combination of HPLC-MS with NMR spectroscopy at 800 MHz allows for detailed chemical structure determination. A rapid routine was developed combining the two modalities, which was then applied to investigate the aging, the impact of chocolate composition, and the influence of hazelnut fillings processing parameters, such as the degree of roasting and grinding of the nuts or the mixing time, on fat bloom formation.

α,β-Unsaturated carbonyls are highly reactive and described as structural alerts for genotoxicity. Ten of them (either commercially available or synthesized here by combinatorial chemistry) were first investigated throughout the chocolate-making process by solvent-assisted flavor evaporation (SAFE) coupled to GC-MS/SIM. Monitored α,β-unsaturated aldehydes were formed during chocolate production, primarily through aldol condensation of Strecker aldehydes triggered by bean roasting. Notably, levels of 2-phenylbut-2-enal (up to 399 μg·kg -1 ) and 5-methyl-2-phenylhex-2-enal (up to 216 μg·kg -1 ) increased up to 40-fold. Dry conching caused evaporation of α,β-unsaturated carbonyls, while wet conching partially restored or increased their levels due to cocoa butter addition. Further analyses showed that α,β-unsaturated aldehydes also occurred in most commercial sweet snacks (up to 16 μg·kg -1 ), although often at lower concentrations than in roasted cocoa or derived chocolates. In the end, none of the monitored α,β-unsaturated aldehydes did raise a health concern compared to current maximum use levels (2-5 mg·kg -1 ). On the other hand, much higher levels of genotoxic furan-2( 5H )-one were found in crepe and cake samples (up to 4.3 mg·kg -1 ).

In the present study, the effect of the addition of quince and collagen type I and III to dessert chocolate on its functional properties was determined. The study evaluated the antioxidant potential of the tested formulations using the FRAP method and the linoleic acid oxidation test and beta-carotene bleaching test. The tested samples were also evaluated for inhibitory activity against enzymes important in preventive health (inflammation and neurodegenerative disorders) namely: AChE, BChE, GR, GPx, COX, and SOD. The addition of quince and collagen to the chocolate samples resulted in higher activity compared to the control sample, as indicated by the FRAP test. The experiment highlighted the impact of including quince fruit on the antioxidant activity of the chocolate samples. Interestingly, merely increasing the quince fruit amount did not consistently enhance antioxidant potential. Specifically, chocolate samples with a lower proportion of quince fruit (2 g/100 g) exhibited greater antioxidant activity when supplemented with collagen I. Conversely, in samples with higher quince percentages (3 g and 4 g), those enriched with collagen III showed higher antioxidant activity. Similar correlations were observed in the linoleic acid oxidation test. Notably, samples containing 3 g and 4 g of quince and type III collagen demonstrated statistically similar highest antioxidant properties. Regardless of the collagen type used, there was no observed increase in activity towards the tested enzymes for samples with the lowest percentage of quince fruit. Both collagen types exhibited the highest activity in the inhibition assay against acetylcholinesterase and butyrylcholinesterase when combined with 3 g and 4 g of quince. Overall, the experimental incorporation of both fruit and collagen enhanced the chocolates' activity. Similarly to the antioxidant activity findings, chocolates with lower quince fruit quantities showed increased activity when supplemented with collagen III, while those with higher quince content (3 g and 4 g) displayed higher activity with collagen I. Bitter chocolate by itself is an attractive food product, rich in many bioactive compounds. However, enriching it with other attractive raw materials can make its properties and taste even more attractive.

Background: Acrylamide (AA) is a process contaminant naturally formed during the cooking of starchy food at high temperatures. Considering existing risks of misquantification inherent to the analysis of AA, an AOAC initiative raised the need for a consensus standard to determine AA in a broad variety of food., Objective: A quantitative LC-MS/MS method for AA determination in food was validated in a single-laboratory study. Targeted performance requirements in terms of target matrixes, limit of quantification, recovery, and precision were as defined per Standard Method Performance Requirement (SMPR®) 2022.006., Method: The proposed method derives from EN 16618:2015 standard pending modifications brought to the (1) sample preparation (simplified, potentially automated); (2) scope of application (significantly extended); and (3) LC conditions (improved selectivity). Confirmatory detection of AA is conducted by LC-MS/MS in the Selected Reaction Monitoring mode (SRM), and isotopic dilution was applied for quantification approach using either 2,3,3-d3-acrylamide (d3-AA), or 13C3-2,3,3-d3-acrylamide (13C3-d3-AA) as labeled internal standard., Results: A total of 16 laboratory samples from nine matrix categories were included in the validation process. A full validation was conducted on coffee (instant, roast), infant cereal, cocoa powder, pet food (croquettes), tea (green tea), spices (black pepper), and nuts (roasted almonds) with satisfactory performances both in terms of recovery (97-108%) and precision (RSDr and RSDiR <12%). The method applicability was further demonstrated through the analysis of quality control materials and reference materials including French fries, potato crisps, vegetable crisps, instant coffee, infant food, and biscuits (cookies), with accuracy values determined within a 94-107% range., Conclusions: The performances of the presented method are in agreement with the acceptance criteria stipulated in SMPR 2022.006., Highlights: The Expert Review Panel for acrylamide approved the present method as AOAC Official First Action 2023.01., (© The Author(s) 2024. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

In this study, the effects of temperature (22, 24, 26, 28, and 30°C) and strain (0.1%, 1%, and 5%) on cocoa butter (CB) crystallization were investigated by oscillatory test, and the four-parameter Gompertz model was used to interpret the effect of parameters on pre-crystallization, nucleation, and crystal growth stages of CB. Lag time and growth rate were calculated using the Gompertz model using time-dependent storage modulus (G') data. According to the results, CB crystallization at 26°C with a 1% strain value had the highest growth rate value, the shortest lag time, and the formation of β v polymorph type. Followingly, polymorphic types of the CB crystals were determined based on the melting points of polymorphs via the temperature ramp step, and the results obtained were correlated with a polarized light microscope. In conclusion, using a rheometer in both the observation of the pre-crystallization process and the determination of polymorph types is very important for research and development studies in the chocolate industry for process and formulation optimization. PRACTICAL APPLICATION: This study demonstrates the feasibility of a novel approach for investigating crystallization and oscillatory shear of CB using a rheometer, both for observing crystallization kinetics and determining polymorph type, accompanied by the Gompertz equation to model the crystallization kinetics. According to the results, the effect of process parameters (temperature and shear) on the crystallization behavior of CB can be observed by rheometer, which can provide a detailed perspective for chocolate manufacturers and researchers in research and development studies., (© 2024 The Authors. Journal of Food Science published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Chocolate production faces nutritional, environmental and socio-economic challenges present in the conventional cocoa value chain. Here we developed an approach that addresses these challenges by repurposing the often-discarded pectin-rich cocoa pod endocarp and converting it into a gel. This is done using cocoa pulp juice concentrate to replace traditional sugar from sugar beets. Although swelling of fibres, proteins and starches can limit gel incorporation, our proposed chocolate formulation contains up to 20 wt% gel. It also has comparable sweet taste as traditional chocolate while offering improved nutritional value with higher fibre and reduced saturated fatty acid content. A cradle-to-factory life cycle assessment shows that large-scale production of this chocolate could reduce land use and global warming potential compared with average European dark chocolate production. The process also provides opportunities for diversification of farmers' income and technology transfer, offering potential socio-economic benefits for cocoa-producing regions., (© 2024. The Author(s).)

In this study, the effects of independent variables such as alkaline (NaOH) salt concentration (3.0-6.0 g/100 mL), alkalization temperature (60-90 °C), and time (20-40 min) on cocoa powder (low-fat) properties were investigated by using Central Composite Design. The physicochemical and color properties of samples, powder characteristics, volatile component profile, total polyphenol content (TPC), as well as antioxidant activity potentials using different methods (DPPH and ABTS) were determined. Significant models were identified for the effects on major alkalization indicators (L*, a*/b*, pH), as well as TPC and antioxidant activity potential (DPPH), which are the main motivators for the preference and consumption of cocoa products (p < 0.05). The established model was validated, and their predicted values were found to be very close to real results. It was determined that the alkali concentration had a more significant effect on dependent variables, especially on alkalization indicators, compared to the other independent variables. Furthermore, strong correlations were determined between TPC and antioxidant activity potential and color properties (L*, a*, b*, and a*/b*). Optimum concentration, temperature and time were found to be 5.3 %, 84 °C and 35.7 min for maximizing a*/b* value. The establishment of such models lead to optimizing process conditions of alkalization with minimum effort and labor force for obtaining cocoa powder with desired quality depending on the usage purpose., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Fine flavor cocoa (FFC) is known for its unique flavor and aroma characteristics, which vary by region. However, a comprehensive overview of the common sensory attributes used to describe FFC beans and chocolate is lacking. Therefore, a systematic review was conducted to analyze existing literature and identify the most commonly used sensory attributes to describe FFC beans and chocolate. A systematic search of the Web of Science and Scopus databases was conducted in May 2023, and Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed to ensure transparency and reproducibility. This review summarizes the origins of cocoa and explores their unique flavor profiles, encompassing caramel, fruity, floral, malty, nutty, and spicy notes. Although some origins may exhibit similar unique flavors, they are often described using more specific terms. Another main finding is that although differences in sensory attributes are anticipated at each production stage, discrepancies also arise between liquor and chocolate. Interestingly, fine chocolate as the final product does not consistently retain the distinctive flavors found in the liquor. These findings emphasize the need for precise descriptors in sensory evaluation to capture flavor profiles of each origin. As such, the exploration of attributes from bean to bar holds the potential to empower FFC farmers and chocolate producers to effectively maintain quality control., (© 2024 Institute of Food Technologists.)

Understanding mechanisms inside the conche is inevitable for targeted conching. Therefore, distribution and transition of aroma-active volatiles (acetic acid, benzaldehyde, (R,S)-(±)-linalool, 2,3,5,6-tetramethylpyrazine, 2-phenylethanol, and 2-phenylethyl acetate) between the different components of dark chocolate (cocoa butter, cocoa particles, sugar particles) were studied. Different model systems were designed and aroma-analytically analyzed by stable isotope dilution analysis. Diffusion mechanisms of selected aroma-active compounds within the chocolate mass depended on their physico-chemical properties and the mass composition, such as fat content and crystallization state of the sugar particles. The compound accumulation in the fat phase increased with decreasing compound polarity and increasing fat content. In the presence of cocoa particles, a 1.5-fold fat content resulted in a 1.6-fold higher proportional acetic acid concentration in the fat phase. Further, total acetic acid concentrations raised in all model systems containing crystalline sugar or cocoa particles (by 13.8-56.9 %), indicating the formation of free acetic acid., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

A pilot plant extraction of cocoa bean shell CBS oligo- (hCHO) and polysaccharide (CHO) extracts using alkali isolation (0.5 M KOH) and a commercially available multi-enzymatic product, Depol™ 670L, was performed. Comparisons between laboratory and pilot scale recovery yields and saccharide profiles were assessed. Enrichment of chocolate-based formulations, composed of cocoa and carob, using CBS CHO extracts was investigated for its use as a functional food ingredient without compromising flavor. Sensory panels were held to assess formulations before and after enrichment by rating the intensity and likeness using a 9-point hedonic scale, where attributes consist of chocolate flavor, sweetness, sourness, bitterness, and melting in mouth. Overall, enriched formulations accounted for higher overall acceptance and purchase intent consumer ratings. Factor and boxplot analyses were applied to gain insight and perspective regarding the correlation and interaction between the intensity and likeness attributes, perceived complexity of the sensory attributes, as well as the mean quantitative descriptive responses specified by consumers., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Cabernet Sauvignon (CS) and a combination of Cabernet Sauvignon with blueberry extract (CS + B), were spray dried (using maltodextrin DE10, 13.5% w/w as a carrier) to obtain two types of phenolic-rich powders. The addition of blueberry to CS increased phenolic compounds content by 16%. Eight chocolate formulations were obtained by modifying concentrations of cocoa solids, cocoa butter, and sugar. Six of the samples were added with 10% w/w of phenolic-rich powder, while two of them remained as powder-free controls. The anthocyanin and flavan-3-ol profiles of chocolates were determined by HPLC-DAD-MS and HPLC-MS, respectively. In addition, the sensory dynamic profile of samples was assessed by Temporal Dominance of Sensations with a consumer panel. Results showed that the addition of phenolic-rich powders produced a significant increase in the anthocyanin composition obtaining the highest anthocyanin content in the white chocolate added with CS + B powder. On the other hand, adding 10% of CS powder to dark chocolate (55% cocoa pellets) did not result in a significant increase in phenolic compounds. The addition of phenolic-rich powders to chocolates influenced visual color, texture, and taste, leading to new products with distinctive characteristics and increasing the possibility of using phenolic-rich powders as innovative and healthy ingredients., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study is a bibliometric analysis and literature review on the use of oleogels (OGs), hydrogels (HGs) and hybrid gels (HYGs) in chocolate, compounds and spreads with the aim of reducing the saturated fat in these products. The articles were selected by analyzing titles, keywords and abstracts in the Web of Science (WoS), Scopus and Google Scholar databases. Supplementary documents were obtained from government sources, including patent registrations. The theoretical and practical aspects were critically analyzed, highlighting the main points of agreement and disagreement between the authors. The results revealed a lack of regulations and official guidelines that widely allow the use of OGs, HGs and HYGs in chocolate confectionery products. The type and characteristics of raw materials affect the properties of products. Replacing cocoa butter (CB) with OGs, HGs or HYGs also affects texture, melting point and behavior, and nutritional aspects. These substitutions can result in products with better sensory acceptance and health benefits, such as reducing saturated fat and promoting cardiovascular health. However, it is important to find the ideal combination and proportions of components to obtain the desired properties in the final products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A screening of 24 Chuncho cacao postharvest processes was carried out and three treatments were identified with the best levels of functional non-volatile metabolites and the best profile of volatile metabolites. The relationship between the sensory profile of flavor and aroma, volatile and non-volatile metabolites was investigated by multiblock analysis. The most prominent volatile metabolites were benzaldehyde (Bz), 2,3,5-trimethylpyrazine (TriMP), 2,3,5,6-tetramethylpyrazine (TetraMP), myrcene (Myr), limonene (LM), ethylphenyl acetate (EpHAc) and 2-phenylethyl acetate (pHEthAc), and functional nonvolatile metabolites were total flavonoids (Tflav), total polyphenols (TppH), catechins (Cat), epicatechin (EpCat), caffeine (Caf), and theobromine (TeoB). Sensory analysis by MFA (multiple factorial analysis) allowed checking the substrates of the metabolites and highlighting correlated flavor and aroma descriptors. Based on multiblock analysis, four new descriptors were identified for Bz, Myr, and LM, such as nut, astringent, bitter, and crude. For TetraMP the fruity flavor was identified and for TriMP it was identified as Malt flavor. For Cat, EpCat, Caf, TeoB, Tflav, and TppH, the descriptors bitter, astringent, floral, and fruity were identified. This study will allow Chuncho cocoa farmers and producers to have a cocoa paste production scheme with a high value of functional metabolites and sensory profile, and value this Peruvian accession., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

This study aims to evaluate the effect of sucrose replacer mixtures (erythritol, mannitol, or tagatose in combination with inulin or polydextrose) on the crystal morphology, particle size distribution, rheology, melting properties, and fat polymorphism of dark compound chocolate. The result showed that the replacer mixture's hygroscopicity, particle size, and sugar crystal shape might significantly impact dark compound chocolate's rheological and textural properties but had no substantial impact on the melting properties and fat crystallization. Mannitol-containing samples exhibited the highest rheological value, likely related to their high moisture content, small particle size, and elongated crystal shape. Due to the similar specific surface area and comparable D90 value, the sample containing erythritol-polydextrose mixture resulted in a similar (P ≥ 0.05) Casson yield value (46.184 ± 2.45 Pa) compared to the sample containing sucrose (38.348 ± 1.68 Pa). It could be a potential sucrose replacer in the dark compound chocolate., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Monash University Malaysia. Published by Elsevier Ltd.. All rights reserved.)

In this study, we discussed covalent and non-covalent reactions between cocoa polyphenols and proteins (milk and cocoa) and the possible effects of these reactions on their bioaccessibility, considering environmental and processing conditions. Better insight into these interactions is crucial for understanding the biological effects of polyphenols, developing nutritional strategies, and improving food processing and storage. Protein-polyphenol reactions affect the properties of the final product and can lead to the formation of various precursors at various stages in the manufacturing process, such as fermentation, roasting, alkalization, and conching. Due to the complex composition of the chocolate and the various technological processes, comprehensive food profiling strategies should be applied to analyze protein-polyphenol covalent reactions covering a wide range of potential reaction products. This will help to identify potential effects on the bioaccessibility of bioactive compounds such as low-molecular-weight peptides and polyphenols. To achieve this, databases of potential reaction products and their binding sites can be generated, and the effects of various process conditions on related parameters can be investigated. This would then allow to a deeper insight into mechanisms behind protein-polyphenol interactions in chocolate, and develop strategies to optimize chocolate production for improved nutritional and sensory properties.

In recent years, gut microbiome has evolved as a focal point of interest with growing recognition that a well-balanced gut microbiota is highly relevant to an individual's health status. The present review provides a mechanistic insight on the effects of cocoa chemicals on the gut microbiome and further reveals in silico biomarkers, taxonomic and functional features that distinguish gut microbiome of cocoa consumers and controls by using 16S rRNA gene sequencing data. The polyphenols in cocoa can change the gut microbiota either by inhibiting the growth of pathogenic bacteria in the gut such as Clostridium perfringens or by increasing the growth of beneficial microbiota in the gut such as Lactobacillus and Bifidobacterium. This paper demonstrates the holistic effect of gut microbiota on cocoa chemicals and how it impacts human health. We present herein the first comprehensive review and analysis of how raw and roasted cocoa and its products can specifically influence gut homeostasis, and likewise, how microbiota metabolizes cocoa chemicals. In addition to that, our 16S rRNA amplicon sequencing analysis revealed that the flavone and flavonols metabolism, aminobenzoate degradation and fatty acid elongation pathways represent the three most important signatures of microbial functions associated with cocoa consumption.

Mass spectrometry has been widely accepted as a confirmatory tool for the sensitive detection of undeclared presence of allergenic ingredients. Multiple methods have been developed so far, achieving different levels of sensitivity and robustness, still lacking harmonization of the analytical validation and impairing comparability of results. In this investigation, a quantitative method has been validated in-house for the determination of six allergenic ingredients (cow's milk, hen's egg, peanut, soybean, hazelnut, and almond) in a chocolate-based matrix. The latter has been produced in a food pilot plant to provide a real and well-characterized matrix for proper assessment of method performance characteristics according to official guidelines. In particular, recent considerations issued by the European Committee for Standardization have been followed to guide a rigorous single-laboratory validation and to feature the main method performance, such as selectivity, linearity, and sensitivity. Synthetic surrogates of the peptide markers have been used both in native and labelled forms in matrix-matched calibration curves as external calibrants and internal standards, respectively. A two-order of magnitude range was investigated, focusing on the low concentration range for proper assessment of the detection and quantification limits (LOD and LOQ) by rigorous calibration approach. Conversion factors for all six allergenic ingredients have been determined for the first time to report the final quantitative information as fraction of total allergenic food protein (TAFP) per mass of food (µg TAFP /g food ), since such a reporting unit is exploitable in allergenic risk assessment plans. The method achieved good sensitivity with LOD values ranging between 0.08 and 0.2 µg TAFP /g food , for all ingredients besides egg and soybean, whose quantitative markers reported a slightly higher limit (1.1 and 1.2 µg TAFP /g food , respectively). Different samples of chocolate bar incurred at four defined concentration levels close to the currently available threshold doses have been analyzed to test the quantitative performance of the analytical method, with a proper estimate of the measurement uncertainty from different sources of variability. The sensitivity achieved resulted in compliance with the various threshold doses issued or recommended worldwide., (© 2023. The Author(s).)

Because food flavor is perceived through a combination of odor and taste, an analytical method that covers both dimensions would be very beneficial for mapping the consistent product quality over the entirety of a manufacturing process. Such a method, so-called "unified flavor quantitation", has been successfully applied to several different food products in recent years. The simultaneous detection of aroma and taste compounds by means of ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) enables the analysis and quantification of an enormously large number of compounds in a single run. To evaluate the limits of this method, chocolate, a high-fat, complex matrix, was selected. In 38 distinct commercial chocolate samples, 20 flavor-active acids, aldehydes, and sugars were analyzed after a simple, rapid extraction step followed by derivatization with 3-nitrophenylhydrazine using a single UHPLC-MS/MS method. The results obtained highlight the great potential of the "unified flavor quantitation" approach and demonstrate the possibility of high-throughput quantitation of key aroma- and taste-active molecules in a single assay.

The main component of chocolate, cocoa (Theobroma cacao), is a significant commercial agricultural plant that directly sustains the livelihoods of an estimated forty to fifty million people. The economies of many cocoa producing nations, particularly those in the developing world, are supported by cocoa export revenue. To ensure satisfactory yields, however, the plant is usually intensely treated with pesticides because it is vulnerable to disease and pest attacks. Even though pesticides help protect the cocoa plant, unintended environmental contamination is also likely. Honey, produced from nectar obtained by honeybees from flowers while foraging, can serve as a good indicator for the level of pesticide residues and environmental pesticide build-up in landscapes. Here, we use a systematic literature review to quantify the extent of research on residues of pesticides used in cocoa cultivation in honey. In 81% of the 104 studies examined for this analysis, 169 distinct compounds were detected. Imidacloprid was the most frequently detected pesticide, making neonicotinoids the most frequently found class of pesticides overall. However, in cocoa producing countries, organophosphates, organochlorines, and pyrethroids were the most frequently detected pesticides. Interestingly, only 19% of studies were carried out in cocoa producing countries. We recommend prioritizing more research in the countries that produce cocoa to help to understand the potential impact of pesticide residues linked with cocoa cultivation in honey and the environment more generally to inform better pesticide usage, human health, and environmental policies., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Boakye et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Aiming to improve the quality of cocoa, preconditioning of cocoa after harvesting and before fermentation has become an on-farm processing step of great interest in recent times. The present work aimed to evaluate the influence of a pre-drying process on the volatile composition of Ecuadorian bulk (Forastero and CCN-51) and fine-flavour (ETT103 and LR14) cocoa at the end of primary processing. A total of 63 volatile compounds including aldehydes, alcohols, acids, ketones, esters, terpenes, lactones and other miscellaneous compounds were identified in cocoa samples by HS-SPME-GC-MS. The use of a pre-drying step revealed a varietal homogenization and a reduction in the fermentation time, making this preconditioning step an inexpensive and attractive option for farmers. Moreover, different varietal behaviour was observed after pre-drying, the fine-flavour varieties obtaining a clear improvement in aromatic quality with higher levels of compounds imparting positive notes., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Roasting is an important unit operation for the development of characteristic chocolate aroma during manufacturing. However, there is an increase in interest in minimally processed chocolate products due to their potential positive health benefits. The odor-important compounds and sensory characteristics of minimally processed (unroasted) and conventionally roasted dark chocolates were determined by gas chromatography-olfactometry, aroma extract dilution analysis (AEDA), and stable isotope dilution analysis (SIDA). Except for acetic acid, all odorants had higher odor-activity values (OAVs) in roasted chocolate. Acetic acid, developed during fermentation and drying, had the highest OAV in both chocolates but was better preserved in unroasted chocolate. Compounds making a greater aroma impact on roasted chocolate compared with unroasted chocolate included dimethyl trisulfide, 2-ethyl-3,5-dimethylpyrazine, and 3-methylbutanal. Nine significant sensory attributes in unroasted and roasted chocolates were identified. Vinegar (aroma) and roasted (aroma and aroma by mouth), sweet (taste), and hardness (texture) attributes differed between unroasted and roasted chocolates. The results of this study enforce the embracement of low thermal processes to showcase the inherent flavor potential of cacao beans but also to support the concept of chocolate "terroir" by potentially preserving important aroma compounds developed during fermentation.

Rapid assessment of pesticide residues ensures cocoa bean quality and marketability. In this study, a portable FTIR instrument equipped with a triple reflection attenuated total reflectance (ATR) accessory was used to screen cocoa beans for pesticide residues. Cocoa beans (n = 75) were obtained from major cocoa growing regions of Peru and were quantified for pesticides by gas chromatography (GC) or liquid chromatography (LC) coupled with mass spectrometry (MS). The FTIR spectra were used to detect the presence of pesticides in cocoa beans or lipid fraction (butter) by using a pattern recognition (Soft Independent Modeling by Class Analogy, SIMCA) algorithm, which produced a significant discrimination for cocoa nibs (free or with pesticides). The variables related to the class grouping were assigned to the aliphatic (3200-2800 cm -1 ) region with an interclass distance (ICD) of 3.3. Subsequently, the concentration of pesticides in cocoa beans was predicted using a partial least squares regression analysis (PLSR), using an internal validation of the PLRS model, the cross-validation correlation coefficient (R val  = 0.954) and the cross-validation standard error (SECV = 14.9 mg/kg) were obtained. Additionally, an external validation was performed, obtaining the prediction correlation coefficient (R pre  = 0.940) and the standard error of prediction (SEP = 16.0 μg/kg) with high statistical performances, which demonstrates the excellent predictability of the PLSR model in a similar real application. The developed FTIR method presented limits of detection and quantification (LOD = 9.8 μg/kg; LOQ = 23.1 μg/kg) with four optimum factors (PC). Mid-infrared spectroscopy (MIR) offered a viable alternative for field screening of cocoa., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Luis E. Rodriguez-Saona reports financial support was provided by Centro de Innovacion del Cacao., (Copyright © 2023 Elsevier B.V. All rights reserved.)

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

The physical characterization of chocolate products is vital in manufacturing, and the chocolate's processing time and composition directly influence physical properties, such as rheology and melting. The objective of this study was to investigate the effects of processing time and the ingredient types on the physical properties of milk chocolates containing soy milk powder and soy protein isolate. Characteristics of skimmed milk chocolate (SMC) and whole milk chocolate (WMC) were compared to soy milk chocolate and soy protein chocolate (SPC). Rheological data of chocolate products were fitted to Casson, Herschel-Bulkley, and Bingham models. The highest viscosity was observed for SPC samples, whereas yield stress was the highest for SMC samples after 2 hr of processing. The increase in milk and soy fats in the formulation softened the texture and decreased the whiteness index significantly (p < .05). PSD results show that SPC had the highest D90 (40.1 μm) and the lowest specific surface area (893 m 2 /kg) after 6 hr of processing. SPC samples had the narrowest particle size distribution observed by the span values. X-ray diffraction analysis showed that all the samples had the desired Form V, crystal form. The differential scanning calorimetry thermogram was used to determine phase transitions and melting behaviors. At the end of processing, melting enthalpies (ΔH melt ) were significantly lower (p < .05) in milk chocolates., (© 2023 Wiley Periodicals LLC.)

For the first time, the identification and quantification of trace level of new psychoactive substances (NPS) in a complex chocolate matrix have been reported. Since the beginning of 2022, suspected NPS-infused chocolate samples confiscated in inbound packages have been continuously sent to our laboratory for analysis. The qualitative gas chromatography-mass spectrometry (GC-MS) results were verified by 1 H nuclear magnetic resonance ( 1 H NMR) and 19 F NMR to distinguish between potential aromatic isomers. A total of 11 NPS including deoxymethoxetamine, 3-OH-PCP, 6-APB, 4-APB, 4-OH-MiPT, 3-FEA, 2-FEA, 3-MMC, bromazolam, 2-FDCK, and ADB-BUTINACA were detected in 65 seized chocolate samples. A general 1 H quantitative NMR ( 1 H qNMR) method for quantification of 297 types of NPS in complex chocolate matrixes was devised for the first time after rigorous analysis of various critical features of merit, including suitable deuterated solvent, internal standard, quantitative peaks, and instrument acquisition parameters. Validation of the method using six different types of NPS afforded limits of detection of 0.05-0.1 mg/mL, limits of quantification of 0.01-0.03 mg/mL, repeatability and reproducibility lower than 0.5% and 3.6%, recoveries of 91.7%∼104.4%, and absence of matrix effect. The quantitative analysis of 65 seized chocolate samples by 1 H qNMR and 19 F qNMR showed that the content of NPS was in the range of 0.5 mg/g∼44.1 mg/g. Generally, the developed qNMR method was simple, fast, precise, and can be performed without reference materials of NPS. Since the type and content of NPS are relatively random, chocolate consumers will face huge health risks. Therefore, this new trend of NPS-infused chocolate deserves and requires more attention from national NPS monitoring departments as well as forensic laboratories., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Cocoa powder is a highly consumed product all over the world which could be substituted by cheaper raw materials resulting in food fraud. In this work, a non-targeted metabolomics approach based on the use of reversed-phase liquid chromatography coupled to high-resolution mass spectrometry was developed to carry out the characterization of cocoa powder samples adulterated, at two different levels, with carob flour, soy flour, and chicory. The sample preparation protocol and the chromatographic parameters were optimized to extract and detect the highest number of molecular features. Both non-supervised and supervised statistical methods were employed to analyze the most significant variables that gave rise to group discrimination. From the 21 and 37 significant variables analyzed in positive and negative ionization modes, respectively, a total of 20 were tentatively identified. Different families of compounds including flavonoids, fatty acids, terpenoids, lysophospholipids, and a galactolipid could be pointed out as cocoa adulteration markers., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

A non-segregated kinetic model is proposed to describe a fermentation process of agro-industrial residues derived via cocoa (mucilage juice) by Pichia kudriavzevii . The novel proposed hybrid model is based on a multiple coupling reaction mechanisms (structured) to describe the kinetics of substrate consumption, biomass, carbon dioxide, and ethanol, coupled to an unstructured model for the activity enzyme. The parameters of the kinetic model are estimated by non-linear least-squares curve fitting using the Marquardt-Levenberg algorithm. In addition, numerical simulations were compared with the experimental data via residual graphs. The effectiveness of the model was statistically evaluated using dimensionless efficiency coefficients under different initial conditions. A global sensitivity analysis was applied (Fisher's information matrix). The experimental results of the batch reactor showed a maximum ethanol concentration of 29 g/L, with a yield of 0.48 g-ethanol/g-glucose and a productivity of 0.30 g/L h. The method determined that the cell formation coefficient and the specific substrate consumption rate ( θ 1 and θ 2 ) directly influence most of the states of our system. The proposed scheme is particularly suitable to assist in the rational design of cell factory properties or fermentation processes because it can represent the complex biochemistry in more detail and under different initial experimental conditions; the above reveals that the generated model is robust and can be considered for control and optimization purposes.

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

The high incidence of aflatoxins (AFs) in chocolates suggests the necessity to create a practical and cost-effective processing strategy for eliminating mycotoxins. The present study aimed to assess the adsorption abilities of activated charcoal (A. charcoal), yeast ( Saccharomyces cerevisiae ), and the probiotic Lactobacillus rhamnosus as AFs adsorbents in three forms-sole, di- and tri-mix-in phosphate-buffered saline (PBS) through an in vitro approach, simulated to mimic the conditions present in the gastrointestinal tract (GIT) based on pH, time and AFs concentration. In addition, the novel fortification of chocolate with A. charcoal, probiotic, and yeast (tri-mix adsorbents) was evaluated for its effects on the sensory properties. Using HPLC, 60 samples of dark, milk, bitter, couverture, powder, and wafer chocolates were examined for the presence of AFs. Results showed that all the examined samples contained AFs, with maximum concentrations of 2.32, 1.81, and 1.66 µg/kg for powder, milk, and dark chocolates, respectively. The combined treatment demonstrated the highest adsorption efficiency (96.8%) among all tested compounds. Scanning electron microscope (SEM) analysis revealed the tested adsorbents to be effective AF-binding agents. Moreover, the novel combination of tri-mix fortified chocolate had a minor cytotoxicity impact on the adsorptive abilities, with the highest binding at pH 6.8 for 4 h, in addition to inducing an insignificant effect on the sensory attributes of dark chocolate. Tri-mix is thus recommended in the manufacturing of dark chocolate in order to enhance the safety of the newly developed product.

Green analysis is defined as the analysis of chemicals in a manner where sample extraction and analysis are performed with least amounts of steps, low hazardous materials, while maintaining efficiency in terms of analytes detection. Coffee and cocoa represent two of the most popular and valued beverages worldwide in addition to their several products i.e., cocoa butter, chocolates. This study presents a comprehensive overview of green methods used to evaluate cocoa and coffee seeds quality compared to other conventional techniques highlighting advantages and or limitations of each. Green techniques discussed in this review include solid phase microextraction, spectroscopic techniques i.e., infra-red (IR) spectroscopy and nuclear magnetic resonance (NMR) besides, e-tongue and e-nose for detection of flavor. The employment of multivariate data analysis in data interpretation is also highlighted in the context of identifying key components pertinent to specific variety, processing method, and or geographical origin., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Fine flavor properties of chocolates such as fruity, floral, and cocoa-like were decoded on a molecular level for the first time. The molecular compositions of six chocolates made out of liquors that were referenced with specific sensory attributes were analyzed. After the screening for odor-active molecules by aroma extract dilution analysis, selected compounds were quantitated with the overall aim to decode the distinct fine flavor attributes on a molecular level. Acidic and fruity flavor notes were associated with high dose over threshold factors (DoT factors) of acetic acid and fruity smelling esters such as ethyl 2-methylbutanaote, ethyl 3-methylbutanoate, and 3-methylbutyl acetate, respectively. Cocoa-like and roasty flavor notes were associated with high DoT factors for 2-methylbutanal, 3-methylbutanal, 4-hydroxy-2,5-dimethylfuran-3(2 H )-one, and dimethyltrisulfane. The floral and astringent flavors were linked to high DoT factors of (-)-epicatechin, procyanidin B2, procyanidin C1, and 2-phenylethan-1-ol.

A total of 56 key volatile compounds present in natural and alkalized cocoa powders have been rapidly evaluated using a non-target approach using stir bar sorptive extraction gas chromatography mass spectrometry (SBSE-GC-MS) coupled to Parallel Factor Analysis 2 (PARAFAC2) automated in PARADISe. Principal component analysis (PCA) explained 80% of the variability of the concentration, in four PCs, which revealed specific groups of volatile characteristics. Partial least squares discriminant analysis (PLS-DA) helped to identify volatile compounds that were correlated to the different degrees of alkalization. Dynamics between compounds such as the acetophenone increasing and toluene and furfural decreasing in medium and strongly alkalized cocoas allowed its differentiation from natural cocoa samples. Thus, the proposed comprehensive analysis is a useful tool for understanding volatiles, e.g., for the quality control of cocoa powders with significant time and costs savings., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In this work, a rapid detection method using solid sampling electrothermal vaporization atomic absorption spectrometry (SS-ETV-AAS) was established for cadmium in chocolate. The instrumental system includes a solid sampling ETV unit, a catalytic pyrolysis furnace, an AAS detector, and a gas supply system with only an air pump and a hydrogen generator. Herein, MgO material with 1.0−1.5 mm particle size was first employed to replace the kaolin filler previously used to further shorten the peak width and to thereby improve the sensitivity. With 350 mL/min of air, a chocolate sample was heated for 25 s from 435 to 464 °C to remove water and organic matrices; then, after supplying 240 mL/min hydrogen and turning down air to 120 mL/min, a N2/H2 mixture gas was formed to accelerate Cd vaporization from chocolate residue under 465 to 765 °C. Under the optimized conditions, the detection limit (LOD) was obviously lowered to 70 pg/g (vs. previous 150 pg/g) with R2 > 0.999; the relative standard deviations (RSD) of repeated measurements for real chocolate samples ranged from 1.5% to 6.4%, indicating a favorable precision; and the Cd recoveries were in the range of 93−107%, proving a satisfied accuracy. Thus, the total analysis time is less than 3 min without the sample digestion process. Thereafter, 78 chocolate samples with different brands from 9 producing countries in China market were collected and measured by this proposed method. Based on the measured Cd concentrations, a dietary exposure assessment was performed for Chinese residents, and the target hazard quotient (THQ) values are all less than 1, proving no significant health risk from intaking chocolate cadmium for Chinese residents.

Background: Chocolate is a rich source of essential and non-essential elements. A new liquid-liquid extraction (LLE) approach, extraction induced by emulsion breaking (EIEB), is proposed in which the analyzed elements are transferred from the organic phase to the aqueous phase before measurement by flame atomic absorption spectrometry (FAAS)., Objective: To compare EIEB to microwave digestion (MWD) for extraction of elements from chocolate prior to FAAS., Methods: EIEB parameters were varied to optimize the procedure. EIEB-FAAS was then compared to MWD-FAAS for the analysis of Ca, Cu, Fe, Mn, Ni, and Zn in milk and dark chocolate samples. A certified reference material (NIST 2384, baking chocolate) was analyzed to determine the recoveries of Ca, Cu, Fe, Mn, and Zn by the two methods., Results: The optimized EIEB extraction method involves dilution of tempered chocolate with toluene, ultrasonic emulsification with acidified Triton X-114, breaking the emulsion by heating, and centrifugation to produce two well-defined phases. Analysis of dark and milk chocolate samples showed similar repeatability by EIEB-FAAS (RSDr 0.3 to 6.6% in dark and 0.5 to 8.7% in milk) and MWD-FAAS (RSDr 0.5 to 5.4% in dark and 0.7 to 10.2% in milk), with no significant difference detected between the methods for analysis of Ca, Cu, Fe, Mn, Ni, and Zn based on Student's t-test. Analysis of NIST 2384 baking chocolate certified reference material for Ca, Cu, Fe, Mn, and Zn demonstrated recoveries of 98.6 to 99.5% for EIEB-FAAS compared to 95.8 to 98.6% for MWD-FAAS., Conclusion: EIEB-FAAS was shown to provide high recovery and excellent repeatability for accurate determination of Ca, Cu, Fe, Mn, Ni, and Zn from dark and milk chocolates., Highlight: The EIEB-FAAS method is simpler and requires fewer reagents compared to other sample preparation methods and allows the calibration to be carried out using aqueous calibration solutions., (© The Author(s) 2022. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

A methodology for simultaneous (not sequential) determination of Cd and Ni in powder chocolate samples by high-resolution continuum source graphite furnace atomic absorption spectroscopy (HR-CS-GF-AAS) and slurry sample analysis was developed. Acid digestion of the samples was not necessary, considering the high total fat content of this type of matrix (above 10 %), and no apparent deterioration of the graphite tubes was even observed once these samples were subjected to analysis. Despite the volatilities of both elements are quite different, simultaneous quantification was achieved using a slow heating ramp atomization, covering a convenient and wide temperature gradient for Cd volatilization at low temperatures and subsequent atomization of Ni at elevated temperatures, using the same heating program without an apparent change in sensitivity. This slow heating ramp was essayed between 200 and 1500 °C·s -1 and optimized at 400 °C·s -1 , achieving the simultaneous measuring of Cd in the principal line (228.8018 nm) and using a secondary for Ni (228.9984 nm). No stop events for sequential changes in wavelength or temperature additional steps were involved in the method and molecular spectral interferences overlapping the analyte signals were corrected appropriately by a pragmatic strategy taking advantage of the fine structure signals adjacent to analyte signals within the same spectrum recorded. Palladium matrix modifier and aqueous standards were used for the calibration and determination of both elements in the samples. The method accuracy was successfully confirmed using a reference material (NIST SRM 1573a). An improvement in the sensitivity of Ni was found using a greater number of pixels for signal integration, central and five adjacent pixels (CP ± 5), while a lengthening of the linear range of Cd was obtained by using an attenuated signal (±2) or using only one pixel (CP). The limits of detection were 0.027 μg g -1 and 0.22 μg g -1 for Cd and Ni respectively by this simple methodology using aqueous standards for calibration and the simultaneous determination of both elements in the same measurement. The respective limits of quantification for Cd and Ni were 0.090 and 0.72 μg g -1 and the relative standard deviations in samples (n = 5) were about 4-20% and 0.3-11% for Cd and Ni respectively. The optimized method was used for the determination of both elements in real samples purchased from retail supermarkets finding variable concentrations between 0.5 and 5.7 μg g -1 for Cd and 2.1-10.9 μg g -1 for Ni, comparable to levels reported in the literature referred to elsewhere in the world., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the volatile compounds of chocolates made of Brazilian cocoas and statistically track them according to the products' sensorial profile in order to relate them to consumers' acceptance by preference map methodology. The intensity of the chocolate, acidity, woody, smoked, green, floral, burned, musty, and cocoa notes from chocolates produced with cocoa from different Brazilian states were analyzed by a trained panel and by 128 consumers. Samples from Côte d'Ivoire, which is known for its high-quality chocolate, were evaluated for comparison. Solid-phase microextraction headspace sampling/gas chromatography-mass spectrometry was employed to evaluate the samples' volatile compounds. One hundred volatile compounds were identified within the samples. The results from the preference maps showed that the maximum preference was found for chocolate made of cocoa from Rondônia, Bahia, and Espírito Santo and Côte d'Ivoire and organic samples from Pará. The ideal sample point was characterized by intense chocolate, floral, and woody notes and mild green and burned notes. The presence of furfural, 3-methyl butanal, phenethyl acetate, 2-phenyl-5-methyl-2-hexenal, methyl pyrazine, phenethyl acetate, 2-phenyl-5-methyl-2-hexenal, and tetramethyl pyrazine were shown to be important for consumer acceptance in the ideal product, whereas the presence of (Z)-2-heptenal and 2-pentyl furan may increase consumer rejection. 2,3-Methyl pyrazine, methyl pyrazine, and 2,3-butanediol, which are important volatile compounds previously reported in the literature, were statistically tracked to both positive and negative sample attributes and must be better explored concerning consumers' acceptance of chocolates., (Copyright © 2022 Elsevier Ltd. All rights reserved.)