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Background In February 2022, the United Kingdom (UK) detected a cluster of monophasic Salmonella Typhimurium based on whole genome sequencing (WGS). Subsequently, several countries reported cases belonging to this cluster. Epidemiological, microbiological and traceability investigations pointed toward a chocolate food business operator (FBO) in Belgium. We describe the magnitude of the outbreak, investigations performed and control measures taken. Methods Cases were ascertained based on internationally agreed case definitions and interviewed about food consumption prior to disease onset. Analytical epidemiological studies were conducted by the UK and Ireland. The Belgian food safety authority (FSA) coordinated microbiological and traceability investigations. Results A total of 456 cases (61% female), belonging to two genetically different WGS clusters, in 14 countries of the European Union, the UK, Switzerland, Canada and the United States were linked to the outbreak, between December 2021 and June 2022. 87% of cases were younger than 10 years. Brand A chocolate eggs, marketed for children, were reported as consumed by 168 cases (80%) with information. Analytical studies in the UK and Ireland showed a significantly higher odds of disease associated with consumption of brand A chocolate products. Retrospective investigations by the FSA revealed that routine samples (raw materials, intermediate, semi-finished and finished products as well as environmental and rinse oil samples) taken by the FBO linked to the Brand A products between December 2021 and January 2022 had tested positive for salmonella. Nine isolates were submitted to EnteroBase and matched with human isolates from both WGS clusters. The authorization for production was temporarily withdrawn on 8 April 2022 and all products of brand A were recalled worldwide, followed by a decrease in cases. Conclusions A multi-country salmonellosis outbreak linked to chocolate occurred in the months before Easter 2022. International collaboration between stakeholders from epidemiological, microbiological and food safety entities with rapid sharing of WGS results from human and nonhuman isolates were key in containing the outbreak. Implementation of routine WGS on human and nonhuman isolates will strengthen public health responses to future outbreaks. The magnitude of the outbreak underlines the importance of timely and open communication of FBOs to FSAs in case of salmonella detection.

Traditional booths where sensory evaluation usually takes place are highly controlled and therefore have limited ecological validity. Since virtual reality (VR) is substantially interactive and engaging, it has the potential to be applied in sensory science. In this preliminary study, three chocolate types (milk, white, and dark) were evaluated under three contextual settings, including sensory booths (control) and two VR environments (360-degree videos using VR headsets: (i) a pleasant sightseeing tour, and (ii) a live music concert). Untrained participants (n = 67) were asked to rate their liking and the intensity of different chocolate attributes based on the 9-point hedonic scale and just-about-right-scale (JAR). Emotions were evaluated using the check-all-that-apply (CATA) method. Results showed that there were no significant effects of context type on the tasting experience; however, there were significant effects of chocolate type. Milk and white chocolates were preferred over dark chocolate irrespective of the context type. Additionally, more positive emotions were elicited for the dark chocolate in the “virtual live concert” environment. Dark chocolate under the other two environments was associated with negative emotional terms, such as “bored” and “worried.” In terms of more reliable and ecologically valid sensory responses, further research is needed to match suitable VR environments to different chocolate types.

Cupuaçu [Theobroma grandiflorum (Wild ex Spreng.) K. Schum] seeds have been employed for a long time in the Amazon region for food purposes. Similar to cocoa, processed cupuaçu pulp and seeds can be used to produce juices, ice creams, confectionary products and cupulate®, which is a similar product to chocolate. However, its market penetration requires the mastery of all processing stages to improve the food quality and safety and to make possible an efficient technology transfer to the local small farmers and communities. Based on the above, the current research work aimed at monitoring and optimizing the consecutive fermentation and drying processes of cupuaçu seeds over 7 days each, as well as storage for 90 days. A greenhouse structure incorporating the fermenter and solar drying terrace was designed to be inexpensive, versatile, easily scalable, and easy to maintain and operate by the local small farmers after a short period of training. This research effort also aimed at giving a vision for the future creation of an integrative and sustainable cupuaçu system covering the economic, social, cultural and environmental vectors. The experimental design comprised 5 batches of 100 kg of seeds each. Several microbiological and physicochemical parameters were performed and correlated with processing variables. Microbiological parameters encompassed viable counts of mesophilic microorganisms, coliforms, yeasts, and molds, whereas physicochemical measures included fermentation and drying temperature, pH, acidity, dry matter, ashes, water activity, color, total proteins, lipids and carbohydrates, and energy. The average seed fermentation temperature varied from ca. 28 to 44 °C, reaching the maximum on day 3 and a final value of ca. 31 °C. Regarding solar drying, the average seed temperatures ranged from ca. 24 °C (at the end) to 39 °C on day 3, and an initial value of ca. 29 °C. The average final seed pH value of drying was 5.34 and was kept during storage. During storage, results demonstrated the existence of significant correlations among several experimental parameters under scrutiny. Finally, bean viable counts obtained during storage unfolded acceptable values of total mesophilic bacteria well below the maximum limit. Viable counts of yeast and molds were generally found between 3 and 4 log(CFU/gsample), and total coliforms were also detected, although both were at acceptable levels and well beneath the established maximum limits for food safety.

An ultrasound-assisted method for extraction of theobromine (TB) and caffeine (CF) from cacao seeds and chocolate products was developed. This extraction was carried out in 100 mL of heated water at 80 °C by using a 12.7-mm i.d. ultrasound probe and applying 240 W of power for 180 s. In order to quantify TB and CF, the aqueous extracts were treated with Carrez reagent to remove interferences. Then, 10 mL of the aqueous extract was adjusted at pH 12.5 and 10 mL of chloroform solvent was added. Ultrasound radiation was applied at 160 W of power for 30 s in order to transfer CF to the organic phase. The resulting emulsion was centrifuged at 6,000 rpm for 5 min to separate the phases. The photometric monitoring was at 276 nm for CF and at 273 nm for TB. This extraction method is simpler than the conventional liquid–liquid extraction which uses separatory funnels, and obtains similar results. By applying ultrasound in cacao seeds, the efficiency of extraction increased by 57.7 % for CF and 43.6 % for TB and by between 12 and 23 % for both analytes in chocolate products, in comparison with those results obtained by a conventional stirring extraction method.

In LC full scan based MS screening methods correct mass assignment is essential. Parameters affecting the accuracy of mass assignment, i.e., analyte concentration, complexity of the matrix, and resolving power, were studied using typical examples from the field of residue and contaminant analysis in food and feed. The evaluation was carried out by analyzing samples of honey and animal feed, spiked with 151 pesticides, veterinary drugs, mycotoxins, and plant toxins at levels ranging from 10 to 250 ng/g. Analyses were performed using a single stage Orbitrap with resolving power settings varying from 10,000 to 100,000 (FWHM). For consistent and reliable mass assignment (<2 ppm) of analytes at low levels in complex matrices, a high resolving power (≥50,000) was found to be required. At lower resolving power settings, the error in the assignment of mass increased due to the coelution of analytes with interferences at the same nominal mass. This negatively affected selectivity and quantitative performance due to the inability to use the required narrow mass-extraction windows. In the case of the less complex honey matrix, a resolving power of 25,000 was generally sufficient to obtain a mass assignment error close to the typical instrument mass accuracy (≤2 ppm) down to low concentration levels of 10 ng/g. Mass resolving power and complexity of the sample are major parameters affecting the accuracy of mass assignment in multi-residue screening applications.

Control of chocolate viscosity is vital to its quality and production cost, and directly influenced by solids particle size distribution (PSD) and composition. Effects of PSD and composition on rheological properties of molten dark chocolate were investigated by varying PSD [D₉₀ (90% finer than this size) of 18, 25, 35 and 50 μm], fat 25, 30 and 35% and lecithin (0.3 and 0.5%) using a shear rate-controlled rheometer. PSD, fat and lecithin content significantly affected all rheological parameters, with significant interaction among factors. Increasing particles size gave significant reductions in Casson plastic viscosity, Casson yield value, yield stress, apparent viscosity and thixotropy, with greatest effect with 25% fat and 0.3% lecithin, which reduced with increasing fat and lecithin contents. Statistical analysis revealed that fat exerts the greatest effect on the variability in all the rheological properties followed by PSD and lecithin. PSD, fat and lecithin could be manipulated to control dark chocolate rheology, influencing quality whilst reducing production cost.

The aim of this investigation was to estimate honeys and bee products quality in view of their mineral composition using multivariate techniques. Fourteen elements (Ca, Mg, K, Na, P, Co, Mn, Fe, Cr, Ni, Zn, Cu, Cd, and Pb) were determined in 66 honeys and bee products from different places of Poland and Europe and various botanical origins. The total metals contents were analyzed by flame atomic absorption spectrometry using deuterium-background correction after wet digestion with nitric acid in an automatic microwave digestion system. Phosphorus was determined in the form of phosphomolybdate by a spectrophotometric method. Reliability of the procedure was checked by analysis of the certified reference materials tea (NCS DC 73351) and cabbage (IAEA-359). The analytical data indicated a good level of quality of honeys, especially with regard to the concentration of toxic trace elements, such as Cd and Pb. Results were submitted to multivariate analysis, including such techniques as factor and cluster analyses in order to evaluate the existence of data patterns and the possibility of classification of honeys from different botanical origins according to their mineral content. The nine metals determined were considered as chemical descriptors of each sample. There was a significant influence of the botanical and geographical provenance as well as technological processing on the elemental composition of honeys.

Melatonin functions as a free radical scavenger and controls the regulation of the sleep-wake cycle in mammals, while serotonin is the main intermediate in melatonin biosynthesis. In this paper, melatonin and serotonin have been detected and quantified for the first time in eight different sweet cherry cultivars using high performance liquid chromatography with mass spectrometry detection. The limits of detection of the proposed analytical method were 4.3 ng/mL for melatonin and 3.0 ng/mL for serotonin. An inverse relation between the contents of melatonin and serotonin was observed in the studied sweet cherry cultivars. The highest melatonin amounts were found in 'Burlat' sweet cherries (22.4 ± 1.3 ng/100 g of fresh fruit), while the highest serotonin contents were found in the cultivar 'Ambrunés' (37.6 ± 1.4 ng/100 g of fresh fruit). The results presented in this research allow us to know the amount of melatonin and serotonin bring to the diet.

The stabilities of three natural blue colorants--gardenia blue, phycocyanin and indigo--toward heat and light were studied in simple solutions of various pH. Gardenia blue was found to be stable at temperatures up to 80 °C in aqueous solution at pH 3, 5 and 7. Exposure to light of 3×10^sup 5^ lux for 24 hours resulted in approximately 50% degradation of gardenia blue in aqueous solution. Phycocyanin was found to be unstable to heat and light in aqueous solution. Phycocyanin is insoluble in acidic solution (pH 3) and denatures at temperatures above 45 °C at pH 5 and 7, leading to a color change. Exposure to light of 3×10^sup 5^ lux for 24 hours in aqueous solution at pH 5 and 7 caused ~80% degradation. Indigo was found to be stable in medium-chain-triglyceride oil for temperatures up to 90 °C, but to photodegrade (by approximately 70% after five hours of exposure to 3×10^sup 5^ lux), corresponding to a quantum yield of 1.8×10^sup -4^ mol Einstein^sup -1^ for visible light and 1.4×10^sup -2^ mol Einstein^sup -1^ for UV, as determined for monochromatic light of 600 and 313 nm, respectively. Exploratory studies have been carried out with the three blue colorants in application media such as soft drink, jelly gum, hard candy and sugar coating for soft candy. Despite its lower stability towards heat and light, phycocyanin was concluded to be the more versatile blue food colorant among the three studied, showing a bright blue color in jelly gum and coated soft candy.[PUBLICATION ABSTRACT]

Purpose An analytical method for quantitation of sibutramine in human hair using gas chromatography (GC)–isotope dilution tandem mass spectrometry (MS/MS) was newly established. In this article, a case is presented, in which a 3.5-year-old male child accidentally ingested chocolate-like product containing sibutramine, showing various symptoms; he could survived the crisis. About 1 month after the incident, his scalp hair sample was subjected to analysis for the causative sibutramine. Method After cryo-grinding for the hair sample, target compound was extracted with methanol, and the solvent layer was evaporated to dryness. The residue was reconstituted in methanol and analyzed by GC–MS/MS, using the selected reaction monitoring (SRM) mode with a deuterated isotope internal standard. Results The substance was identified as sibutramine; its concentration in the hair sample of the child was 58.6 pg/mg. The calibration curve of sibutramine in hair samples had a good linear relationship in the concentration range of 20–200 pg/mg ( r  > 0.99); the extraction recovery rate 85.2–91.8%; the interday and intraday precision and accuracy (bias) examined not greater than 9.6%. Sibutramine in human hair had good stability under 3 different storage conditions at room (20 °C), refrigerated (4 °C) and frozen ( – 20 °C) temperatures for at least 7 days. Conclusions It should be expected that the method established in this study would contribute to rapid determinations of sibutramine. To our knowledge, this is the first report describing quantitation of sibutramine in an authentic human hair sample by GC–MS/MS.