Showing papers on "Theobromine published in 2009"

Two Distinct Pathways for Metabolism of Theophylline and Caffeine Are Coexpressed in Pseudomonas putida CBB5

Abstract: Pseudomonas putida CBB5 was isolated from soil by enrichment on caffeine. This strain used not only caffeine, theobromine, paraxanthine, and 7-methylxanthine as sole carbon and nitrogen sources but also theophylline and 3-methylxanthine. Analyses of metabolites in spent media and resting cell suspensions confirmed that CBB5 initially N demethylated theophylline via a hitherto unreported pathway to 1- and 3-methylxanthines. NAD(P)H-dependent conversion of theophylline to 1- and 3-methylxanthines was also detected in the crude cell extracts of theophylline-grown CBB5. 1-Methylxanthine and 3-methylxanthine were subsequently N demethylated to xanthine. CBB5 also oxidized theophylline and 1- and 3-methylxanthines to 1,3-dimethyluric acid and 1- and 3-methyluric acids, respectively. However, these methyluric acids were not metabolized further. A broad-substrate-range xanthine-oxidizing enzyme was responsible for the formation of these methyluric acids. In contrast, CBB5 metabolized caffeine to theobromine (major metabolite) and paraxanthine (minor metabolite). These dimethylxanthines were further N demethylated to xanthine via 7-methylxanthine. Theobromine-, paraxanthine-, and 7-methylxanthine-grown cells also metabolized all of the methylxanthines mentioned above via the same pathway. Thus, the theophylline and caffeine N-demethylation pathways converged at xanthine via different methylxanthine intermediates. Xanthine was eventually oxidized to uric acid. Enzymes involved in theophylline and caffeine degradation were coexpressed when CBB5 was grown on theophylline or on caffeine or its metabolites. However, 3-methylxanthine-grown CBB5 cells did not metabolize caffeine, whereas theophylline was metabolized at much reduced levels to only methyluric acids. To our knowledge, this is the first report of theophylline N demethylation and coexpression of distinct pathways for caffeine and theophylline degradation in bacteria.

Rapid determination of phenolic compounds and alkaloids of carob flour by improved liquid chromatography tandem mass spectrometry.

Abstract: 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.

Identification of paraxanthine as the most potent caffeine-derived inhibitor of connective tissue growth factor expression in liver parenchymal cells.

Abstract: Background: Recently, we identified hepatocytes as the major cellular source of profibrogenic connective tissue growth factor (CTGF/CCN2) in the liver. Based on reports of a hepatoprotective effect of coffee consumption, we were the first to provide evidence that caffeine suppresses transforming growth factor (TGF)-β dependent and -independent CTGF expression in hepatocytes in vitro and in vivo, thus suggesting this xanthine-alkaloid as a potential therapeutic agent. Aim: This study aims at comparing the inhibitory capacities of caffeine and its three demethylated derivates paraxanthine, theophylline and theobromine on CTGF expression in hepatocytes and hepatic stellate cells (HSC). Results: Our data suggest paraxanthine as the most important pharmacological repressor of hepatocellular CTGF expression among the caffeine-derived metabolic methylxanthines with an inhibitory dosage (ID)50 of 1.15 mM, i.e. 3.84-fold lower than what is observed for caffeine. In addition, paraxanthine displayed the least cell toxicity as proven by the water-soluble tetrazolium-1 cell vitality assay. However, caffeine or any of the metabolites did not inhibit CTGF expression in HSC. At the toxicological threshold concentration of 1 mM for paraxanthine, we observed an inhibition of hepatocellular CTGF synthesis by 44%, which was strongly reverted in the presence of the specific competitive cyclic adenosine monophosphate inhibitor Rp-adenosine 3′,5-cyclic monophosphorothioate triethylammonium salt. Furthermore, CTGF protein expression induced by various concentrations of TGF-β (0.13–1 ng/ml) is still reduced by, on average, 27%/45% in the presence of paraxanthine (1.25 mM/2.5 mM). Conclusion: Our data provide an evidence-based suggestion of the caffeine-derived primary metabolite paraxanthine as a potentially powerful antifibrotic drug by its inhibitory effect on (hepatocellular) CTGF synthesis.

Altered expression of the caffeine synthase gene in a naturally caffeine-free mutant of Coffea arabica

Abstract: In this work, we studied the biosynthesis of caffeine by examining the expression of genes involved in this biosynthetic pathway in coffee fruits containing normal or low levels of this substance. The amplification of gene-specific transcripts during fruit development revealed that low-caffeine fruits had a lower expression of the theobromine synthase and caffeine synthase genes and also contained an extra transcript of the caffeine synthase gene. This extra transcript contained only part of exon 1 and all of exon 3. The sequence of the mutant caffeine synthase gene revealed the substitution of isoleucine for valine in the enzyme active site that probably interfered with enzymatic activity. These findings indicate that the absence of caffeine in these mutants probably resulted from a combination of transcriptional regulation and the presence of mutations in the caffeine synthase amino acid sequence.

Influence of xanthine derivatives on cough and airway reactivity in guinea pigs

Abstract: Xanthine derivatives may inhibit phosphodiesterases without selective action on their single isoforms. In this study, effects of theophylline and theobromine on cough and airway reactivity were evaluated in awake guinea pigs using double-chamber whole body plethysmograph. Pre-treatment with theophylline and theobromine (10 mg/kg, i.p.) decreased the number of cough efforts evoked by inhalation of citric acid aerosol (0.6 mol/l) in both healthy and ovalbumin-sensitized animals. Theophylline and theobromine decreased in vivo airway reactivity, i.e., specific airway resistance measured after nebulization of citric acid and histamine aerosol (10(-6) mol/l), only in ovalbumin-sensitized animals, whereas in vitro reactivity to cumulative doses of histamine and acetylcholine (10(8)-10(-3) mol/l) measured in organ chambers significantly decreased in both healthy and ovalbumin-sensitized animals, with more pronounced effect in the latter group. In conclusion, administration of theophylline and theobromine influenced the cough and airway reactivity in ovalbumin-sensitized guinea pigs, indicating the anti-inflammatory potential of xanthine derivatives.

Occurrence of theobromine synthase genes in purine alkaloid-free species of Camellia plants

Abstract: Caffeine (1,3,7-trimethylxanthine) and theobromine (3,7-dimethylxanthine) are purine alkaloids that are present in high concentrations in plants of some species of Camellia. However, most members of the genus Camellia contain no purine alkaloids. Tracer experiments using [8-(14)C]adenine and [8-(14)C]theobromine showed that the purine alkaloid pathway is not fully functional in leaves of purine alkaloid-free species. In five species of purine alkaloid-free Camellia plants, sufficient evidence was obtained to show the occurrence of genes that are homologous to caffeine synthase. Recombinant enzymes derived from purine alkaloid-free species showed only theobromine synthase activity. Unlike the caffeine synthase gene, these genes were expressed more strongly in mature tissue than in young tissue.

Simultaneous and specific determination of proxyphylline, theophylline and other xanthine derivatives in serum by high pressure liquid chromatography.

Abstract: A HPLC-method for rapid, simultaneous and specific determination of proxyphylline, theophylline, glyphylline, theobromine, caffeine and 3-methylxanthine using 8-chlorotheophylline as internal standard is presented. Using 20 microliter deproteinised serum samples for injection, the the detection limits for theophylline and proxyphylline are about 0.040 and 0.055 microgram per ml serum or plasma, respectively, and the corresponding sensitivities are about 0.033 and 0.048 microgram compound for 0.0044 absorbance units at 280 nm. In duplicate analyses the coefficient of variation is less than 3%. A number of frequently used drugs did not interfere in the analyses of serum samples from patients under treatment or in experiments with drug additions to serum samples.

Detection and characterization of caffeine-less tea plants originated from interspecific hybridization

Abstract: Two tea plants containing little caffeine (caffeine-less) were found in a population derived from natural crossings of an interspecific hybrid ‘Cha Chuukanbohon Nou 6’ (Camellia taliensis × C. sinensis). Two caffeine-less plants showed low caffeine content and rather high theobromine, which is a precursor of caffeine, whereas all tested tea (C. sinensis) cultivars including ‘Cha Chuukanbohon Nou 6’ were observed high caffeine/low theobromine contents. Component analysis by HPLC showed that ‘Taliensis-akame’ (C. taliensis), which is the seed parent of ‘Cha Chuukanbohon Nou 6’, showed low caffeine content and rather high theobromine and may be a caffeine-less character donor. Pedigree analysis using SSR markers suggested that parent-offspring relationship was found between tea plants related to caffeine-less tea breeding lines. Therefore, it is considered that caffeine-less character of caffeine-less plants could be inherited recessively from ‘Taliensis-akame’ via ‘Cha Chuukanbohon Nou 6’. Preliminary genetic analysis using 33 progenies of ‘Cha Chuukanbohon Nou 6’ suggested the possibility that the caffeine-less character might be controlled by one recessive locus. Caffeine-less tea plants found in this study, will be efficiently used for genetic resources to introgress caffeine-less traits to tea cultivars in breeding programs.

Genetic variation of phytochemical compounds in progenies of Ilex paraguariensis St. Hil.

Abstract: Mate (Ilex paraguariensis St. Hil) contains phytochemical compounds capable of preventing a number of health problems. Knowledge on the genetic contribution to the variability in these compounds can help to obtain mate progenies with higher levels thereof in breeding programs. The composition of triterpene saponins, methylxanthines, chlorogenic acid and the antioxidant activity of eight mate progenies were evaluated. Significant differences among progenies were verified in contents of triterpene saponins (0.003-0.080%), caffeine (0.226-1.377%), theobromine (0.176-0.831%), and chlorogenic acid (1.344-2.031%) and in antioxidant activity (31.251-51.406%). The contents of theobromine were found to be negatively correlated with saponins and caffeine, and caffeine with chlorogenic acid, while theobromine was positively correlated with chlorogenic acid. The heritability values for saponins (75.09%), caffeine (75.19%), theobromine (66.87%), chlorogenic acid (52.86%) and antioxidant activity (67.75%) indicate the possibility of genetic gain in selection for these traits.

Bitter alkaloid containing consumables comprising bitter blockers

Abstract: Disclosed are bitter alkaloid-containing consumables comprising bitter blockers for a reduced alkaloid-derived bitterness and methods of forming said consumables. The preferred bitter blockers are trilobatin and hesperetin dihydrochalcone 4 -beta-D-glucoside (HDG). Examples of bitter alkaloids are coffeine, theophylline and theobromine.

Effect of cocoa tea (Camellia ptilophylla) co-administrated with green tea on ambulatory behaviors.

Abstract: We investigated the effects of cocoa tea and its main active compound theobromine on ambulatory activity, compared with green tea and caffeine. Although cocoa tea and theobromine themselves didn't change the ambulatory behaviors as green tea and caffeine did, combined administration with green tea or caffeine showed a synergistic action. The obtained data are perhaps contribution to the consumption of cocoa tea in the world.

[Rapid determination of theophylline, theobromine and caffeine in dietary supplements containing guarana by ultra-performance liquid chromatography].

Abstract: A rapid and simple method for determination of theophylline, theobromine and caffeine in dietary supplements containing guarana by ultra-performance liquid chromatography (UPLC) has been developed. Theophylline, theobromine and caffeine were extracted from finely powdered samples with water in a boiling water bath for 20 min, then the extracts were filtered and the filtrates were subjected to UPLC. Liquid samples were diluted with water and filteres, and the filtrates were subjected to UPLC. UPLC separation was performed on an AQUITY UPLC BEH C18 column (2.1 mm i.d.x50 mm, 1.7 microm, Waters) with 10 mmol/L ammonium acetate buffer (pH 4.0)-acetonitrile gradient and eluates were monitored at 275 nm. The recoveries of theophylline (spiked at 200 microg/g [tablet] and 50 microg/mL [liquid]), theobromine (spiked at 200 microg/g [tablet] and 50 microg/mL [liquid]) and caffeine (spiked at 1,000 microg/g [tablet] and 250 microg/mL [liquid]) were 97.6-98.7%, 97.3-99.7%, 97.1-105.4%, respectively. The quantitation limits of theophylline, theobromine and caffeine were 10 microg/g (seed, seed powder, tablet and capsule) and 2.0 microg/mL (liquid) each. When this analytical method was applied to commercial dietary supplements, theophylline, theobromine and caffeine were found at concentrations of 4.45 mg/tablet, 5.48 mg/tablet, 139 mg/tablet, respectively. Taking 4 tablets of this product according to the directions on the package could be dangerous to consumers because of possible overdosing of these ingredients.

The Effect of Caffeine on the Bacterial Populations in a Freshwater Aquarium System

Abstract: Caffeine is becoming a common chemical found in the environment but, little research has been done to understand the environmental effects of caffeine, including dissolved caffeine in aquatic systems. The goal of this research study was to begin to understand how caffeine may interact with aquatic environment, using the bacteria of the genus Pseudomonas, commonly found in the aquatic habitat. We found that Pseudomonas shows an increase in growth when exposed to caffeine, which results in a change in spectrophotometric absorbance, increase in bacterial colony count in the presence of caffeine, and in bio-film like sheen appearing on the glass of the experimental aquarium. Along with the increased bacterial growth, a dramatic increase in ammonia concentrations was observed. Ammonia is toxic to fish, and can be correlated with the metabolic activity of the Pseudomonas bacteria, making the caffeinated environment toxic for aquatic life. Caffeine is a common chemical found in the environment. An evaluation of 139 stream sites in the U.S. for the occurrence of organic wastewater contaminants (OWCs) found that caffeine was the fourth most frequently detected chemical, and occurred in 70% of the samples (Kolpin et al.,2002). Caffeine is one of the most commonly consumed chemicals in the United States and Europe, with one in three Americans consuming approximately 200mg of caffeine each day (Leonard et al., 1987). Caffeine is an alkaloid found in many plant species. It belongs to the group of compounds known as methylxanthines which include theobromine (cocoa) and theophylline (tea). Because of its excellent solubility and slow rate of degradation, caffeine can persist in aquatic environments (Seiler et al., 2005) and has the potential to biomagnify through the food chain. Thus, even a small amount of dissolved caffeine in aquatic environment can concentrate over time. The fact that caffeine is one of the most common compounds found in sewage has been known for over 20 years (Garrison et al., 1976), but the nature of its effect on the aquatic environment still remains unclear. These effects need to be studied so the appropriate agencies in charge of environmental protection can take steps to manage caffeine presence in the

Frozen Confectionery Products

Abstract: A frozen confection product is provided, the product comprising: from 20 to 80% of a frozen composition comprising cocoa solids; from 20 to 60% of a chocolate or chocolate analogue comprising cocoa solids; and from 5 to 20% (by weight of the product) of cocoa nibs; wherein the product contains at least 400 mg of theobromine and at least 40 mg of caffeine per 100 grams.

Production method and production apparatus for a high theobromine-containing composition

Abstract: An object is to provide a method for producing a high theobromine-containing composition effectively in a simple way. The method for producing a theobromine-containing composition comprises: (a) performing an extraction of a theobromine-containing plant or a processed product thereof with a solvent, to obtain a crude theobromine extract; (b) applying the crude theobromine extract to a cation exchange resin subjected in advance to hydrogen ion substitution, to allow an adsorption of theobromine onto the cation exchange resin; and (c) passing a solvent containing no ionic substance through the cation exchange resin subsequently to the step (b), to obtain a theobromine eluate.

An in vitro study on the metabolic interaction between ethanol and caffeine at high concentrations using human liver microsomes

Abstract: Caffeine (1,3,7-trimethylxanthine, CA) is a compound usually contained in beverages such as coffee and tea. CA is known to pharmacologically antagonize ethanol. Thus, the simultaneous intake of CA and ethanol takes place very commonly. Sometimes, toxic interactions also occur following combined intake of these two compounds. In this study, we conducted in vitro experiments to examine the interaction between ethanol and CA at high concentrations by observing mixed-function oxidation reaction using human liver microsomes. After incubation of the mixtures, CA and its three main metabolites theobromine (3,7-dimethylxanthine, TB), paraxanthine (1,7-dimethylxanthine, PX), and theophylline (1,3-dimethylxanthine, TP) were measured by high-performance liquid chromatography with ultraviolet detection. As results, the production of PX, the main metabolite of CA, was consistently inhibited by 24%–53% (P < 0.05) by high ethanol concentrations (20, 40, and 60 mM), but that of TP or TB was not. These results suggest that pharmacological or toxicological effects of CA may be enhanced by simultaneous use of ethanol and CA in humans.

Frozen confectionery products comprising theobromine and caffeine

Abstract: A frozen confection product is provided, the product comprising: from 20 to 80% of a frozen composition comprising cocoa solids; from 20 to 60% of a chocolate or chocolate analogue comprising cocoa solids; and from 5 to 20% (by weight of the product) of cocoa nibs; wherein the product contains at least 400 mg of theobromine and at least 40 mg of caffeine per 100 grams.

Production method and production device for composition having high content of theobromine

Abstract: An object is to provide a method for producing a high theobromine-containing composition effectively in a simple way. The method for producing a theobromine-containing composition comprises: (a)performing an extraction of a theobromine-containing plant or a processed product thereof with a solvent, to obtain a crude theobromine extract; (b) applying the crude theobromine extract to a cation exchange resin subjected in advance to hydrogen ion substitution, to allow an adsorption of theobromine onto the cation exchange resin; and (c) passing a solvent containing no ionic substance through the cation exchange resin subsequently to the step (b), to obtain a theobromine eluate.

progenies of Ilex paraguariensis St. Hil.

Abstract: Mate (Ilex paraguariensis St. Hil) contains phytochemical compounds capable of preventing a number of health problems. Knowledge on the genetic contribution to the variability in these compounds can help to obtain mate progenies with higher levels thereof in breeding programs. The composition of triterpene saponins, methylxanthines, chlorogenic acid and the antioxidant activity of eight mate progenies were evaluated. Significant differences among progenies were verified in contents of triterpene saponins (0.003-0.080%), caffeine (0.226-1.377%), theobromine (0.176-0.831%), and chlorogenic acid (1.344-2.031%) and in antioxidant activity (31.251-51.406%). The contents of theobromine were found to be negatively correlated with saponins and caffeine, and caffeine with chlorogenic acid, while theobromine was positively correlated with chlorogenic acid. The heritability values for saponins (75.09%), caffeine (75.19%), theobromine (66.87%), chlorogenic acid (52.86%) and antioxidant activity (67.75%) indicate the possibility of genetic gain in selection for these traits.