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Since 2019, EU limits apply to cadmium (Cd) concentrations in cacao-derived food products. The dietary risk assessment leading to that regulation used consumption surveys aggregated to a limited number of chocolate product categories and did not consider differences in Cd bioaccessibility. Here, the cacao-related dietary Cd exposure in the Belgian population was estimated with higher resolution and accounting for bioaccessibility. A food frequency questionnaire and a 24-h recall (N = 2055) were set up for the Belgian population, in combination with ICP-MS analysis of a large subset of cacao-containing products (N = 349). Both the average chocolate consumption (28 g day−1) and the relative contribution of chocolate to the total dietary Cd exposure (7–9%) were higher than previously estimated for the Belgian population, probably because of some selection bias towards chocolate consumers in the cohort. The Cd bioaccessibility in chocolate products was a factor 5 (cacao powder) and 2 (dark chocolate) lower compared to wheat flour, suggesting lower bioavailability in chocolate than in wheat, which is a main contributor to dietary Cd. This study suggests that Cd intake from cacao consumption has been underestimated because of hidden cacao in non-chocolate food categories but, in contrast, may have overestimated the true exposure because of lower bioavailability compared to the main foodstuffs contributing to Cd exposure. • Chocolate-related Cd intake is underestimated in dietary Cd exposure surveys. • Cd bioaccessibility in chocolates decreases with increasing cacao content. • Cd bioaccessibility in cacao products is up to a factor 5 lower compared to wheat. [ABSTRACT FROM AUTHOR]

• Cacao bean incubation reveals that fermentation can reduce nib Cd content. • Reduction of nib Cd was most pronounced at high temperature and low pH. • Nib Cd mobilisation during fermentation is likely related to phytate breakdown. Earlier studies revealed that cadmium (Cd) concentrations in cacao nibs can decrease by a factor up to 1.3 during fermentation. Here, fermentation was mimicked by incubating beans at different temperatures, and acetic acid and ethanol concentrations in the incubation media. Nib Cd concentrations decreased during incubation by mobilisation in the nibs and subsequent outward migration to the testa and the incubation solution. This was most pronounced when high concentrations of acetic acid were combined with high temperature, while ethanol had no statistically significant effect. Incubation under typical fermentation conditions (45 °C and 20.0 g acetic acid L−1) reduced the nib Cd concentration by a factor 1.3. This factor increased to 1.6 under more extreme conditions, i.e. 65 °C and 40 g acetic acid L−1. The final nib Cd concentrations correlated well to nib phytate concentrations (R2 = 0.56), suggesting hydrolysis of phytate and mobilisation of the associated Cd2+. [ABSTRACT FROM AUTHOR]

Previous work has shown that cacao nib cadmium (Cd) concentrations decrease during fermentation, but only when reaching sufficiently low nib pH. In this work, lab-scale experiments (5 kg units) with lactic and acetic acid amendments were ineffective at reducing the total nib Cd concentration. In contrast, the water-extractable fraction of the nib Cd concentration clearly increased when the pH was decreased. When single pod derived beans were embedded inside a full-scale fermentation box to monitor the effect of the fermentation effect with high precision, nib Cd concentrations decreased by a factor 1.25 (P-value <0.05) after four days of fermentation. Visualisation of the mobile Cd gradient within beans with LA-ICP-MS (using imprints of transversal cuts exposed to a metal binding gel) revealed that fermentation enhances the Cd mobility in the nibs. • Fermentation induces outward migration of Cd from cacao nib to mucilage and testa. • Water extractable Cd in cacao nibs increases with decreasing pH. • Organic acid treatments during lab scale fermentation did not lower nib Cd content. • Mobile Cd gradients within cacao beans were visualized with LA-ICP-MS. [ABSTRACT FROM AUTHOR]

• Cadmium concentrations are highest in cacao testa followed by nibs and mucilage. • LA-ICP-MS was used successfully to visualize the distribution of Cd in cacao. • X-ray absorption spectroscopy showed Cd bound to O/N-ligands in cacao nib and testa. • Extensive fermentation can reduce the Cd concentration in the final product. • Nib pH controls Cd migration to outer tissues during cacao fermentation. A large fraction of the South-American cacao production is affected by new cadmium (Cd) regulations in cacao. This work was set up to characterize the distribution and speciation of Cd within the cacao fruit and to monitor potential Cd redistribution during cacao fermentation. In cacao fruits from four locations, Cd concentrations decreased with testa > nib ~ placenta ~ pod husk > mucilage. The distribution of Cd within cacao beans was successfully visualized using laser ablation inductively coupled plasma spectrometry (LA-ICP-MS) and confirmed higher Cd concentrations in the testa than in the nib. Speciation analysis by X-ray absorption spectroscopy (XANES) of unfermented cacao beans revealed that Cd was bound to O/N-ligands in both nib and testa. Fermentation induced an outward Cd migration from the nibs to the testa, i.e. against the total concentration gradient. This migration occurred only if the fermentation was sufficiently extensive to decrease the pH in the nib to <5.0, likely as a result of increased Cd mobility due to organic acid penetration into the nibs. The change in dry weight based nib Cd concentrations during fermentation was, on average, a factor 1.3 decrease. We propose that nib Cd can be reduced if the nib pH is sufficiently acidified during fermentation. However, a balance must be found between flavor development and Cd removal since extreme acidity is detrimental for cacao flavor. [ABSTRACT FROM AUTHOR]

• Multi element ICP-MS analysis of 139 single origin chocolates. • Cacao content and cacao origin determine trace element levels of chocolates. • Decision tree analysis (CART) separates chocolates according to cacao origin. Commercially available single origin chocolates (n = 139) were analysed by ICP-MS to identify the potential of elemental fingerprinting for tracing cacao origin in chocolate and to compare chocolate composition relative to trace metal limits. Cadmium (Cd) concentrations exceeded the EU limit of 0.80 mg kg−1 in 16 samples, all produced with cacao from South or Central America. Six samples contained lead (Pb) concentrations >0.10 mg kg−1, the limit of the Codex Alimentarius for edible fats. Increasing cacao content was associated with increased element concentrations for most elements, indicating cacao as the main source of minerals and trace elements. Significant differences in elemental composition between origins (P value ≤ 0.05) were found for Ba, Cd, Mo and Sr. Classification and regression tree analysis (CART) resulted in a decision tree that could effectively classify chocolate samples by cacao origin continent (overall misclassification rate 23%) based on the concentrations of five elements (Ba, Cd, Mo, Sr and Zn). Samples of South America were classified based on their Cd concentration, indicating the geogenic origin of Cd. [ABSTRACT FROM AUTHOR]

• Cadmium is complexed by sulphur ligands in cacao nibs at high Cd concentrations. • At lower nib Cd concentrations, Cd is complexed by oxygen ligands. • Cadmium in cacao nibs becomes complexed by oxygen ligands after incubation. • Changes in Cd speciation are driven both by acidification and phytate breakdown. Cadmium (Cd) concentrations in cacao often exceed food limits. Recently, it was shown that cacao bean fermentation enhances Cd solubility, opening potential for Cd mitigation in cacao products. This study was set-up to identify changes in Cd speciation during fermentation. X-Ray absorption spectroscopy (XAS) complemented with speciation calculations, were used on samples collected from high and low Cd farms, that were subjected to a fermentation-like incubation that reached high temperatures (>45 °C) and acidic pH (<5). Incubation decreased nib Cd concentration up to a factor 1.5 and changed Cd complexation in high Cd beans from sulphur to oxygen ligands, likely due to pH changes. In beans with lower Cd concentrations, Cd was complexed before and after incubation with oxygen-ligands. A combination of pH changes and/or phytate breakdown may explain the migration of Cd outward from the nib. XAS and speciation calculations proved complimentary techniques and indicated similar speciation changes during fermentation. [ABSTRACT FROM AUTHOR]

Abstract Recent cadmium (Cd) regulation in chocolate threatens the sustainability of cacao production in Southwest America. Cadmium contamination in cacao beans has not been assessed at a country level. A nationwide survey was conducted in Ecuador to identify the spatial distribution of Cd in cacao beans, as well as soil and agronomic factors involved. Paired soil and plant samples (pods and leaves) were collected at 560 locations. Information on agronomic practices was obtained through a prepared questionnaire for farmers. Total soil Cd averaged 0.44 mg kg−1 which is typical for young and non-polluted soils. Mean Cd concentration in peeled beans was 0.90 mg kg−1 and 45% of samples exceeded the 0.60 mg kg−1 threshold. Bean Cd hotspots were identified in some areas in seven provinces. Multivariate regression analysis showed that bean Cd concentrations increased with increasing total soil Cd and with decreasing soil pH, oxalate-extractable manganese (Mn ox) and organic carbon (OC) (R2 = 0.65), suggesting that Cd solubility in soil mainly affects Cd uptake. Bean Cd concentration decreased a factor of 1.4 as the age of the orchard increased from 4 to 40 years. Bean Cd concentration was inconsistently affected by genotype (CCN-51 vs. Nacional), pruning or application of fertilizers. It is concluded that the relatively larger bean Cd concentrations in Ecuador are related to the high Cd uptake capacity of the plants combined with their cultivation on young soils, instead of Cd depleted weathered soils. Mitigation strategies should consider the application of amendments to modify such soil properties to lower soil Cd availability. There is scope for genetic mitigation strategy to reduce bean Cd, but this needs to be properly investigated. Graphical abstract Unlabelled Image Highlights • Spatial distribution of Cd concentrations in cacao beans is proposed for Ecuador. • 45% of samples exceeded the 0.60 mg kg−1 (dry weight bases) threshold. • Total soil Cd, pH and % organic carbon are the main factors affecting Cd concentration in beans. • Soil to bean transfer factors suggest a high affinity of cacao plants for Cd uptake. • Cd hotspots were identified in several cocoa producing regions of Ecuador. [ABSTRACT FROM AUTHOR]

Background and aim: Understanding the maturation of upper limb (UL) movement characteristics in typically developing (TD) children is key to explore UL deficits in those with neurodevelopmental disorders. Three-dimensional motion analysis (3DMA) offers a reliable tool to comprehensively evaluate UL motion. However, studies thus far mainly focused on specific pre-defined parameters extracted from kinematic waveforms. Here, we investigated age-related differences in UL movement characteristics over the entire movement cycle in TD children. Participants and methods: We assessed the non-dominant UL of 60 TD children (mean age 10y3m±3y1m) using 3DMA during eight tasks: reaching (forwards (RF), upwards (RU), sideways (RS)), reach-to-grasp (sphere (RGS), vertical cylinder (RGV)) and activities-of-daily-living mimicking tasks (hand-to-head (HTH), hand-to-mouth (HTM), hand-to-shoulder (HTS)). We investigated differences between four age-groups (5-7y, 8-10y, 11-12y, 13-15y) in: (1) spatiotemporal parameters (movement duration, peak velocity, time-to-peak velocity and trajectory straightness), and (2) 12 UL joint angles, using Statistical Parametric Mapping (SPM). Results: We found that the 5-7y children moved with lower peak velocity and less straight trajectories compared to the 11-12y group (peak velocity: RS, HTS, p<0.01; trajectory: RU, RS, RGV, HTS, p<0.01) and the 13-15y group (peak velocity: RF, RS, RGS, RGV, HTH, HTS, p<0.01; trajectory, all tasks, p<0.01). The 5-7y children showed increased scapular protraction compared to older children (8-10y and 11-12y, HTS), as well as increased scapular medial rotation compared to the 13-15y group (RGS). During RU, the 5-7y children moved more towards the frontal plane (shoulder), unlike the 13-15y group. Lastly, the 5-7y group used less elbow flexion than older children (11-12y and 13-15y) during HTH and HTS. Discussion and conclusion: In conclusion, our results point toward a maturation in UL movement characteristics up to age 11-12y, when UL motion seemed to reach a plateau. The reference values provided in this study will help to further optimize the interpretation of UL deficits in children with neurodevelopmental disorders. [ABSTRACT FROM AUTHOR]