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Caffeine and Theobromine Levels in Cocoa and Carob Products

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Abstract

Seventy-nine cocoa or chocolate products and eighteen carob products were analyzed by HPLC for caffeine and theobromine content. After extraction into boiling water, the methylxanthines were identified and quantified with the use of a reverse phase column. Mean theobromine and caffeine levels respectively, were 0.695 mg/g and 0.071 mg/g in cocoa cereals; 1.47 mg/g and 0.152 mg/g in chocolate bakery products; 1.95 mg/g and 0.138 mg/g in chocolate toppings; 2.66 mg/g and 0.208 mg/g in cocoa beverages; 0.621 mg/g and 0.032 mg/g in chocolate ice creams; 0.226 mg/g and 0.011 mg/g in chocolate milks; 74.8 mg/serving and 6.5 mg/serving in chocolate puddings. Theobromine and caffeine levels in carob products ranged from 0-0.504 mg/g and 0-0.067 mg/g, respectively.

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... Furthermore, processing of cocoa beans affects the concentrations of methylxanthines and their content in cocoa products may significantly vary compared to cocoa beans [12,101,102]. In contrast to cocoa, caffeine and theobromine are either not detected or present in trace amounts in carob powder and carob products [32,103,104]. Hence, substitution of cocoa powder by carob powder is suggested as an effective approach to decrease caffeine and theobromine to trace levels in confectioneries, beverages and baking products [103,104]. ...
... In contrast to cocoa, caffeine and theobromine are either not detected or present in trace amounts in carob powder and carob products [32,103,104]. Hence, substitution of cocoa powder by carob powder is suggested as an effective approach to decrease caffeine and theobromine to trace levels in confectioneries, beverages and baking products [103,104]. Theophylline has been detected in the carob pulp extracts of certain carob cultivars, while it is absent in others [33]. The presence of theophylline should be further investigated in additional carob cultivars as well as in carob products. ...
Article
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Cocoa originates from the beans of the cocoa tree (Theobroma cacao L.). It is an important commodity and the main ingredient in chocolate manufacture. Its value and quality are related to complex flavors and to its distinct sensory properties. The increasing demand for cocoa and its rising price urges the research for cocoa substitutes. A potential substitute for cocoa is carob. Carob is the fruit of an evergreen tree (Ceratonia siliqua L.) cultivated in the Mediterranean area, well known for its valuable locust bean gum and also for carob powder and syrup that are obtained from carob pulp. Cocoa beans and carob pods contain various phytochemicals including polyphenols, proteins and amino acids, fatty acids, carbohydrates and fiber. Phytochemicals represent an important source of nutrients and compounds that are beneficial to human health. In this review, phytochemicals in cocoa beans and carob pods and their impact on human health are reviewed. The bioactive compounds that are present in carob, in conjunction with the cocoa-like flavors and unique sensory properties that are enhanced by carob powder roasting, underline carob’s potential to substitute cocoa in various food products. These food applications are discussed in this review.
... In comparison with cocoa powder, the absence of caffeine was observed in the carob powder prepared for this study, as proved by Craig and Nguyen [63]. Such result could be considered as an advantage for the use of carob powder as a cocoa powder substitute for the production of caffeine-free food products. ...
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The by-product generated from carob molasses processing is considered as an excellent source of dietary fiber and may be used as a functional ingredient in food industry. However, it presents a high value of water activity (~ 0.98) which facilitates its microbiological contamination and rapid deterioration. So that, this study provides a solution for the valorization of this by-product and suggests the incorporation of the dried carob by-product into Halva to produce an added value product (Halva with carob powder). Thus, the present work focused on the characterization of carob powder and the optimisation of incorporation percentage of carob powder into Halva formulation. The characterization showed the absence of caffein in carob powder compared to cocoa’s one. Besides, carob and cocoa powders had both a brown color. The former had lower fat and higher sugar contents compared to the latter. The optimization promoted the addition of 5% carob powder into Halva formulation according to the evaluation of hardness, sensory quality and exudative stability. Therefore, the new confectionary product could be considered as a promising nutritious and healthy foodstuff to consumers.
... TB is an alkaloid molecule in the xanthine family that occurs in the cocoa 'bean', and dark chocolate consists of about 1 to 4% TB [14]. TB is related to caffeine and theophylline, but it has weaker effects on the central nervous system than caffeine [15]. ...
Article
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Purpose Uric acid renal lithiasis has a high prevalence and a high rate of recurrence. Removal of uric acid stones can be achieved by several surgical techniques (extracorporeal shock wave lithotripsy, endoscopy, laparoscopy, open surgery). These stones can also be eliminated by dissolution within the kidneys, because the solubility of uric acid is much greater when the pH is above 6. At present, N-acetylcysteine with a urinary basifying agent is the only treatment proposed to increase the dissolution of uric acid stones. In this paper, we compare the effect of theobromine and N-acetylcysteine on the in vitro dissolution of uric acid calculi in artificial urine at pH 6.5. Methods The dissolution of uric acid renal calculi was performed in a temperature-controlled (37 °C) chamber. A peristaltic pump was used to pass 750 mL of synthetic urine (pH 6.5) through a capsule every 24 h. Stone dissolution was evaluated by measuring the change in weight before and after each experiment. Results N-acetylcysteine increased the dissolution of uric acid calculi, but the effect was not statistically significant. Theobromine significantly increased the dissolution of uric acid calculi. Both substances together had the same effect as theobromine alone. The addition of theobromine to a basifying therapy that uses citrate and/or bicarbonate is a potential new strategy for the oral chemolysis of uric acid stones. Conclusion Theobromine may prevent the formation of new stones and increase the dissolution of existing stones.
... Moreover, today's increasing demand is expected to be enhanced even more due to five key factors ( Fig. 33.1 Carob-dried pods can be ground into a powder that resembles the flavor of cocoa. It is commonly used as an alternative to chocolate in health-food products but has advantages over chocolate in that it has fewer calories, and neither caffeine nor theobromine, and oxalic acid (a potential source of kidney stone formation) (Craig & Nguyen, 1984;Yousif & Alghzawi, 2000). Moreover, carob appears to fulfill the modern health criteria of consumers such as gluten-and caffeine-free products, natural chocolate-like sweeteners, ingredients for bread, beverages such as liqueurs, coffee (Papaefstathiou et al., 2018), vegan, rich source of potassium and calcium, and with weight management benefits. ...
Chapter
Ceratonia siliqua L. is a typical Mediterranean tree species that has also been introduced to the temperate regions of Central America, Australia, and Africa. The carob fruit consists of the pulp and the seed (endosperm and germ), each one of which is used in a great variety of bakery products and beverages, as animal feed, food additives, syrups, ice creams, dietetic products, and for the photographic emulsion. For instance, the carob pulp is used to make carob syrup and molasses through the recovery of sugars, whereas the seed is used to produce Locust Bean Gum (LBG), which is used as a growth medium, and as a thickener, chemically known as E410. Currently, there is a strong interest in carob re-cultivation due to the tree’s ability to withstand drought and adapt to climate change, further to its medical and food exploitation potential. In 2016 and 2017, the global carob production exceeded 170,000 tons, with the highest volumes of carob production coming from Spain, Portugal, and Italy, followed by Morocco, Turkey, Greece, Cyprus, Lebanon, and Algeria. Carob fruit harvesting and processing does not produce significant amounts of agricultural and industrial waste. Agricultural waste mainly includes traps for rodents, plastic packaging waste, and field residues such as stems and leaves. On the other hand, industrial processing from carob mills and the food industry produces waste such as solid residual parts and plant biomass (e.g., leaves), which can be composted or used as animal feed. In carob factories, during the roasting and milling processes, air pollutants such as particulate matter (PM) and biogenic volatile organic compounds (VOCs) are released. Carobs and their waste have been studied by various research groups as a source of phenolic compounds and carbohydrates, as an alternate source for the production of biofuels (e.g., biohydrogen and bioethanol), in food packaging applications, in wastewater treatment, as well as a soil organic amendment. Overall, carob and carob waste have promising applications as raw material for further investigation and utilization.Keywords Ceratonia siliqua Locust Bean GumE410Volatile organic compoundsFunctional food
... We recently demonstrated that dietary theobromine is a potent inhibitor of UA crystallization in humans [13,14]. Theobromine is abundant in chocolate and cocoa [15]. Chemically, it is known as 3, 7-dimethylxanthine and, along with caffeine and theophylline, is in the xanthine family of alkaloids. ...
Article
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Background: Uric acid (UA) renal lithiasis has a high rate of recurrence and a prevalence ranging from 10% and 15%, depending on the population. The most important etiological factor is persistence of urinary pH below 5.5 and one of the most common treatments is alkalization with citrate. Recent studies demonstrated that theobromine, which is abundant in chocolate and cocoa, is a potent inhibitor of UA crystallization. Aim: The aim was to compare the efficacy of citrate versus citrate + theobromine as treatment for UA lithiasis. Methods: This randomized cross-over trial investigated the efficacy of two treatments in 47 patients with UA renal lithiasis. Urine volume, pH, UA excretion, theobromine excretion, and risk of UA crystallization (RUAC) at baseline and at the end of each intervention period were measured. Results: Each treatment significantly reduced the risk of UA crystallization compared to basal values. The RUAC after citrate + theobromine was lower than the RUAC after citrate, although this difference was not statistically significant. Conclusion: The combined consumption of citrate and theobromine may be a promising strategy for the prevention of UA kidney stones.
... Several authors have mentioned the possibility of obtaining a chocolate-like flavor from carob powder Lannes 2009, 2010). Craig and Nguyen (1984) asserted that carob is almost free of caffeine and theobromine. This nutrient profile and organoleptic characteristics make carob a potential cacao alternative. ...
Article
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This study deals with the manufacture of a specific cocoa-like powder from Egyptian carob. The crucial unit operation consists of texturing/roasting unseeded carob kibbles, with the aim being to ensure perfect control of the chemical transformations that generate the desired flavor, drying to remove excess moisture, deodorizing to eliminate the undesirable flavor, and texturing to favor the physical/micro-mechanical changes that regulate the microstructure of the end product, in order to obtain an expanded-granule powder that can be used in many food formulations. The effect of saturated steam pressure of 0.30-0.60 MPa (133.5-158.8°C) and processing time of 40-70 s, on the sensory attributes, physical and functional properties, and color characteristics were evaluated and optimized using response surface methodology (RSM). DIC-texturing/roasting could be used to improve the sensory and color characteristics, increase the expansion ratio and oil holding capacity, and decrease the bulk density of DIC-textured/roasted carob powder. The desired quality of textured/roasted carob powder was obtained at 0.60 MPa (158.8°C) for 55 s (experimental conditions) with water holding capacity close to those of non-textured/unroasted sample. DIC can be used as a coupled process of texturing and roasting at 0.60 MPa for 68 s as predicted by RSM.
... Several authors have mentioned the possibility of obtaining a chocolate-like flavor from carob powder Lannes 2009, 2010). Craig and Nguyen (1984) asserted that carob is almost free of caffeine and theobromine. This nutrient profile and organoleptic characteristics make carob a potential cacao alternative. ...
Article
Full-text available
This study deals with the manufacture of a specific cocoa-like powder from Egyptian carob. The crucial unit operation consists of texturing/roasting unseeded carob kibbles, with the aim being to ensure perfect control of the chemical transformations that generate the desired flavor, drying to remove excess moisture, deodorizing to eliminate the undesirable flavor, and texturing to favor the physical/micro-mechanical changes that regulate the microstructure of the end product, in order to obtain an expanded-granule powder that can be used in many food formulations. The effects of saturated steam pressure of 0.30-0.60 MPa (133.5 to 158.8 °C) and processing time of 40-70 s, on the sensory attributes, physical and functional properties, and color characteristics were evaluated and optimized using response surface methodology (RSM). DIC-texturing/roasting could be used to improve the sensory and color characteristics, increase the expansion ratio and oil holding capacity, and decrease the bulk density of DIC-textured/roasted carob powder. The desired quality of textured/roasted carob powder was obtained at 0.60 MPa (158.8 °C) for 55 s (experimental conditions) with water holding capacity close to those of non-textured/unroasted sample. DIC can be used as a coupled process of texturing and roasting at 0.60 MPa for 68 s as predicted by RSM.
... Carob tree extended tree evergreen shadows reach a height of between 15 and 17 meters. At the age of 18, the leg thickness is 85 cm (Craig and Nguyen 1984). Leaves are composed of a long neck with 6-10 opposite leaves, a leaf shape. ...
Chapter
Tropical fruit trees establish critical organic assets in the worldwide agrobiodiversity setting. They differ from the tropical fruit trees of American and Asian source, indigenous fruit tree (IFT) of tropical Africa has barely accomplished the status of the universal acknowledgment in ware markets and research field outside Africa. Indigenous fruits comprise a critical piece of human diets in numerous African countries, especially in provincial regions and during droughts. It is of prime importance to recognize their composition and nutritional values as this will promote and increase the utilization of these fruits. These fruits contain offer high contents of energy, vitamins and minerals, but production and utilization of intriguing and indigenous fruits in Africa are compelled by the absence of buyer mindfulness on their healthy benefits, loss of agro-biodiversity, inefficient or non-existing fruit tree nursery systems, absence of fruit processing facilities and ineffectively developed marketing pathways. Although the IFT can contribute to both, food and nutrition security and also to employment and income generation of rural communities, yet, their capabilities, nonetheless, must be completely abused after taming of these species, which includes recognizable proof and choice of unrivaled mother trees pursued by the spread of the best materials and feasible on-cultivate development of the species. There are around 50–60,000 plant species in the plant kingdom and roughly 3000 of these are utilized in agroforestry ranches. Investigating the huge plant kingdom for trees with potential uses for timber, nourishment, organic product, fuel, fiber, grub and different uses is an exceptional and an energizing test during this period of fast normal asset exhaustion.
... Theobromine is a dimethylxanthine that is abundant in cocoa and cocoa-derived products, such as chocolate [22]. Clinical studies indicated that its half-life in serum is 6.1 to 10 h, and that 16 to 18% of a single dose of 10 mg/kg is excreted unchanged after 48 h [23,24]. ...
Article
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The purpose of this study was to determine the effects of consumption of different cocoa-derived products on uric acid crystallization in urine of 20 healthy volunteers. Participants were requested to select the specific diet that they wished to follow during the 12 h prior to collection of urine. The only restriction was that the diet could not include any product with cocoa, coffee, or caffeine. On the first day, each volunteer followed their selected diet, and an overnight 12 h urine sample was collected as the baseline urine. After seven days on an unrestricted diet, each volunteer repeated the same diet with 20 g of milk chocolate, chocolate powder, or dark chocolate during breakfast and another 20 g during dinner. Overnight 12 h urine samples were then collected. Urine volume, pH, oxalate, creatinine, uric acid, theobromine, and a uric acid crystallization test were determined for each sample. The results for all 20 patients show that uric acid crystallization was significantly lower following the consumption of chocolate powder or dark chocolate relative to baseline or following the consumption of milk chocolate. The results indicated that increased concentrations of urinary theobromine reduced the risk of uric acid crystallization.
... This is in agreement with the results obtained by other researches (Craig & Nguyen, 1984;Kumazawa et al., 2002 The energy value ranges from 280.17 to 286.07 kcal/100 g in carob pods and from 280.40 to 587.59 kcal/100 g in carob products. ...
Article
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Twenty traditional carob products were measured for their nutritional composition, and their results were compared with the pulp of Cypriot carob cultivars. Moisture, ash, fat, proteins, sugars, dietary fibers, minerals, caffeine‐theobromine, carbohydrates, and energy value were determined. Fluctuations of the nutritional composition values based on the ingredients’ chemical synthesis and product manufacturing process were noted. Only 60% of the products had a label indicating their nutritional value, and the majority of them (75%) were consistent with that of labeling. Chemometric analyses distinguished the carob products according to their type and the discriminator components highlighted their particular nutritional value. Carobs can be characterized as functional foods with low‐fat content, high content in dietary fibers, and high content and/or source of minerals; however, carob products partially satisfied those health and nutritional claims as expected. This pilot research contributes to the nutritional estimation of carob and highlights the traditional carob products. Carob pods and their products were analysed for their nutritional composition. Carobs can be characterized as functional foods (fat, dietary fibers and minerals). Their nutritional value was highlighted using Chemometrics
... La farine élaborée à partir de la pulpe (figure 21) peut être utilisée comme ingrédient dans certains aliments, tels que les gâteaux, bonbons, crèmes glacées, boissons (NAS, 1979). De plus, elle est utilisée comme substituant du cacao dans la production du chocolat (figure 21), car elle est moins calorifique et ne contient ni caféine ni théobromine (Craig et Nguyen, 1984). En Egypte, les sirops élaborés à partir de la caroube constituent une boisson populaire (Batlle et Tous, 1997). ...
... Apart polyphenols, cocoa is also rich in methylxanthines (caffeine, theobromine and theophylline) (Jalil & Ismail, 2008; Meng et al., 2009). Theobromine (3,7-dimethylxanthine) is a metabolite of caffeine and is the major methylxanthine in cocoa (Theobroma cacao), constituting about 4% of fat free basis, while caffeine only 0.2% (Jalil & Ismail, 2008; Craig & Nguyen, 1984). Dark chocolate can contain 237-519 mg of theobromine per 50 g portion (Meng et al., 2009). ...
Research
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Polyphenols, principally catechin and epicatechin, as well as methylxanthines (caffeine, theobromine and theophylline) are the two main bioactive compounds found in cocoa. Theobromine is the most abundant methylxanthine present in cocoa and represents 1-4% of non-fat basis. It is a psychoactive compound without diuretic effects and its principal benefits are antioxidant activity, reduction of lipid profile and could have positive effect on cancer. Negative effects of theobromine, principally carcinogenicity, are not well assessed. Majority of studies underline the effects of polyphenols but only a few of them test methylxanthine activity and bioavailability in chocolate matrices. Moreover, synergic interaction between polyphenols and methylxanthine are unclear and need further research. The aims of this study are to first quantify theobromine in different types of chocolate with LC-MS/MS method and to determine bioavailability of theobromine in chocolate samples after in vitro digestion and using Caco-2 cells as model. This approach allows to underline a potential matrix effect due principally to milk proteins in milk chocolate. Moreover effect of polyphenols on theobromine bioavailability is assessing using a white chocolate model composed of theobromine and a mix of polyphenols. Finally, cytotoxicity of different concentrations of pure theobromine on HEp2 cells is assessed. All these information can help for the determination of the toxicity of theobromine in human cells and for the understanding of polyphenols-methylxanthines interaction and the impact on bioavailability.
... So, caffeine content could not be correlated with the products' origin i.e. whether they are imported or domestic. Interestingly, the presence of CF in the chocolate milks observed in this study is similar to the average content of 11 mg/Kg collected by Craig and Nguyen (1984). However, though the CF content found is low, the CF present in milk products may contribute to the exceeding of tolerance limit along with other CF rich products like tea, coffee, energy drinks etc. Again, 7 out of 41 analyzed dairy products were found BA positive. ...
... So, caffeine content could not be correlated with the products' origin i.e. whether they are imported or domestic. Interestingly, the presence of CF in the chocolate milks observed in this study is similar to the average content of 11 mg/Kg collected by Craig and Nguyen (1984). However, though the CF content found is low, the CF present in milk products may contribute to the exceeding of tolerance limit along with other CF rich products like tea, coffee, energy drinks etc. Again, 7 out of 41 analyzed dairy products were found BA positive. ...
Article
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Background The aim of this current study was to investigate the presence and to determine the contents of caffeine, preservatives and antioxidants in dairy products available in Bangladesh, one of the alarming countries in terms of food security. As high as 41 marketed dairy products, of both locally and internationally manufactured, were collected from local markets of Dhaka, Bangladesh in mid–2013 and then analyzed using an RP–HPLC method to determine the contents of caffeine, benzoic acid, propylparaben, butylparaben, butylated hydroxyanisole and butylated hydroxytoluene. The evaluation was performed using a C18 column (150 mm × 4.6 mm i.d., 5 μm particle size) with a gradient flow rate of acetonitrile and diluted sulfuric acid (0.002 M) as mobile phase from ratio 15:85 to 80:20 (%v/v) at a flow rate of 2.0 ml/min at 265 nm. ResultsThe retention times of CF, BA, PP, BP, BHA and BHT were found about 3.6, 11.6, 13.0, 13.3, 13.6 and 16.7 min, respectively. Results revealed that BA was found in 17.1 % of the products within the concentration range of 11 ~ 2067 mg/L; among these, 42.9 % products exceeded the allowable limit of 300 mg/L set by JECFA. Moreover, 14.6 % products showed positive response to CF at a concentration of 10 ~ 18 mg/L, which was well below the tolerance limit (<200 mg/L) set by FDA. However, none of these investigated dairy products were found to contain any detectable amount of PP, BP, BHA or BHT. Conclusion Presence of excess amount of BA in dairy products, which are one of the most favorite healthy food items for all generations, can easily jeopardize public health sector. Moreover, CF was found in some CF–free products, which is also an alarming issue to be considered seriously to prevent the future occurrence of it.
... Carob kibble is high in sugar content which ranges from 30% to 60% with the main sugars being sucrose (65% to 75% of the total sugars), fructose and glucose (15% to 25% of the total sugars) (Ayaz Lannes 2009, 2010). Unlike chocolate or cocoa, it is almost free from caffeine and theobromine (stimulants) and oxalic acid (a potential source of kidney stone formation) (Craig and Nguyen 1984). Carob kibble also contains very low fat (about 1%) and sodium contents making it a healthy food ingredient (Makris and Kefalas 2004). ...
Article
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Carob (Ceratonia siliqua L.) is well known for its valuable locust bean gum obtained from the carob seeds. Separation of seeds from the pod leaves behind the carob kibble which is a good source of dietary fiber, sugars, and a range of bioactive compounds such as polyphenols and pinitol. Bioactive compounds present in carob kibble have been found to be beneficial in the control of many health problems such as diabetes, heart diseases, and colon cancer due to their antidiabetic, antioxidant, and anti-inflammatory activities. Carob kibble has substantial potential to be used as a food ingredient. This article focuses on the composition, health benefits, and food applications of carob kibble.
... Carob is often used as a chocolate or cocoa substitute (Brand, 1984; Nyerges, 1978). Such usage is attributed to the fact that carob has the advantage of being caffeine and theobromine-free whereas chocolate and cocoa contain relatively high amounts of these two antinutrients (Craig and Nguyen, 1984). Carob contains about 18% cellulose and hemicellulose. ...
Article
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CSpreadable creams are solid-oil suspensions, a mix of fats represents the oil phase, the dispersed phase consisting usually of sugar, cocoa powder, milled and roasted nuts, dried milk and whey. For improving the viscosity of the final product emulsifiers are used, most common being lecithin and mono and diglycerides. The present paper refers to a spreadable confectionery product group, creamy, proper to be eaten as it is, as well as spread on a bread slice or as a filling for cookies or chocolate cream. According to this work, the following ingredients were used: roasted sunflower kernels, sugar, palm oil, cocoa or carob powder and lecithin. The obtained product can be consumed also by persons who suffer from allergies, due the fact that sunflower seeds were replacing the peanuts or almond, the ingredients known as allergens and which are usually used in the technological process of obtaining these creams. The purpose of this study was to characterize the obtaining confectionery spreadable creams based on sunflower kernels, cocoa or carob powder. It was determined the chemical composition of the prototypes obtained, a spreadable cream having no cocoa or carob, one with cocoa and one with carob powder. The antioxidant capacity and total phenolic content of the obtained samples were also assessed.
... Due to the lack of uric acid crystallization inhibitors, the treatment of patients prone to the formation of uric acid stones is based on urine alkalinization, and the administration of allopurinol to patients with hyperuricemia. Theobromine is a dimethylxanthine present in high amounts in chocolate and cocoa [28]. Theobromine has been less well studied than other natural methylxanthines (figure 1) because it stimulates the central nervous system in a lesser degree [29]. ...
Article
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Purpose To assess the capacity of methylxanthines (caffeine, theophylline, theobromine and paraxanthine) to inhibit uric acid crystallization, and to evaluate their potential application in the treatment of uric acid nephrolithiasis. Materials and Methods The ability of methylxathines to inhibit uric acid nucleation was assayed turbidimetrically. Crystal morphology and its modification due to the effect of theobromine were evaluated by scanning electron microscopy (SEM). The ability of theobromine to inhibit uric acid crystal growth on calculi fragments resulting from extracorporeal shock wave lithotripsy (ESWL) was evaluated using a flow system. Results The turbidimetric assay showed that among the studied methylxanthines, theobromine could markedly inhibit uric acid nucleation. SEM images showed that the presence of theobromine resulted in thinner uric acid crystals. Furthermore, in a flow system theobromine blocked the regrowth of post-ESWL uric acid calculi fragments. Conclusions Theobromine, a natural dimethylxanthine present in high amounts in cocoa, acts as an inhibitor of nucleation and crystal growth of uric acid. Therefore, theobromine may be clinically useful in the treatment of uric acid nephrolithiasis.
... Higher-quality chocolate tends to contain more theobromine than lower-quality chocolate. The mean theobromine content of cocoa beans is approximately 20.3 mg/g [Craig and Nguyen, 1984]. ...
Article
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Objective: This study investigated the remineralization potential of theobromine in comparison to a standard NaF dentifrice. Methods: Three tooth blocks were produced from each of 30 teeth. Caries-like lesion was created on each block using acidified gel. A smaller block was cut from each block for baseline scanning electron microscopy imaging and electron-dispersive spectroscopy (EDS) analysis for surface Ca level. A tooth slice was cut from each lesion-bearing block for transverse microradiography (TMR) quantification of baseline mineral loss (Δz) and lesion depth (LD). Then baseline surface microhardness (SMH) of each lesion was measured. The three blocks from each tooth were assigned to three remineralizing agents: (1) artificial saliva; (2) artificial saliva with theobromine (0.0011 mol/l), and (3) NaF toothpaste slurry (0.0789 mol/l F). Remineralization was conducted using a pH cycling model with storage in artificial saliva. After a 28-day cycle, samples were analyzed using EDS, TMR, and SMH. Intragroup comparison of pre- and posttest data was performed using t tests (p < 0.05). Intergroup comparisons were performed by post hoc multistep comparisons (Tukey). Results: SMH indicated significant (p < 0.01) remineralization only with theobromine (38 ± 32%) and toothpaste (29 ± 16%). With TMR (Δz/lD), theobromine and toothpaste exhibited significantly (p < 0.01) higher mineral gain relative to artificial saliva. With SMH and TMR, remineralization produced by theobromine and toothpaste was not significantly different. With EDS, calcium deposition was significant in all groups, but not significantly different among the groups (theobromine 13 ± 8%, toothpaste 10 ± 5%, and artificial saliva 6 ± 8%). Conclusion: The present study demonstrated that theobromine in an apatite-forming medium can enhance the remineralization potential of the medium.
... Carob is sold in the USA and other Western countries in the health food stores as a substitute for cocoa. It is mainly characterized by high sugar, low fat and high tannin and crude fiber content (Yousif & Alghzawi, 2000) but the complete absence of caffeine and theobromine (Craig & Nguyen, 1984), so in general, it could be considered as a natural sweetener and a natural healthy food. Differences in composition of 10 investigated experimental biscuits are listed in Table 1 ...
Article
The effect of modifying the standard recipe of wheat flour based tea–biscuit on phytate levels, iron content and in vitro availability was investigated. Standard recipe was enriched by addition of dietary fibers and integral raw materials. The average phytic acid content of investigated biscuits ranged from 0.138 to 1.084 g/100 g dry matter of biscuit, depending on biscuit composition. Phytic acid levels were also determined in dough in order to determine the influence of technological process on phytic acid content. Iron content of investigated samples ranged from 0.655 to 4.222 mg/100 g biscuit, and iron availability varied from 26% up to 56%. Data analysis showed that changes in sample composition resulted in significant changes in phytic acid, total and available iron content related to standard sample.
Chapter
Carob (Ceratonia siliqua L.) tree is considered among the most important forest-fruit species native to the Mediterranean region. It has various uses and great valorization potential, all parts of this plant could be exploited as a source of income, as human food or livestock fodder as well as source of raw materials for pharmaceutical, cosmetic, or food industries. Moreover, due to its particular agroecological features, carob tree offers the advantage of growing in poor and unfertile soils in the Mediterranean and Mediterranean-like regions of the world. Thus, carob trees are suitable for the rehabilitation of marginal and sub-marginal areas, helping to compensate for the expanding land desertification in these regions where it can play the role of pioneer and productive species. Carob has been intermittently explored over the last 20 years as a potential tree crop industry in low rainfall areas. The importance of developing the industrial agroforestry potential of the carob tree is hurdled by the lack of options for agroforestry, especially in Mediterranean regions with low rainfall (below 500 mm), and by the need to develop suitable practices for the sustainable management of natural resources. Viable commercial carob cultivation will require mastering efficient farming practices with detailed attention to water requirements and soil fertility. It would improve agricultural productivity in low rainfall areas, help manage water and land degradation, diversify farmers’ incomes, and contribute to the development of export industries contributing to balance the economy of the country. This chapter will provide current knowledge regarding the use of mycorrhizal symbiosis for the improvement of carob culture and productivity in the context of Mediterranean ecosystems. An overview on the multipurpose potential of the carob tree and how spreading its cultivation will benefit people and the environment in marginal areas is highlighted.
Article
The thiolysis of B-type proanthocyanidins in cocoa by cysteamine was evaluated and optimized for its application in cocoa proanthocyanidin quantification. Four thiolysis products consisting of epicatechin, catechin, and their thioethers formed with cysteamine were separated and characterized by reversed-phase UPLC with photo diode array (PDA) detection and high-resolution mass spectrometry (HRMS). A thiolysis time of 20 min under 60 °C temperature was determined as the optimal condition for cocoa proanthocyanidin depolymerization. The optimized thiolysis condition was applied to four cocoa bean samples for proanthocyanidin quantification, using commercially available procyanidin B2 dimer as a reference standard. Satisfactory linearity and quantification and detection limits were achieved for the calibration curves, and proanthocyanidin contents determined by thiolysis were found to be higher than those determined by a published method based on normal-phase HPLC with fluorescence detection. Results in this study suggest promising application potential of cysteamine as an odorless thiolysis agent in routine quantitative analysis of B-type proanthocyanidins. Graphical abstract
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Theobromine, a naturally occurring substance, can be conceived as a prospective inhibitor for uric acid clustering. In aqueous solution, aggregates of π-stacked uric acid molecules with the larger size of clusters are modified into lower-order clusters with a substantial percentage of monomer by the incorporation of theobromine. The composite made of theobromine-uric acid is expected to have enhanced water solubility, allowing stable kidney stones to be excreted through urine. Interestingly, the strategy for the decomposition with feasible modifications in melamine-uric acid composites (that are hydrogen-bonded) is developed (by implementing cluster structure analysis technique and binding free energies). The all-atom molecular dynamics (MD) data provides new insights into the structure and dynamics of uric acid along with melamine molecules in the context of aggregation. The simulation in the present study is supported further by structural and dynamical properties calculations. The calculations of hydrogen bond dynamics, the average number of hydrogen bonds, dimer existence autocorrelation functions, umbrella sampling, and coordination number theorize that the incorporation of theobromine significantly modifies the aggregated structure of uric acid. The overall complexation energy, along with the quantum chemical calculations, further explain the alternation of aggregated structure. Furthermore, the preferential interaction parameter describes at which concentration theobromine-uric acid interaction (which is π-stacked) predominates over uric acid-uric acid interactions. Interestingly, the interactions between theobromine-melamine and melamine-melamine (which are hydrogen-bonded) are not relevant here. Thus, melamine-uric acid cluster size is reduced owing to the disintegration of self-aggregated uric acid clusters by the involvement of theobromine. Moreover, an excellent agreement is observed between present MD results and experimentally obtained data.
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Objective To identify in vitro inhibitors of xanthine crystallization that have potential for inhibiting the formation of xanthine crystals in urine and preventing the development of the renal calculi in patients with xanthinuria. Methods The formation of xanthine crystals in synthetic urine and the effects of 10 potential crystallization inhibitors were assessed using a kinetic turbidimetric system with a photometer. The maximum concentration tested for each compound was: 20 mg/L for 3-methylxanthine (3-MX); 40 mg/L for 7-methylxanthine (7-MX), 1-methylxanthine (1-MX), theobromine (TB), theophylline, paraxanthine, and caffeine; 45 mg/L for 1-methyluric acid; 80 mg/L for 1,3-dimethyluric acid; and 200 mg/L for hypoxanthine. Scanning electron microscopy was used to examine the morphology of the crystals formed when inhibitory effects were observed. Results Only 7-MX, 3-MX, and 1-MX significantly inhibited xanthine crystallization at the tested concentrations. Mixtures of inhibitors had an additive effect rather than a synergistic effect on crystallization. Conclusion Two of the inhibitors identified here—7-MX and 3-MX—are major metabolites of TB. In particular, after TB consumption, 20% is excreted in the urine as TB, 21.5% as 3-MX, and 36% as 7-MX. Thus, consumption of theobromine could protect patients with xanthinuria from the development of renal xanthine calculi. Clinical trials are necessary to demonstrate these effects in vivo.
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Following a request from the European Commission, the EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids (CEF) was requested to deliver a scientific opinion on the safety assessment of the flavouring substances caffeine [FL-no: 16.016] and theobromine [FL-no: 16.032] in the Flavouring Group Evaluation 49, Revision 1. Consequent to the 2015 scientific opinion from the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) on the safety of caffeine from all dietary sources, the CEF Panel considered it inappropriate to evaluate the two substances through the Procedure. For caffeine, the Panel based its assessment on the safety threshold of 5.7 mg/kg body weight (bw) per day for adults, except pregnant/lactating women, and 3 mg/kg bw per day for children, adolescents, pregnant and lactating women, as established by the NDA Panel. The safety evaluation of theobromine takes into account that approximately 11% of an oral dose of caffeine is metabolised to theobromine and that both substances have a similar pharmacological profile. For the exposure assessment, a brand loyalty model was chosen. In this model, it was assumed that a consumer is exposed on a long-term basis to a specific category of food (i.e. non-alcoholic beverages), containing caffeine or theobromine at their respective maximum use levels. For the rest of the categories, normal use levels applied. Daily dietary exposure to caffeine and theobromine (excluding systemic exposure) added as a chemically defined flavouring substance ranged 0–2.3 and 0–0.4 mg/kg bw, respectively, across all population groups. The Panel concluded that caffeine [FL-no: 16.016] and theobromine [FL-no: 16.032] would not be expected to present safety concern based on their estimated levels of intake from their use as flavouring substances.
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Theobromine (THB) is one of the major xanthine-like alkaloids found in cacao plant and a variety of other foodstuffs such as tea leaves, guarana and cola nuts. Historically, THB and its derivatives have been utilized to treat cardiac and circulatory disorders, drug-induced nephrotoxicity, proteinuria and as an immune-modulator. Our previous work demonstrated that THB has the capacity to improve the formation of hydroxyl-apatite during tooth development, suggesting that it may also enhance skeletal development. With its excellent safety profile and resistance to pharmacokinetic elimination, we reasoned that it might be an excellent natural osteoanabolic supplement during pregnancy, lactation and early postnatal growth. To determine whether THB had an effect on human osteoprogenitors, we subjected primary human bone marrow mesenchymal stem cells (hMSCs) to osteogenic assays after exposure to THB in vitro and observed that THB exposure increased the rate of osteogenesis and mineralization by hMSCs. Moreover, THB exposure resulted in a list of upregulated mRNA transcripts that best matched an osteogenic tissue expression signature as compared to other tissue expression signatures archived in several databases. To determine whether oral administration of THB resulted in improved skeletal growth, we provided pregnant rats with chow supplemented with THB during pregnancy and lactation. After weaning, offspring received THB continuously until postnatal day 50 (approximately 10 mg kg⁻¹ day⁻¹). Administration of THB resulted in neonates with larger bones, and 50-day-old offspring accumulated greater body mass, longer and thicker femora and superior tibial trabecular parameters. The accelerated growth did not adversely affect the strength and resilience of the bones. These results indicate that THB increases the osteogenic potential of bone marrow osteoprogenitors, and dietary supplementation of a safe dose of THB to expectant mothers and during the postnatal period could accelerate skeletal development in their offspring.
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Throughout the livestock industry in the western United States, control of canid predators was considered to be of considerable importance to the livestock industry, especially to sheep producers, who sufkred high losses from coyotes and wolves. In the 19th century, the demand for predator control was communicated to Congress and the western state assemblies, with the result that predator control was provided in western states by the Fedcral Bureau of Biological Survey in cooperation with state agencies, and by trappers hired by stockmen. Steel traps and poisons were the principal methods used for predator control during the early years of the program. Historically, predacides have been used in the United States primarily to control wolves, coyotes, and red foxes that prey on livestock. Strychnine was employed in the late 1800s and early 1900s to collect wolf carcasses (Quaife, 1973). Strychnine drop baits were employed for coyote and fox control through the 1960s. Drop baio consisted oi'strychnine rablets put in small pieces of perjshable 6ats then placed around unpoisoned decoy carcasses (Robinson, 1962). bleat baits impregnated with a lethal agent, either thallium sulfate or Compound 1080, were used between 1937 and 1972 (Roh~nson, 1942). Currently, three predacides are ~~vnilabfloer use in controlling coyotes, foxes, wild dogs, and arctic fox. This paper will provide a description of these toxicants and the current status of their use in predator control in the United States.
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The carob tree, Ceratonia siliqua L, is a characteristic constituent of the evergreen, "maquis" and "garigue" vegetation type in low-altitude areas in the Mediterranean Basin. All over these territories, this dioecious and thermophilous tree has been extensively cultivated for its pods. These trees are used for both human consumption and as a sugar-rich animal feed. However, currently the main interest is seed production for gum extraction from the endosperm, which is used as a stabilizer in numerous commercial food products. Most cultivars are female, and require the presence of pollen donors in the orchards. The carob is mainly insect-pollinated (flies, bees, and wasps), but is also wind-pollinated, depending on the habitat. The carob tree shows some outstanding features, like rusticity and drought resistance, and produces under reduced orchard management, and thus is well suited for part-time farming. Carob's main limitation is its frost sensitivity. In addition, modern carob orchards start bearing earlier (fourth year after budding) than traditional carob plots, and they increase yield steadily in response to minimum cultural care and deficit irrigation. This crop has received little attention until now, but currently it is being reemphasized as an alternative in dryland (500mm) or in supplementary drip irrigation areas with a Mediterranean climate for diversification and revitalization of coastal agriculture. This chapter describes carob's economic importance worldwide and their genetic resources and it also reviews various aspects of its taxonomy, botany, origin, ecology, properties, uses, diversity, and breeding. All the life cycle descriptions are referred to the northern hemisphere climatic conditions. In addition, a full account of the crop-producing areas, agronomy, limitations, industry market, and prospects is presented.
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The cigarette ingredients cocoa powder, glycerol, and saccharose were investigated regarding their potential effect on the resulting mainstream smoke, i.e., smoke chemistry (Hoffmann analytes), mammalian cell cytotoxicity (Neutral Red Uptake assay), and bacterial mutagenicity (Ames assay). Each ingredient was added at three concentrations to the tobacco of a 6 mg and 10 mg 'tar' yield experimental American blend filter cigarette (obtained under ISO/FTC smoking regime). The lowest application concentration was equivalent to the normal approximate use level of the ingredients; the highest application level was up to 5-fold higher. The resulting data were compared with the respective control cigarettes without addition of the ingredients. The addition of cocoa powder did not lead to any consistent effects on the measured mainstream smoke analytes. Neither the in vitro cytotoxicity nor the in vitro mutagenicity was affected by cocoa addition. The addition of glycerol resulted in a decrease in the delivery of several smoke constituents (generally around 20%), e.g. aldehydes, phenolics, and N-nitrosamines. Water in the particulate phase (TPM) was distinctly increased (up to +150%). The cytotoxicity of the TPM was decreased (approx.!15%). Mutagenicity was not affected. Saccharose addition consistently increased formaldehyde delivery in smoke by up to 40% and decreased tobacco-specific N-nitrosamines by up to approximately 20%. The increase in formaldehyde is discussed in the context of the human smoker. The cytotoxicity was not affected by the addition of saccharose, while the mutagenicity of the TPM was decreased in tester strain TA98 with metabolic activation (!15%). The results are in agreement with currently available literature. Some investigations summarized in this publication are novel and have not yet been reported in the literature. Based on the total evidence, it can be concluded that the three ingredients added at their current use levels do not increase the inherent toxicity of the cigarette smoke.
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Caffeine is a widely known food constituent. Coffee and tea are major sources of caffeine. However, there is little data about caffeine's antimicrobial activity. Although caffeine is safe for the consumer and suitable as an antimicrobial agent in food, ingesting a lot of caffeine will affect the body (>400 mg day-1). As a result, legislation has defined the amount of caffeine to be used in various food products. In term of antimicrobial activity, caffeine at concentration from 62.5 to >2,000 μg•ml-1 could inhibit bacteria. In addition, higher amount of caffeine (>5,000 μg•ml-1) was found to inhibit mold growth.
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To validate a simple method of urinary theobromine determination, to assess urinary theobromine levels in 80 healthy children and to relate these levels to consumption of cocoa products. Urine samples were diluted, directly injected into an HPLC system, separated by gradient elution on a C18 column, and detected by UV spectrometry. The method was validated for linearity, limits of detection and quantification, imprecision, accuracy, recovery and interferences. The proposed method was used to assess 12-h day and 12-h night urinary theobromine excretion by 80 healthy children, divided into four groups based on consumption of cocoa products. In addition, urinary excretion of magnesium and oxalate, also present in cocoa, were measured in these four groups. The method was linear to a theobromine concentration of 278μmol/L (50mg/L). LOD and LOQ for urine samples, diluted 1:5 (vol/vol) with water, were 1.1 and 3.6μmol/L respectively. Within-run and between-run imprecisions (CV) were each <2%. Average recovery was 99%, and analysis of a certified reference sample showed an error <2.5%. Theobromine excretion levels were significantly higher in healthy children with higher consumption of cocoa products (p<0.001), but oxalate (p=0.098) and magnesium (p=0.068) excretion levels did not differ significantly. This validated method resulted in urinary theobromine determination with 100% recovery, without sample pretreatment. Urinary theobromine levels in healthy children were directly related to their consumption of cocoa products. Copyright © 2015. Published by Elsevier Inc.
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Toxicology is a vast subject. Animals are exposed to numerous drugs, household products, plants, chemicals, pesticides and venomous animals. In addition to the individual toxicity of the various potential poisons, there is also the question of individual response and, more importantly, of species differences in toxicity. This review serves to address some of the common questions asked when dealing with animals with possible poisoning, providing evidence where available. The role of emetics, activated charcoal and lipid infusion in the management of poisoning in animals, the toxic dose of chocolate, grapes and dried fruit in dogs, the use of antidotes in paracetamol poisoning, timing of antidotal therapy in ethylene glycol toxicosis and whether lilies are toxic to dogs are discussed. © 2015 British Small Animal Veterinary Association.
Article
Several concentrations of theobromine (TB) and (-)-epicatechin (EC) were coadministered to rats, and plasma EC and its metabolites were determined using ultra-high-performance liquid chromatography-tandem mass spectrometry. It has been demonstrated that TB increases the absorption of EC in a dose-dependent manner. Cocoa powder had a similar effect, and the mechanism involved is not thought to depend on tight junctions.
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Method for separation and quantification of methylxanthins, catechins, and theaflavins in Korean commercial teas (green, oolong, and black teas) was developed using reversed phase high-performance liquid chromatography (HPLC). After extraction with hot water, tea compounds were separated on Inertsil ODS-3v column, eluted with gradient of 7% acetonitrile and 93% of 20 mM phosphate buffer mixture for 7 min. Column effluent was monitored at 270 nm. This technique was effective for analyses of m methylxanthins, catechins, and theaflavins in teas and biological samples. In green and oolong teas, two kinds of methylxanthins and 7 of catechins were identified, whereas 4 theaflavins were only identified in black tea. Among seven catechins in green and oolong teas, EGCG showed highest amount, whereas ECG was highest in black tea. (theaflavins were found only in black teas) In all teas, theobromine content was lower than that of caffeine.
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Antioxidant activity, gastrointestinal solubility of polyphenols and formation of different classes of Maillard reaction products (MRPs) that occurs during thermal processing of carob powder was investigated in relation to different roasting time-roasting temperature combinations applied. Principal component analysis (PCA) was used to group carob powders with the aim of proposing optimal roasting conditions for obtaining the one with the highest biological activity that can be expected after undergoing the simulation of gastrointestinal digestion. Results showed that antioxidant activity of carob powder can be significantly increased by roasting due to improved polyphenol solubility and formation of the early-stage MRPs. Fluorescent MRPs, HMF and furfural, known for their toxic properties, were the most abundant in samples roasted under more severe time/temperature conditions. Carob powder roasted at 130 °C for 30 min yielded the highest antioxidant activity and the lowest toxic MRP formation.
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Conservation biology is called a "crisis discipline." In a world undergoing rapid change, this science informs us about research, technologies, management practices, and policies that can help protect the earth's naturally-occurring biological diversity. The six chapters of this book provide insightful analysis on managing protected areas (Middle East), conserving biochemical and genetic diversity of carob tree (Tunisia) and wild pear (Japan), determining the health status of Amazon manatee, manipulating sex ratios to benefit wildlife, and narrowing the gap between religion and conservation. The authors approach threats to biological diversity from varied angles, reflecting the interdisciplinary nature of the field. This book offers room for reflection on the definition and utility of the word 'natural' on a planet now overwhelmingly dominated by people.
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The interactions between some xanthine derivatives: caffeine (Cf), theophylline (Tp), theobromine (Tb), and histamine (Hm) with phytate (Phy) were studied at T = 298.15 K and I = 0.5 mol L− 1 in aqueous NaCl solution using ISE-[H+] potentiometry. For all of the investigated systems, [(Phy)(L)Hq](q − 7) species (q = 4 to 9, depending on the maximum protonation degree of the ligand) were formed in high yields (30–90%). The stability constants of the systems varied; higher stability was observed for the [(Phy)(Tb)Hq](q − 7) (5 ≤ q ≤ 8) species. For the [(Phy)(Tb)H8]+ and [(Phy)(Tb)H7] species, we obtained values of log K8 = 8.1 and log K7 = 5.58, respectively. The formation ΔH and TΔS parameters were determined for the Phy–Cf system at I = 0.5 mol L− 1 in NaCl(aq) by calorimetric titrations. As an example, for the [(Phy)(Cf)H5]2 − species, we obtained the following values: ΔH = − 4.0 ± 0.2 kJ mol− 1, − ΔG = 14.6 ± 0.5 kJ mol− 1 and TΔS = 10.5 ± 0.8 kJ mol− 1.
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A knowledge of the physical characteristics of the carob pod and kernels is of great importance in their industrial use. This work provides contributions to knowledge of the Spanish carob tree population, based on the morphological characteristics of the carob pod and the yield of the kernels. To this end, Spanish regions in which the carob tree is cultivated were sampled. The total number of carob trees sampled was 182, located in the Balearic Islands and near the Mediterranean Coast. The results indicate a great similarity between Spanish carob pods and those of other Mediterranean countries, especially Italy and Cyprus. Correlating the different measurements obtained indicates that to achieve high yields of kernels and of carob gum, pods which are narrow and thin and/or kernels which are heavy, fat and short need to be harvested.
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This is one of the more than 20 crop monographs that a BMZ, Germany funded and IPGRI coordinated project has produced on the promotion of the conservation and use of neglected and underutilized crops. For more details see the full text.
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An investigation is hereby presented on the possible formation of a stacking (π–π) complex between adenosine-5′-triphosphate (ATP) and three xanthines (L), namely caffeine (Cf), theophylline (Tp) and theobromine (Tb), at different ionic strengths (0.05 ≤ I/mol L−1 ≤ 1) in NaCl(aq) and temperature (T = 283.15, 298.15 and 313.15 K), by using magnetic resonance spectroscopy 1H NMR. The values of log β of the [ATP(L)]4− species were: 1.32, 1.24 and ≤0.7 at I = 0.05 mol L−1 and T = 298.15 K for Cf, Tp and Tb, respectively. The stability constant of [ATP(Cf)]4− species shown a linear dependence on the temperature, and the values of log β obtained were: 1.18, 1.32 and 1.47 at T = 283.15, 298.15 and 313.15 K, respectively. By using the values of log β obtained at different temperatures a value of ΔH = −16 ± 3 kJ mol−1 was calculated for the formation of the stacking (π–π) complex species. The three systems were also studied potentiometrically at different ionic strengths (0.05 ≤ I/mol L−1 ≤ 1) in NaCl(aq) and T = 298.15 K. For all the investigated systems, the formation of [(ATP)(L)Hi](i−4) species (i = 1–3), with high yields (60–80%) was found; the stability trend obtained for the protonated complex species was (ATP–Tb) > (ATP–Tp) > (ATP–Cf). The stability constants of all the [(ATP)(L)Hi](i−4) species proved fairly dependent on the ionic strength that was studied by using the Debye–Hückel-type equation. Furthermore, by comparison with the stability of the species of analogous systems, such as ATP–aliphatic amines, ATP–aromatic amines and pyrophosphate–aliphatic amines, an empirical relationship between the stability of the complexes and the total basicity of the ligand was found.Graphical abstractHighlights► The interaction of ATP and xanthines was studied by 1H NMR and potentiometry. ► π–π stacking was evidenced between ATP and xanthines. ► The interactions were characterized quantitatively. ► The dependence on ionic strength and on temperature was determined.
Article
Near-infrared (NIR) reflectance spectroscopy was used to develop calibration equations to measure chemical composition (dry matter, crude protein and reducing sugars) in carob samples. Calibration equations were obtained by multiple linear regression of conventional laboratory analysis values on NIR spectra from 75 samples and verified with 20 additional samples from the original population, collected on the Spanish Mediterranean littoral in different years. A Tecnichon Infra Alyzer model 450 monochromator interfaced with an IBM PS2 microcomputer was used for the study. Two alternative calibrations were chosen using three and six wavelengths. The coefficients of multiple correlation ranged from 0·91 to 0·95 and the standard errors of calibration ranged from 0·38 to 1·62. The study showed that NIR analysis can be used for predicting the chemical composition of carob pulp.
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European badgers (Meles meles) are a wildlife reservoir for Mycobacterium bovis infection (tuberculosis) in Ireland and the UK and are implicated in the transmission of infection to livestock. Vaccination of badgers with the human BCG vaccine (Bacille Calmette Guerin) is considered as an important strategy to reduce the burden of disease in this species, and a pragmatic approach is likely to involve oral vaccination. In this study, we evaluated nine different flavours for use as attractants in a prototype oral vaccine bait for European badgers (M. meles): aniseed, apple, cocoa powder, carob powder, curry, fish, garlic, peanut and strawberry. The bait matrix was composed of a natural lipid formulation, developed as a vehicle for oral vaccination against tuberculosis in wildlife. A ‘food for work’ paradigm was employed during the trials to ensure the animals were actively seeking the baits. The trials showed carob and cocoa powders were equally attractive and more attractive than any of the other candidates. Carob and cocoa show potential as bait attractants for badgers and might form part of a novel vaccine delivery system. KeywordsAttractants–Carob–European badger– Meles meles –Oral vaccine–Bovine tuberculosis
Article
Ginseng is not widely accepted by U.S. consumers due to its unfamiliar flavors, despite its numerous health benefits. Previous studies have suggested that the bitter compounds in chocolate and coffee may mask the off-flavors of ginseng. The objectives of this study were to: (1) profile sensory characteristics of ginseng extract solution, caffeine solution, cyclo (L-Pro-L-Val) solution, theobromine solution, and 2 model solutions simulating chocolate bitterness; and (2) determine the changes in the sensory characteristics of ginseng extract solution by the addition of the bitter compounds found in chocolate and coffee. Thirteen solutions were prepared in concentrations similar to the levels of the bitter compounds found in coffee and chocolate products. Twelve panelists participated in a descriptive analysis panel which included time-intensity ratings. Ginseng extract was characterized as sweeter, starchier, and more green tea than the other sample solutions. Those characteristics of ginseng extract were effectively modified by the addition of caffeine, cyclo (L-Pro-L-Val), and 2 model solutions. A model solution simulating dark chocolate bitterness was the least influenced in intensities of bitterness by the addition of ginseng extract. Results from time-intensity ratings show that the addition of ginseng extract increased duration time in certain bitterness of the 2 model solutions. Bitter compounds found in dark chocolate could be proposed to effectively mask the unique flavors of ginseng. Future studies blending aroma compounds of chocolate and coffee into such model solutions may be conducted to investigate the influence on the perception of the unique flavors through the congruent flavors.
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The effects of theobromine in man are underresearched, possibly owing to the assumption that it is behaviourally inert. Toxicology research in animals may appear to provide alarming results, but these cannot be extrapolated to humans for a number of reasons. Domestic animals and animals used for racing competitions need to be guarded from chocolate and cocoa-containing foods, including foods containing cocoa husks. Research ought to include caffeine as a comparative agent, and underlying mechanisms need to be further explored. Of all constituents proposed to play a role in our liking for chocolate, caffeine is the most convincing, though a role for theobromine cannot be ruled out. Most other substances are unlikely to exude a psychopharmacological effect owing to extremely low concentrations or the inability to reach the blood-brain barrier, whilst chocolate craving and addiction need to be explained by means of a culturally determined ambivalence towards chocolate.
Article
Large-scale production has been the major obstacle to the success of many biopesticides. The spreading of microbial biocontrol agents against postharvest disease, as a safe and environmentally friendly alternative to synthetic fungicides, is quite dependent on their industrial mass production from low-cost raw materials. Considerable interest has been shown in using agricultural waste products and by-products from food industry as nitrogen and carbon sources. In this work, carob pulp aqueous extracts were used as carbon source in the production of the biocontrol agent Pantoea agglomerans PBC-1. Optimal sugar extraction was achieved at a solid/liquid ratio of 1:10 (w/v), at 25°C, for 1 h. Batch experiments were performed in shake flasks, at different concentrations and in stirred reactors at two initial inoculums concentrations, 10(6) and 10(7) cfu ml(-1). The initial sugar concentration of 5 g l(-1) allowed rapid growth (0.16 h(-1)) and high biomass productivity (0.28 g l(-1) h(-1)) and was chosen as the value for use in stirred reactor experiments. After 22 and 32 h of fermentation the viable population reached was 3.2 × 10(9) and 6.2 × 10(9) cfu ml(-1) in the fermenter inoculated at 10(6) cfu ml(-1) and 2.7 × 10(9) and 6.7 × 10(9) cfu ml(-1) in the bioreactor inoculated at 10(7) cfu ml(-1). A 78% reduction of the pathogen incidence was achieved with PBC-1 at 1 × 10(8) cfu ml(-1), grown in medium with carob extracts, on artificially wounded apples stored after 7 days at 25°C against P. expansum.
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An improved chromatographic method was developed using ultra-performance liquid chromatography-tandem mass spectrometry to identify and quantify phenolic compounds and alkaloids, theobromine and caffeine, in carob flour samples. The developed method has been validated in terms of speed, sensitivity, selectivity, peak efficiency, linearity, reproducibility, limits of detection, and limits of quantification. The chromatographic method allows the identification and quantification of 20 phenolic compounds, that is, phenolic acids, flavonoids, and their aglycone and glucoside forms, together with the determination of the alkaloids, caffeine and theobromine, at low concentration levels all in a short analysis time of less than 20 min.
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Chocolate contains a wide range of antioxidants that includes soluble phenolic compounds (phenolic acids, catechin, epicatechin, and proanthocyanidins), insoluble polymeric phenolics and methylxanthines. The objective of this study was to determine phenolic and theobromine contents in dark (DC), milk (MC), and white (WC) chocolates commonly found in the Malaysian marketplace. Total phenolic and flavonoids were determined by means of a spectrometric assay, while catechin, epicatechin and theobromine were quantified using a reverse-phase HPLC method. Dark chocolates exhibited the highest phenolics and flavonoids contents, followed by milk and white chocolates. Catechin and epicatechin were major flavonoids detected in dark chocolates. Theobromine was detected in dark and milk chocolates, but not in white chocolates. A high correlation (r= 0.93) between total phenolic and flavonoid contents, indicating that the major phenolic compounds in dark chocolates belong to the flavonoid class. When nutrition and health promotion are of concern, dark chocolates would be recommended over milk and white chocolates owing to their higher contents of antioxidant phenolic compounds.
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A reverse-phase liquid chromatography analysis is used to access the quantity of theobromine, (+)-catechin, caffeine, and (-)-epicatechin in Standard Reference Material 2384 Baking Chocolate, cocoa, cocoa beans, and cocoa butter using water or a portion of the mobile phase as the extract. The procedure requires minimal sample preparation. Theobromine, (+)-catechin, caffeine, and (-)-epicatechin are detected by UV absorption at 273 nm after separation using a 0.3% acetic acid-methanol gradient (volume fractions) and quantified using external standards. The limit of detection for theobromine, (+)-catechin, caffeine, and (-)-epicatechin averages 0.08, 0.06, 0.06, and 0.06 microg/mL, respectively. The method when applied to Standard Reference Material 2384 Baking Chocolate; baking chocolate reference material yields results that compare to two different, separate procedures. Theobromine ranges from 26000 mg/kg in cocoa to 140 mg/kg in cocoa butter; (+)-catechin from 1800 mg/kg in cocoa to below detection limits of < 32 mg/kg in cocoa butter; caffeine from 2400 mg/kg in cocoa to 400 mg/kg in cocoa butter, and (-)-epicatechin from 3200 mg/kg in cocoa to BDL, < 27 mg/kg, in cocoa butter. The mean recoveries from cocoa are 102.4 +/- 0.6% for theobromine, 100.0 +/- 0.6 for (+)-catechin, 96.2 +/- 2.1 for caffeine, and 106.2 +/- 1.7 for (-)-epicatechin.
Article
Theobromine (TB) is a significant pharmacological ingredient for chocolate (C). 10 nursing mothers (ages 25 to 32) who were nursing 1-24 mos were studied. Other methylxanthines were avoided. Each mother ate 1 C bar (1.2 ozs) containing 60 mg TB and 5 mg caffeine (Caf). Simultaneous saliva and milk samples were collected at 0, 1, 2, 3, 5, 8, 12 and 15 hrs. Urine was collected on the infants for 3-6 hrs after maternal dosing. Samples were assayed for TB and Caf using HPLC with mobile phase 0.01 M Na acetate pH 4.0-acetronitrile (93:10) using theophylline as the internal standard. TB appeared in both milk and saliva by 1 hr and peaked in all but 1 pt by 3 hrs. Paired samples showed higher amounts of TB in milk in all but two pts. Saliva/milk ratios ranged from 0.7-1.3 during first 4 hours. Elimination phase t½ ranged from 4.9-31.5 (x = 14.7±8.16) hrs for milk and 3.8-33.0 (x = 9.80±8.65) hrs for saliva. There was not agreement between milk and saliva values for 6/10 patients. Assuming each infant would ingest 90 ml milk every 3 hrs for 24 hrs after maternal ingestion of C, the amount of TB available for ingestion ranged from 0.44−1.68 (x = 1.06±0.47) mg or 0.73−2.80 (x = 1.77±0.79) % of maternal dose. In urine of nursing infants, levels of TB were below 0.6 μg/ml; no Caf was detected. There were no untoward symptoms in the infants. Usual intakes of chocolate does not appear to present significant doses of either TB nor Caf to the nursing infant.
Article
Eighteen randomly selected cocoa samples were analyzed in duplicate by HPLC for caffeine and theobromine. The relative standard deviation for the method was determined to be 1.55%. The mean ± S.D. of the theobromine plus caffeine content (moisture free basis) was 2.85 ± 0.08% with a 90% confidence interval of 2.72–2.98% for red Dutched cocoa. The total ± SD. for natural cocoas was 2.82 + 0.16% with a 90% confidence interval of 2.56–3.08%.
Article
Commercial chocolate products were analyzed for theobromine and caffeine content by High Performance Liquid Chromatography, Levels in 22 samples of chocolate liquor averaged 1.22% theobromine and 0.21% caffeine. Commercial cocoas contained, on the average, 1.89% theobromine and 0.21% caffeine. Sweet chocolate averaged 0.46% theobromine and 0.07% caffeine, while milk chocolate averaged 0.15% theobromine and 0.02% caffeine. Hot cocoa (chocolate) beverages averaged 65 mg of theobromine and 4 mg of caffeine per 5 ounce serving and chocolate milk prepared from a variety of cocoa-sugar mixes averaged 58 mg of theobromine and 5 mg of caffeine per 8 ounce serving. Theobromine and caffeine levels varied widely in individual samples within the product categories examined. Also, the ratio of theobromine to caffeine varied widely among different chocolate liquors ranging from a low of 2.5:1 to a high of 23.0:1.
Article
Popular brands of instant and ground coffee, instant and bag tea, and instant hot cocoa mixes were prepared according to manufacturer's directions and analyzed for caffeine and/or theobromine content. After extraction into boiling water, the methylxanthines were separated by high performance liquid chromatography using a reverse phase C18 column and a mobile phase of acetonitrile and water (8:92, v/v). Caffeine content ranged from 32.4–35.0 mg/cup in instant tea, 30.2–67.4 mg/cup in bag tea, 1.0–7.8 mg/cup in instant hot cocoa mixes, 46.7–67.6 mg/cup in instant coffee, and 93.0–163.5 mg/cup in ground coffee. Theobromine concentrations ranged from 1.4–2.3 mg/cup in instant tea, 1.2–4.4 mg/cup in bag tea, and 39.5–79.5 mg/cup in instant hot cocoa mixes.
A method was developed for determining theobromine and caeffine in cocoa and chocolate products by high pressure liquid chromatography. After a simple hot water extraction, both theobromine and caffeine were separated by using a reverse phase C18 column and a mobile phase of methanol-water-acetic acid (20 + 79 + 1). Theobromine and caffeine were quantitated at 280 nm; average recoveries were 98.7 and 95.0%; and coefficients of variation were 2.31 and 3.91%, respectively.
Article
Six nursing mothers ingested 113 gm of Hershey's milk chocolate containing 240 mg of theobromine. Samples of plasma, saliva, and breast milk were assayed for theobromine by high pressure liquid chromatography. Peak theobromine concentrations of 3.7 to 8.2 mg/l were found in all fluids at 2 to 3 hour after ingestion of chocolate. The disposition half-life of theobromine averaged 7.1 +/- 2.1 hours, body clearance was 65 +/- 20 ml/hour/kilogram, and the apparent volume of distribution was 0.62 +/- 0.13 l/lk. Theobromine is only slightly bound to plasma and milk proteins and concentrations in milk and saliva matched plasma data closely. The mean concentration ratios were 0.82 +/- 0.17 for milk/plasma and 0.92 +/- 0.17 for saliva/plasma. If a mother ate a 4-ounce chocolate bar every 6 hours and the infant nursed when the theobromine concentration in milk was at its peak, the infant could ingest about 10 mg of theobromine per day.
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The developing fetus is often subjected to the influence of various drugs ingested by the mother during gestation. Currently, caffeine is one of the more common drugs used by the pregnant patient. It is important that the patient be made aware that caffeine is a drug and that it may cause potential harm to the fetus. Until more scientific data are available, it is our recommendation that moderate to excessive use of caffeine during pregnancy be curtailed and that mild use be decided on an individual basis.
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The use of methylxanthine molecules labeled with stable isotopes (15CN and 13C) gives the following results: 1. The biotransformation of theophylline to caffeine by N-7 methylation occurs in premature newborns treated with this methylxanthine. 2. It permits us to differentiate with perfect selectivity the two origins of caffeine: exogenous caffeine coming from breast feeding and endogenous caffeine coming from the infant's metabolism. 3. It allows us to measure the placental transfer of theophylline administered to the mother and to show that the fetus can metabolize this xanthine into caffeine.
Four duplicate samples of cocoa-containing materials, a practice sample, and standards were submitted to the collaborators for theobromine and caffeine analysis by HPLC. In the method the samples are defatted with petroleum ether, and dried. The fat-free residue is then extracted with water and an aliquot is injected into the chromatograph. Compounds are quantitated by comparison with internal or external standards, either by peak height or peak area. Results for all the analyses showed that few of the values were more than 2 standard deviations from the mean. The method has been adopted as official first action.