Development and validation of a sensitive and simultaneous liquid chromatography tandem mass spectrometry method for the determination of eight phytocannabinoids in various CBD products.

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.

Rupture Endometriomas Presenting as Acute Abdomen Infection in Hasty and Limited Resources Setting: A Pitfall Not to Miss - A Case Report.

Introduction: Abdominal pain is a diagnostic problem that requires immediate care and treatment for surgeons and gynecologists. The causes of abdominal pain in women of childbearing age range from benign and temporary to potentially life threatening. Rare etiologies such as spontaneous ruptured endometrioma are often not included in the radar of diagnosis due to their rarity and non-specific signs and symptoms in the patient. This case report aimed to show a resemblance between the clinical symptoms of acute abdomen in diffuse peritonitis due to hollow viscus perforation and spontaneous ruptured endometrioma. Case Description: A 42-year-old woman presented to our hospital with abdominal pain. She had a history of fever for two weeks. She came from a tropical rural area where typhoid fever is common. She was advised to undergo emergency laparotomy because of the suspicion of diffuse peritonitis due to a hollow viscus perforation due to typhoid infection. Because of acute abdominal pain, a vertical incision was made to explore her abdominal cavity, and chocolate-like fluid and ovarian cysts were found during surgery. The diagnosis was changed to diffuse peritonitis due to spontaneous rupture of the endometrioma bilaterally. Conclusion: This case suggests that the exact diagnosis and cause of abdominal pain varies. As the current gold standard for endometrioma is laparoscopy, surgeons must prepare a collaborative approach to the cause of the disease.

Abdominal pain is common and often requires prompt medical care. In women, it can have various causes, including rare ones, such as spontaneous ruptured endometrioma, which can be difficult to diagnose owing to its rarity and vague symptoms. We describe the case of a 42-year-old woman with abdominal pain and fever, initially suspected to have diffuse peritonitis due to typhoid infection. However, surgery revealed a ruptured endometrioma, highlighting the challenge of accurately diagnosing such conditions. This case emphasizes the need to consider the diverse diagnoses of abdominal pain and the importance of collaborative approaches among medical professionals to ensure accurate diagnosis and treatment.

Flow properties of single origin chocolates: Effect of product formulation and particle size.

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.

Intake of Dark and Milk Chocolate Differently Affects Autonomic Nervous Control of the Heart During Rest and Mental Stress.

OBJECTIVE: Chocolate is a popular food that may affect the activity of the autonomic nervous system (ANS). The aim of this study was to determine the effect of a single dose of dark or milk chocolate on ANS cardiac control during rest and mental stress induced by the Stroop test (ST). METHODS: Healthy participants, divided into DARK or MILK chocolate groups, ingested corresponding type of chocolate (1 g/kg body weight). They underwent measurement of ANS during relaxation and ST before and 2 h after chocolate consumption. ANS control was assessed by determination of heart rate (HR) and heart rate variability using parameters related to complex autonomic modulation (TP, SDNN) or primary vagal modulation (HFnu, RMSSD). RESULTS: HR was always increased during ST in both groups. Relaxation HR values after chocolate ingestion were higher only in the DARK chocolate group. During ST, values of TP, SDNN and HFnu decreased before and after chocolate ingestion in the DARK group, but only before chocolate ingestion in the MILK group. RMSSD values decreased during ST before and after chocolate ingestion in both groups. Relaxation TP, RMSSD and HFnu values after chocolate ingestion were lower in the DARK but not in the MILK group. CONCLUSION: The results suggest that even a single dose of milk chocolate attenuates changes in ANS cardiac control induced by mental stress, whereas a single dose of dark chocolate has an activating effect on the heart via modification of ANS cardiac control at rest. Different levels of sugars and cocoa biologically active compounds in the two types of chocolate could explain the observed effects.

Dark chocolate: delivery medium for probiotic Bifidobacterium breve NCIM 5671.

Bifidobacteria are widely acclaimed probiotic bacteria, however, the fragile nature of the bacteria has rendered its delivery through food products a challenge. The aim of the present study was to develop probiotic dark chocolate by incorporating Bifidobacterium breve NCIM5671. The probiotic chocolate was prepared by adding B. breve to dark chocolate at the final tempering stage. The chocolate was evaluated for the viability of B. breve upon preparation and during storage period of 90 days. The effect of addition of B. breve on physiological parameters of chocolate such as color, texture, rheology, melting profile, and sensory profile was also determined. The probiotic chocolate developed retained viability of B. breve (9 log CFU/g) for a period of 90 days. No significant differences were observed in physiological parameters of probiotic chocolate compared to control chocolate. Overall the probiotic dark chocolate was found to be a suitable matrix for delivery of B. breve NCIM5671. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-024-05958-6.

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

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

Research on Application of Japanese Quince (Chaenomeles L.) and Pork Collagen in Dark Chocolate-Benefits in Prevention of Inflammation In Vitro Model.

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.

An approach to manufacturing well-being milk chocolate in partial replacement of lecithin by the functional plant-based combination.

Lecithin is constituted of a glycerophospholipid mixture and is abundantly used as an emulsifying agent in various food applications including chocolate production. However, overconsumption of lecithin may create an adverse effect on human health. Thus, this study aims to replace the lecithin with plant-based gums. Different ratios of guar and arabic gum (25%-75%) and their blend (25%-75%) were employed as partial replacement of lecithin. Milk chocolate prepared using 40% guar gum (60GGL [guar gum, lecithin]), 25% arabic gum (75AGL [arabic gum, lecithin]), and a blend of 15 arabic gum and 10 guar gum (65AGGL [arabic gum, guar gum, lecithin]) showed similar rheological behavior as compared to control chocolate (100% lecithin). The fat content of 65AGGL (37.85%) was significantly lower than that of the control sample (43.37%). Rheological behavior exhibited shear-thinning behavior and samples (60GGL-75GGL-80GGL, 65AGL-75AGL, and 65AGGL-75AGGL) showed similar rheological properties as compared to control. The chocolate samples (60GGL and 65AGGL) showed significantly (p < .05) higher hardness values (86.01 and 83.55 N) than the control (79.95 N). As well, gum-added chocolates exhibited higher thermal stability up to 660°C as compared to the control sample. The Fourier transform infrared spectroscopy (FTIR) analysis revealed predominant ß-(1 → 4) and ß-(1 → 6) glycosidic linkages of the gums and lecithin. Sensory evaluation revealed a comparable score of gum-added milk chocolate in comparison to control samples in terms of taste, texture, color, and overall acceptance. Thus, plant exudate gums could be an excellent alternative to lecithin in milk chocolate, which can enhance the textural properties and shelf life.

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

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

Identification of Faba bean genetic loci associated with quantitative resistance to the fungus Botrytis fabae, causal agent of chocolate spot.

Introduction: Chocolate spot, caused by the ascomycete fungus Botrytis fabae, is a devastating foliar disease and a major constraint on the quality and yield of faba beans (Vicia faba). The use of fungicides is the primary strategy for controlling the disease. However, high levels of partial genetic resistance have been identified and can be exploited to mitigate the disease. Methods: The partially resistant V. faba cultivar Maris Bead and susceptible Egyptian accession ig70726 were crossed, and a genetic mapping population of 184 individuals was genotyped in the F2 generation and screened for resistance to B. fabae infection in the F3, F5, and F6 generations in a series of field experiments. A high-density linkage map of V. faba containing 3897 DArT markers spanning 1713.7 cM was constructed. Results: Multiple candidate quantitative trait loci (QTLs) in 11 separate regions of the V. faba genome were identified; some on chromosomes 2, 3, and 6 overlapped with loci previously linked to resistance to Ascochyta leaf and pod blight caused by the necrotrophic fungus Ascochyta fabae. A transcriptomics experiment was conducted at 18 h post-inoculation in seedlings of both parents of the mapping population, identifying several differentially expressed transcripts potentially involved in early stage defence against B. fabae, including cell-wall associated protein kinases, NLR genes, and genes involved in metabolism and response to reactive oxygen species. Discussion: This study identified several novel candidate QTLs in the V. faba genome that contribute to partial resistance to chocolate spot, but differences between growing seasons highlighted the importance of multi-year phenotyping experiments when searching for candidate QTLs for partial resistance.

Optimization of N fertilizer and synergistic intercropping to enhance the productivity advantage of faba bean and effective control of chocolate spot epidemics.

Intercropping of wheat/faba bean is a common practice within the legume-cereal family. However, the benefits of nitrogen (N) fertilizer optimized synergistic intercropping in improving faba bean productivity while controlling the prevalence of chocolate spot disease have not been established. This study conducted continuous field experiments spanning two planting seasons to investigate two key findings. (1) Optimizing N fertilizer application can enhance the productivity of intercropped faba bean. (2) The percentage severity index (PSI) during the period of maximum prevalence rate (Rmax) of faba bean chocolate spot disease, poses a substantial challenge to faba bean yield. The results indicated that the land equivalent ratio and transgressive overyielding index for each intercropping treatment increased with higher N fertilizer application, exceeding a value of 1, and the land saving proportion also exceeded 0. Intercropping primarily enhances productivity, as measured by the harvest index (HI), by amplifying the complementary effect rather than the selection effect, thus improving the net benefit of intercropping. The HI of single and intercropped faba bean increased with the N1 and N2 treatments in both planting seasons. However, the HI of single and intercropped faba bean at the N3 level decreased significantly, ranging from 17.85% to 29.62%. Furthermore, a notable negative correlation was established between the PSI during critical epidemic (initial epidemic, maximum epidemic rate, and late epidemic) periods and observed and expected faba bean yields. As PSI increased, faba bean yields decreased and PSI of intercropping at different periods were lower than those observed in the single cropping. Additionally, intercropping with the optimized N fertilizer treatment (N2 treatment) exhibited an enhanced relative control effect on chocolate spot disease in faba bean, ranging from 35.21% to 52.36%. This finding confirmed the productivity advantage of intercropping faba bean. In conclusion, this study suggested that optimizing N fertilizer application can enhance the productivity of intercropped faba bean. Wheat/faba bean intercropping effectively controlled the PSI during the period of Rmax, which would otherwise threaten faba bean yield. Consequently, this practice ensured sustained advantages of wheat/faba bean intercropping.

Occurrence and Synthesis Pathways of (Suspected) Genotoxic α,ß-Unsaturated Carbonyls in Chocolate and Other Commercial Sweet Snacks.

α,ß-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,ß-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,ß-unsaturated carbonyls, while wet conching partially restored or increased their levels due to cocoa butter addition. Further analyses showed that α,ß-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,ß-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).

Shades of Blue: A Case Series of Acquired Methemoglobinemia.

Acquired methemoglobinemia (MetHb) is a rare but potentially life-threatening condition that has varied etiology, usually toxin- or drug-induced. We had five cases of acquired methemoglobinemia during six months. Their presentation varied from an asymptomatic state to respiratory distress. The presence of cyanosis and low oxygen saturation (SpO2), despite normal partial pressure of oxygen (PaO2) and chocolate brown-colored blood, were diagnostic clues present in all cases. A high level of methemoglobinemia was detected on arterial blood gas (ABG), confirming the diagnosis. Methylene blue was used as an antidote along with supportive care in symptomatic cases. All these cases of methemoglobinemia recovered completely. A high index of suspicion for methemoglobinemia should be maintained in cases presenting with persistent hypoxia or cyanosis despite normal PaO2.

Immune Dysregulation in Endometriomas: Implications for Inflammation.

The most common manifestation of endometriosis, a condition characterized by the presence of endometrial-like tissue outside of the uterus, is the endometrioma, a cystic ovarian lesion. It is a commonly occurring condition associated with chronic pelvic pain exacerbated prior to and during menstruation, as well as infertility. The exact pathomechanisms of the endometrioma are still not fully understood. Emerging evidence suggests a pivotal role of immune dysregulation in the pathogenesis of endometriomas, primarily influencing both local and systemic inflammatory processes. Among the factors implicated in the creation of the inflammatory milieu associated with endometriomas, alterations in both serum and local levels of several cytokines stand out, including IL-6, IL-8, and IL-1ß, along with abnormalities in the innate immune system. While numerous signaling pathways have been suggested to play a role in the inflammatory process linked to endometriomas, only NF-κB has been conclusively demonstrated to be involved. Additionally, increased oxidative stress, both resulting from and contributing to endometriomas, has been identified as a primary driver of both systemic and local inflammation associated with the condition. This article reviews the current understanding of immune dysfunctions in the endometrioma and their implications for inflammation.

Effectiveness of Dark Chocolate and Carrot Juice on Perceived Labor Pain During Stage 1 of Birth Delivery Among Primigravida: A Quasi-Experimental Study.

Background: Dark chocolate and carrot juice may positively decline the pain. However, there is a lack evidence the impact of combination dark chocolate and carrot juice on labor pain during stage 1 of birth delivery among primigravida. Objective: This study aimed to examine the effectiveness of dark chocolate and carrot juice on perceived labor pain during stage 1 of birth delivery among primigravida. Methods: This was a quasi-experimental study with participants who received dark chocolate (n=30), carrot juice (n=30), and control group (n=30). Pain level was assessed by using the Numeric Pain Rating Scale (NPRS) before the intervention and at 30 hours after intervention. The Chi-square and one-way analysis of variance tests and general equational model were used. Results: Data were collected and analyzed before and after 60 minutes of intervention. Our results showed a significant interaction between the group and time, with both groups independently ameliorating labor pain. Conclusion: Dark chocolate and carrot juice therapies independently lowered pain labor in primigravida mothers, making them a viable treatment for advanced pain labor.

Assessing agronomic performance, chocolate spot resistance, and heat tolerance for diverse Vicia faba genotypes under varying environmental conditions.

Chocolate spot and heat stress devastatingly impact the production of faba bean, particularly under prevailing climatic changes and rising drastic environmental conditions. Hence, the adaptability of faba bean performance is a decisive objective of plant breeders to ensure its sustainable production. The present study aimed to evaluate the agronomic performance and stability of diverse eleven faba bean genotypes for yield characters, chocolate spot, and heat stress in eight different growing environments. The faba bean genotypes were evaluated at two sowing dates in two different locations during two growing seasons. The evaluated eleven faba bean genotypes were sown timely in autumn (25 October) and late sowing in early winter (25 November) in Bilbeis and Elkhatara during 2020 and 2021 growing seasons. The results exhibited substantial differences among the evaluated sowing dates, locations, and faba bean genotypes for all studied characters. The genotypes Sakha-3, Nubaria-3, Nubaria-5, Misr-3, and Wadi-1 were able to produce acceptable yield and quality characters under timely sowing in autumn and late sowing in early winter in all tested environments. Moreover, the genotypes Nubaria-3, Nubaria-4, Nubaria-5, Sakha-4, Giza-3, and Triple White exhibited better resistance to chocolate spot. The assessed faba bean genotypes were evaluated under late sowing to expose the plants to high temperature stress at flowering and throughout the anthesis and seed-filling stages. The genotypes Nubaria-5, Nubaria-3, Nubaria-4, Sakha-3, Sakha-4, Wadi-1, and Misr-3 possessed tolerance to heat stress more than the other genotypes. Different statistical methods were applied to study the stability of assessed genotypes such as joint regression, Additive Main Effect and Multiplicative Interaction (AMMI) analysis, AMMI stability value, Wricke's and Ecovalence values. The estimated stability parameters were consistent in depicting the stability of the assessed faba bean genotypes. The findings revealed that Sakha-1, Misr-3, Nubaria-4, and Nubaria-5 demonstrated stable and desirable performance across all tested environments. The heatmap was employed to classify the assessed faba bean genotypes into different groups based on agronomic performance, chocolate spot resistance and heat stress tolerance. Nubaria-3, Nubaria-4, Nubaria-5, and Misr-3 had the best performance for agronomic performance, chocolate spot resistance, and heat stress tolerance. The obtained results provide evidence of employing promising faba bean genotypes for improving the stability of agronomic performance, chocolate spot resistance, and heat stress tolerance in breeding programs principally under unprecedented climate fluctuations.

Applications of Life Cycle Assessment in the Chocolate Industry: A State-of-the-Art Analysis Based on Systematic Review.

Chocolate is a popular food for its unique flavor and taste, rich nutritional value, and the psychological values brought to people. The raw material production of chocolate, product manufacturing, sales and transportation have different degrees of environmental impact. This review explores the environmental hot spots in the life cycle of chocolate and puts forward corresponding suggestions for the improvement. By applying a systematic review method, this paper collected 25 articles on life cycle assessment (LCA) of the environmental impact of the chocolate industry. It is found that the life cycle of chocolate has the highest environmental impact in the raw material production and chocolate manufacturing stages (accounting for 77-97% of total impacts), among which milk powder, sugar and cocoa derivatives are the important contributors to significant environmental burden. Dark chocolate generates the lowest carbon emissions (1.67 kg CO2 eq/kg product) among existing chocolate categories, while the chocolate confectionery products release the highest carbon emissions (6.76 kg CO2 eq/kg product) among chocolate-containing products. Improvement measures are proposed for reducing environmental impacts and for selecting environmentally friendly product formulae. This study can provide benchmarking for the chocolate industry and improves the understanding of life cycle environmental impacts of chocolate products.

Counts of mesophilic aerobic, mesophilic anaerobic, thermophilic aerobic sporeforming bacteria and persistence of Bacillus cereus spores throughout cocoa powder processing chain.

Sporeforming bacteria are a concern in some food raw materials, such as cocoa powder. Samples (n = 618) were collected on two farms and at several stages during cocoa powder manufacture in three commercial processing lines to determine the impact of each stage on bacterial spore populations. Mesophilic aerobic, mesophilic anaerobic, thermophilic aerobic, and Bacillus cereus spore populations were enumerated in all the samples. Genetic diversity in B. cereus strains (n = 110) isolated from the samples was examined by M13 sequence-based PCR typing, partial sequencing of the panC gene, and the presence/absence of ces and cspA genes. The counts of different groups of sporeforming bacteria varied amongst farms and processing lines. For example, the counts of mesophilic aerobic spore-forming (MAS) populations of cocoa bean fermentation were lower than 1 log spore/g in Farm 1 but higher than 4 log spore/g in Farm 2. B. cereus isolated from cocoa powder was also recovered from cocoa beans, nibs, and samples after roasting, refining, and pressing, which indicated that B. cereus spores persist throughout cocoa processing. Phylogenetic group IV was the most frequent (73%), along with processing. Strains from phylogenetic group III (14 %) did not show the ces gene's presence.

Influence of process conditions of alkalization on quality of cocoa powder.

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.

Choc-Tadalafil Fusion: Unlocking Solubility and Taste Harmony with ß-CD-Infused Medicated Chocolate.

OBJECTIVE: The primary limitations of tadalafil in treating erectile dysfunction are its low solubility and unpleasant bitter taste, which ultimately result in inadequate patient adherence. The present study aimed to develop and characterize a medicated chocolate formulation containing Tadalafil and ß-CD (solubility enhancer) employing the concept of Design of Experiment (DoE) using chocolate as a user-friendly excipient. METHODS: An inclusion complex was formulated by incorporating the drug into ß-CD using the kneading method for solubility improvement and also as a taste masker for Tadalafil. The ratio of drug: ß-CD inclusion complex was selected based on a phase solubility study. The inclusion complex was molded into a chocolate base and optimized using the DoE approach. Further, drug excipient interaction was evaluated by DSC and FTIR study. RESULT: Phase solubility study suggested a 1:1 ratio of Tadalafil: ß-CD for better solubility. DSC spectra suggested the conversion of crystalline structure into an amorphous state which indicates improvement of the drug solubility. DSC and FTIR studies revealed that there was no significant interaction between drug and excipients. Next, %CDR (cumulative drug release) at 30 min revealed the immediate effect of Tadalafil from chocolate formulation and free drug analysis (an unbound drug with ß-CD) proved reduced bitterness of the drug in the complex. Additionally, the medicated chocolate was found to be stable at room temperature as per stability study. CONCLUSION: ß-CD was found to be a promising multifunctional excipient as a solubility enhancement carrier and taste masker for bitter-tasting drugs.