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Sample records for theobromine

  1. Interaction of theobromine with sodium benzoate

    Energy Technology Data Exchange (ETDEWEB)

    Nishijo, J.; Yonetani, I.

    1982-03-01

    The interaction of theobromine with sodium benzoate was investigated by PMR spectroscopy. The interaction of theobromine with pentadeuterated benzoic acid (benzoic acid-d5) was examined in the same manner but to a lesser degree. Chemical shifts of theobromine protons were determined as a function of sodium benzoate concentration in deuterium oxide at 30 and 15 degrees. Signals of both methyl groups of theobromine underwent significant upfield shifts when sodium benzoate was added to a theobromine solution. This fact suggests that a complex is formed by vertical stacking or plane-to-plane stacking. The same results were obtained for benzoic acid-d5.

  2. Theobromine induced seminiferous tubular lesion with elevated ...

    African Journals Online (AJOL)

    Concentrations of serum testosterone, albumin, transferrin, total serum protein and testicular histology of male albino wistar rats were evaluated following administration of 700mg/kg body weight theobromine in 0.5ml 14% sodium acetate for seven days. Theobromine administration produced significant increase in serum ...

  3. Theobromine inhibits sensory nerve activation and cough.

    Science.gov (United States)

    Usmani, Omar S; Belvisi, Maria G; Patel, Hema J; Crispino, Natascia; Birrell, Mark A; Korbonits, Márta; Korbonits, Dezso; Barnes, Peter J

    2005-02-01

    Cough is a common and protective reflex, but persistent coughing is debilitating and impairs quality of life. Antitussive treatment using opioids is limited by unacceptable side effects, and there is a great need for more effective remedies. The present study demonstrates that theobromine, a methylxanthine derivative present in cocoa, effectively inhibits citric acid-induced cough in guinea-pigs in vivo. Furthermore, in a randomized, double-blind, placebo-controlled study in man, theobromine suppresses capsaicin-induced cough with no adverse effects. We also demonstrate that theobromine directly inhibits capsaicin-induced sensory nerve depolarization of guinea-pig and human vagus nerve suggestive of an inhibitory effect on afferent nerve activation. These data indicate the actions of theobromine appear to be peripherally mediated. We conclude theobromine is a novel and promising treatment, which may form the basis for a new class of antitussive drugs.

  4. Remineralization of artificial enamel lesions by theobromine.

    Science.gov (United States)

    Amaechi, B T; Porteous, N; Ramalingam, K; Mensinkai, P K; Ccahuana Vasquez, R A; Sadeghpour, A; Nakamoto, T

    2013-01-01

    This study investigated the remineralization potential of theobromine in comparison to a standard NaF dentifrice. 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 theobromine (38 ± 32%) and toothpaste (29 ± 16%). With TMR (Δz/lD), theobromine and toothpaste exhibited significantly (p 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%). The present study demonstrated that theobromine in an apatite-forming medium can enhance the remineralization potential of the medium. Copyright © 2013 S. Karger AG, Basel.

  5. Salts and Co-crystals of Theobromine and their phase ...

    Indian Academy of Sciences (India)

    of stability, these salts transformed to theobromine within 1 h of dissolution in water. Remarkably, the besylate and tosylate salts are 88 and 58 times more soluble than theobromine, but they dissociated within 1 h. In contrast, theobromine co-crystals with gallic acid, anthranilic acid and 5-chlorosalicylic acid were found to be.

  6. Comparative effects of theobromine and Cocoa Bean Shell (CBS ...

    African Journals Online (AJOL)

    Synthetic theobromine and theobromine from CBS caused significant (p < 0.05) decrease in feed intake, body weight gain and significant (p < 0.05) increase in water intake relative to control. Theobromine also caused a significant (p < 0.05) increase in rectal temperature, pulse and respiratory rates. Serum parameters ...

  7. Psychopharmacology of theobromine in healthy volunteers.

    Science.gov (United States)

    Baggott, Matthew J; Childs, Emma; Hart, Amy B; de Bruin, Eveline; Palmer, Abraham A; Wilkinson, Joy E; de Wit, Harriet

    2013-07-01

    Theobromine, a methylxanthine related to caffeine and present in high levels in cocoa, may contribute to the appeal of chocolate. However, current evidence for this is limited. We conducted a within-subjects placebo-controlled study of a wide range of oral theobromine doses (250, 500, and 1,000 mg) using an active control dose of caffeine (200 mg) in 80 healthy participants. Caffeine had the expected effects on mood including feelings of alertness and cardiovascular parameters. Theobromine responses differed according to dose; it showed limited subjective effects at 250 mg and negative mood effects at higher doses. It also dose-dependently increased heart rate. In secondary analyses, we also examined individual differences in the drug's effects in relation to genes related to their target receptors, but few associations were detected. This study represents the highest dose of theobromine studied in humans. We conclude that theobromine at normal intake ranges may contribute to the positive effects of chocolate, but at higher intakes, effects become negative.

  8. Isolation and characterisation of theobromine-degrading filamentous fungi.

    Science.gov (United States)

    Oduro-Mensah, Daniel; Ocloo, Augustine; Lowor, Sammy T; Bonney, Evelyn Y; Okine, Laud K N A; Adamafio, Naa Ayikailey

    2018-01-01

    Strategies for achieving global food security include identification of alternative feedstock for use as animal feed, to contribute towards efforts at increasing livestock farming. The presence of theobromine in cocoa pod husks, a major agro-waste in cocoa-producing countries, hinders its utilisation for this purpose. Cheap treatment of cocoa pod husks to remove theobromine would allow largescale beneficial use of the millions of metric tonnes generated annually. The aim of this study was to isolate theobromine-degrading filamentous fungi that could serve as bioremediation agents for detheobromination of cocoa pod husks. Filamentous fungi were screened for ability to degrade theobromine. The most promising isolates were characterized with respect to optimal environmental conditions for theobromine degradation. Secretion of theobromine-degrading enzymes by the isolates was investigated. Theobromine degradation was monitored by HPLC. Of fourteen theobromine-degrading isolates collected and identified by rDNA 5.8S and ITS sequences, seven belonged to Aspergillus spp. and six were Talaromyces spp. Based on the extent of theobromine utilization, four isolates; Aspergillus niger, Talaromyces verruculosus and two Talaromyces marneffei, showed the best potential for use as bioagents for detheobromination. First-time evidence was found of the use of xanthine oxidase and theobromine oxidase in degradation of a methylxanthine by fungal isolates. Metabolism of theobromine involved initial demethylation at position 7 to form 3-methylxanthine, or initial oxidation at position 8 to form 3,7-dimethyuric acid. All four isolates degraded theobromine beyond uric acid. The data suggest that the four isolates can be applied to substrates, such as cocoa pod husks, for elimination of theobromine. Copyright © 2017 Elsevier GmbH. All rights reserved.

  9. Theobromine Effects on Enamel Surface Microhardness: In Vitro

    OpenAIRE

    Grace Syafira; Rina Permatasari; Nina Wardani

    2013-01-01

    Dental caries is still a dental health problem in Indonesia. Fluoride, one of the dental caries prevention material, but its safety and the danger of fluorosis is still debated. Theobromine is an alkaloid compound contained in cocoa beans. Theobromine is believed to increase enamel microhardness with mineral changes in the enamel superficial layer. Objectives: To determine the influence of theobromine on the enamel surface microhardness. Methods: This study used 40 premolar tooth crown pieces...

  10. Theobromine and the pharmacology of cocoa.

    Science.gov (United States)

    Smit, Hendrik Jan

    2011-01-01

    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.

  11. Plasma Lipid Profile and Target Organ Effect of Theobromine ...

    African Journals Online (AJOL)

    The influence of theobromine rich extracts of cocoa on plasma lipid profile and its organ specific effects on the kidney and liver were determined in twenty female weaning albino wistar rats. Varying concentration, 70 mg/kg, 140 mg/kg and 210 mg/kg body weight of theobromine in 0.9% normal saline were administered ...

  12. Effect Of Theobromine Exposure On Haematological Parameters In ...

    African Journals Online (AJOL)

    This work was carried out to investigate the effect of theobromine exposure on haematological indices in rats and thus evaluate whether its use in chemotherapy may be associated with possible side effects such as anaemia. Theobromine in two doses was administered by oral gavage to albino Wistar rats of both sexes (n ...

  13. Theobromine Effects on Enamel Surface Microhardness: In Vitro

    Directory of Open Access Journals (Sweden)

    Grace Syafira

    2013-07-01

    Full Text Available Dental caries is still a dental health problem in Indonesia. Fluoride, one of the dental caries prevention material, but its safety and the danger of fluorosis is still debated. Theobromine is an alkaloid compound contained in cocoa beans. Theobromine is believed to increase enamel microhardness with mineral changes in the enamel superficial layer. Objectives: To determine the influence of theobromine on the enamel surface microhardness. Methods: This study used 40 premolar tooth crown pieces that were embedded in epoxy resin. Furthermore specimens were randomly divided into 4 groups, which were control (distilled water, theobromine 100 mg/L (T100, theobromine 500 mg/L (T500 and theobromine 1000 mg/L (T1000. Specimens were immersed for 15 minutes and microhardness test was performed using Knoop microhardness tester. Results: Increasing enamel microhardness was observed after treatment with four different theobromine concentrations. The highest icreased of enamel microhardness was shown in T1000 group and difference compared to other groups were statistically significant (p<0.05. Conclusion: theobromine is a potential dental caries prevention material due to its effect in improving the microhardness of tooth enamel.

  14. Effects of theobromine and caffeine on mood and vigilance.

    Science.gov (United States)

    Judelson, Daniel A; Preston, Amy G; Miller, Debra L; Muñoz, Colleen X; Kellogg, Mark D; Lieberman, Harris R

    2013-08-01

    Like caffeine, theobromine crosses the blood-brain barrier and binds to adenosine receptors, suggesting it might share caffeine's beneficial effects on mood and vigilance. Therefore, the purpose of this study was to assess the effect of theobromine doses commonly found in foods on mood and vigilance parameters sensitive to caffeine. Caffeine was tested as a positive control. Twenty-four men (age, 23 [3] years) completed 6 double-blind trials during which they consumed experimental beverages, assessed their mood using standardized self-report questionnaires, and completed a 2-hour visual vigilance task. Three experimental doses (100, 200, and 400 mg theobromine) were delivered in a cocoa-based beverage; 3 matched control treatments (0 mg theobromine, 400 mg theobromine, and 100 mg caffeine) were delivered in a non-cocoa beverage. Mean salivary concentrations of theobromine exhibited significant dose-dependent differences (400 mg trials > 200 mg trial > 100 mg trial > 0 mg trials; P theobromine failed to consistently affect mood state or vigilance (P > 0.05), but 100-mg caffeine significantly decreased lethargy/fatigue and increased vigor (P = 0.006 and 0.011, respectively). These findings indicate theobromine does not influence mood and vigilance when administered in nutritionally relevant doses, despite sharing many of caffeine's structural characteristics.

  15. Theobromine enhances absorption of cacao polyphenol in rats

    National Research Council Canada - National Science Library

    Yamamoto, Takayuki; Takahashi, Haruya; Suzuki, Koichiro; Hirano, Akira; Kamei, Masanori; Goto, Tsuyoshi; Takahashi, Nobuyuki; Kawada, Teruo

    2014-01-01

    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...

  16. HPLC method for urinary theobromine determination: Effect of consumption of cocoa products on theobromine urinary excretion in children.

    Science.gov (United States)

    Rodriguez, Adrian; Costa-Bauza, Antonia; Saez-Torres, Concepcion; Rodrigo, Dolores; Grases, Felix

    2015-11-01

    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, was 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 Theobromine excretion levels were significantly higher in healthy children with higher consumption of cocoa products (ptheobromine determination with 100% recovery, without sample pretreatment. Urinary theobromine levels in healthy children were directly related to their consumption of cocoa products. Copyright © 2015 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  17. PENGARUH LEVEL DAN WAKTU MARINASI THEOBROMINE TERHADAP KUALITAS ORGANOLEPTIK DAGING SAPI BALI

    OpenAIRE

    ESARIANTO, ALIFRAN

    2016-01-01

    2015 ABSTRAK ALIFRAN ESARIANTO (I111 11 363). Pengaruh Level dan Waktu Marinasi Theobromine Terhadap Kualitas Organoleptik Daging Sapi Bali. Dibimbing oleh HIKMAH M. ALI dan ENDAH MURPININGRUM. Pemberian theobromine dapat meningkatkan kualitas organoleptik daging sapi Bali melalui perlakuan marinasi. Tujuan penelitian ini adalah untuk mengetahui pengaruh level dan lama marinasi theobromine serta interaksi keduanya terhadap kualitas organoleptik daging sapi Bali. Parameter yang diamat...

  18. Effect of cocoa and theobromine consumption on serum HDL-cholesterol concentrations: a randomized controlled trial.

    Science.gov (United States)

    Neufingerl, Nicole; Zebregs, Yvonne E M P; Schuring, Ewoud A H; Trautwein, Elke A

    2013-06-01

    Evidence from clinical studies has suggested that cocoa may increase high-density lipoprotein (HDL)-cholesterol concentrations. However, it is unclear whether this effect is attributable to flavonoids or theobromine, both of which are major cocoa components. We investigated whether pure theobromine increases serum HDL cholesterol and whether there is an interaction effect between theobromine and cocoa. The study had a 2-center, double-blind, randomized, placebo-controlled, full factorial parallel design. After a 2-wk run-in period, 152 healthy men and women (aged 40-70 y) were randomly allocated to consume one 200-mL drink/d for 4 wk that contained 1) cocoa, which naturally provided 150 mg theobromine and 325 mg flavonoids [cocoa intervention (CC)], 2) 850 mg pure theobromine [theobromine intervention (TB)], 3) cocoa and added theobromine, which provided 1000 mg theobromine and 325 mg flavonoids [theobromine and cocoa intervention (TB+CC)], or 4) neither cocoa nor theobromine (placebo). Blood lipids and apolipoproteins were measured at the start and end of interventions. In a 2-factor analysis, there was a significant main effect of the TB (P Theobromine independently increased serum HDL-cholesterol concentrations by 0.16 mmol/L. The lack of significant cocoa and interaction effects suggested that theobromine may be the main ingredient responsible for the HDL cholesterol-raising effect. This trial was registered at clinicaltrials.gov as NCT01481389.

  19. Effect of cocoa's theobromine on intestinal microbiota of rats.

    Science.gov (United States)

    Martín-Peláez, Sandra; Camps-Bossacoma, Mariona; Massot-Cladera, Malen; Rigo-Adrover, Mar; Franch, Àngels; Pérez-Cano, Francisco J; Castell, Margarida

    2017-10-01

    To establish the role of cocoa theobromine on gut microbiota composition and fermentation products after cocoa consumption in rats. Lewis rats were fed either a standard diet (RF diet), a diet containing 10% cocoa (CC diet) or a diet including 0.25% theobromine (TB diet) for 15 days. Gut microbiota (fluorescence in situ hybridization coupled to flow cytometry and metagenomics analysis), SCFA and IgA-coated bacteria were analyzed in fecal samples. CC and TB diets induced lower counts of E. coli whereas TB diet led to lower counts of Bifidobacterium spp., Streptococcus spp. and Clostridium histolyticum-C. perfingens group compared to RF diet. Metagenomics analysis also revealed a different microbiota pattern among the studied groups. The SCFA content was higher after both CC and TB diets, which was mainly due to enhanced butyric acid production. Furthermore, both diets decreased the proportion of IgA-coated bacteria. Cocoa's theobromine plays a relevant role in some effects related to cocoa intake, such as the lower proportion of IgA-coated bacteria. Moreover, theobromine modifies gut microbiota although other cocoa compounds could also act on intestinal bacteria, attenuating or enhancing the theobromine effects. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Maternal serum theobromine and the development of preeclampsia.

    Science.gov (United States)

    Klebanoff, Mark A; Zhang, Jun; Zhang, Cuilin; Levine, Richard J

    2009-09-01

    Preeclampsia, a disorder with prominent cardiovascular manifestations, is a cause of maternal, fetal, and infant morbidity and mortality. Chocolate contains compounds that may promote cardiovascular health. A recent study found chocolate consumption during pregnancy, and, particularly, increasing cord serum concentration of theobromine (the primary methylxanthine alkaloid in chocolate), to be associated with reduced occurrence of preeclampsia. We studied 2769 women who comprised the control group from a case-control study of caffeine metabolites and spontaneous abortion nested within the Collaborative Perinatal Project. These women were pregnant between 1959 and 1966, with liveborn infants of at least 28 weeks' gestation. Serum was drawn at 26 weeks' gestation, and assayed for theobromine by high-performance liquid chromatography. Odds ratios (ORs) for preeclampsia were estimated using logistic regression, and adjusted for age, education, prepregnant weight, race, parity, smoking, and gestation at blood draw. Preeclampsia occurred in 68 (2.9%) of 2105 eligible women. Adjusted ORs for preeclampsia were near unity across most third-trimester theobromine concentrations. Adjusted ORs for preeclampsia according to theobromine concentration in serum at theobromine and preeclampsia.

  1. comparative effects of theobromine and cocoa bean shell

    African Journals Online (AJOL)

    Mr. Oluyemi

    decrease in feed intake, body weight gain and significant (p < 0.05) increase in water intake relative to control. ... significant (p < 0.05) increase in these parameter compared with the control but lower than the effect from the synthetic theobromine. However ... weighed into a 500ml flat bottom flask and 150ml of water was.

  2. Supporting Information Salts and cocrystals of Theobromine and ...

    Indian Academy of Sciences (India)

    Supporting Information. Salts and cocrystals of Theobromine and their phase transformations in water. PALASH SANPHUI and ASHWINI NANGIA*. School of Chemistry, University of Hyderabad, Prof. C. R. Rao Road, Gachibowli, Central University P.O., Hyderabad 500 046, India. Table S1. Hydrogen bonds in crystal ...

  3. Biosynthesis of caffeine by tea-leaf extracts. Enzymic formation of theobromine from 7-methylxanthine and of caffeine from theobromine.

    Science.gov (United States)

    Suzuki, T; Takahashi, E

    1975-01-01

    1. Extracts prepared from tea leaves with Polyclar AT (insoluble polyvinylpyrrolidine) contained two methyltransferase activities catalysing the transfer of methyl groups from S-adenosylmethionine to 7-methylxanthine, producing theobromine, and to theobromine, producing caffeine. 2. The methyltransferases exhibited the same pH optimum (8.4) and a similar pattern of effects by metal ions, thiol inhibitors and metal-chelating reagents, both for theobromine and caffeine synthesis. Mg2+, Mn2+ and Ca2+ slightly stimulated enzyme activity but they were not essential. Paraxanthine was shown to be most active among methylxanthines, as the methyl acceptor. However, the formation of paraxanthine from 1-methylxanthine was very low and that from 7-methylxanthine was nil, suggesting that the synthesis of caffeine from paraxanthine is of little importance in intact plants. Xanthine, xanthosine, XMP and hypoxanthine were all inactive as methyl acceptors, whereas [2(-14)C]xanthine and [8(-14)C]hypoxanthine were catabolized to allantoin and urea by tea-leaf extracts. The apparent Km values are as follows: 7-methylxanthine, 1.0 times 10(-14)M; theobromine, 1.0 times 10(-3)M; paraxanthine, 0.2 times 10(-3)M; S-adenosylmethionine, 0.25 times 10(-4)M (with each of the three substrates). 3. The results suggest that the pathway for caffeine biosynthesis is as follows: 7-methylxanthine leads to theobromine leads to caffeine. In contrast, it is suggested that theophylline is synthesized from 1-methylxanthine. The methyl groups of the purine ring of caffeine are all derived directly from the methyl group of S-adenosylmethionine. Little is known about the pathways leading to the formation of 7-methylxanthine. 4. A good correlation between caffeine synthesis and shoot formation or growth of tea seedlings was shown, suggesting that the methylating systems in caffeine synthesis are closely associated with purine nucleotide and nucleic acid metabolism in tea plants. PMID:238504

  4. Theobromine suppresses adipogenesis through enhancement of CCAAT-enhancer-binding protein β degradation by adenosine receptor A1.

    Science.gov (United States)

    Mitani, Takakazu; Watanabe, Shun; Yoshioka, Yasukiyo; Katayama, Shigeru; Nakamura, Soichiro; Ashida, Hitoshi

    2017-12-01

    Theobromine, a methylxanthine derived from cacao beans, reportedly has various health-promoting properties but molecular mechanism by which effects of theobromine on adipocyte differentiation and adipogenesis remains unclear. In this study, we aimed to clarify the molecular mechanisms of the anti-adipogenic effect of theobromine in vitro and in vivo. ICR mice (4week-old) were administered with theobromine (0.1g/kg) for 7days. Theobromine administration attenuated gains in body and epididymal adipose tissue weights in mice and suppressed expression of adipogenic-associated genes in mouse adipose tissue. In 3T3-L1 preadipocytes, theobromine caused degradation of C/EBPβ protein by the ubiquitin-proteasome pathway. Pull down assay showed that theobromine selectively interacts with adenosine receptor A1 (AR1), and AR1 knockdown inhibited theobromine-induced C/EBPβ degradation. Theobromine increased sumoylation of C/EBPβ at Lys133. Expression of the small ubiquitin-like modifier (SUMO)-specific protease 2 (SENP2) gene, coding for a desumoylation enzyme, was suppressed by theobromine. In vivo knockdown studies showed that AR1 knockdown in mice attenuated the anti-adipogenic effects of theobromine in younger mice. Theobromine suppresses adipocyte differentiation and induced C/EBPβ degradation by increasing its sumoylation. Furthermore, the inhibition of AR1 signaling is important for theobromine-induced C/EBPβ degradation. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Stimulation of Clostridium perfringens enterotoxin formation by caffeine and theobromine.

    Science.gov (United States)

    Labbe, R G; Nolan, L L

    1981-01-01

    In the presence of 100 micrograms of caffeine per ml or 200 micrograms of theobromine per ml, sporulation of Clostridium perfringens NCTC 8679 rose from less than 1 to 80 or 85%. Enterotoxin concentration increased from undetectable levels to 450 micrograms/mg of cell extract protein. Heat-resistant spore levels increased from less than 1,000 to between 1 X 10(7) and 2 X 10(7)/ml. These effects were partially reversible by the addition of adenosine or thymidine. In the case of NCTC 8238, caffeine and theobromine caused a three- to fourfold increase in the percentages of cells possessing refractile spores and a similar increase in enterotoxin concentration. Heat-resistant spore levels, however, were unaffected. Inosine was ineffective in promoting sporulation in NCTC 8679. PMID:6271685

  6. Theobromine enhances absorption of cacao polyphenol in rats.

    Science.gov (United States)

    Yamamoto, Takayuki; Takahashi, Haruya; Suzuki, Koichiro; Hirano, Akira; Kamei, Masanori; Goto, Tsuyoshi; Takahashi, Nobuyuki; Kawada, Teruo

    2014-01-01

    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.

  7. Metabolic engineering of Saccharomyces cerevisiae for caffeine and theobromine production.

    Directory of Open Access Journals (Sweden)

    Lu Jin

    Full Text Available Caffeine (1, 3, 7-trimethylxanthine and theobromine (3, 7-dimethylxanthine are the major purine alkaloids in plants, e.g., tea (Camellia sinensis and coffee (Coffea arabica. Caffeine is a major component of coffee and is used widely in food and beverage industries. Most of the enzymes involved in the caffeine biosynthetic pathway have been reported previously. Here, we demonstrated the biosynthesis of caffeine (0.38 mg/L by co-expression of Coffea arabica xanthosine methyltransferase (CaXMT and Camellia sinensis caffeine synthase (TCS in Saccharomyces cerevisiae. Furthermore, we endeavored to develop this production platform for making other purine-based alkaloids. To increase the catalytic activity of TCS in an effort to increase theobromine production, we identified four amino acid residues based on structural analyses of 3D-model of TCS. Two TCS1 mutants (Val317Met and Phe217Trp slightly increased in theobromine accumulation and simultaneously decreased in caffeine production. The application and further optimization of this biosynthetic platform are discussed.

  8. Reinforcing effects of caffeine and theobromine as found in chocolate.

    Science.gov (United States)

    Smit, Hendrik J; Blackburn, Rachel J

    2005-08-01

    Although in a previous study we showed that caffeine and theobromine were the main psychopharmacologically active constituents in a 50-g bar of chocolate, mere activity does not guarantee a role in our liking for the food. Our aim was to see if liking for a drink repeatedly paired with these amounts of caffeine and theobromine would increase compared to a placebo-paired drink. Participants (n=64) consumed a 'novel' drink + treatment capsule on six non-consecutive mornings using a double-blind, placebo-controlled independent-sample design. Aspects of liking and intensity of various sensory descriptors for these drinks were measured at every drink collection. Treatment capsules contained either an ecologically relevant dose combination of 19-mg caffeine and 250-mg theobromine or a placebo. Liking for the drink paired with the methylxanthine-containing capsules increased over time compared to the placebo-paired drink. This highly significant effect was confirmed by subjective, retrospective changes in liking for the drink. Methylxanthines in amounts found in 50-g chocolate may well contribute to our liking for chocolate, especially to the more acquired taste for dark chocolate.

  9. Antioxidant and prooxidant properties of caffeine, theobromine and xanthine.

    Science.gov (United States)

    Azam, Sonish; Hadi, Naghma; Khan, Nizam Uddin; Hadi, Sheikh Mumtaz

    2003-09-01

    Caffeine, along with its catabolic products theobromine and xanthine, is a key component of tea and coffee. These compounds are structurally similar to uric acid, a known antioxidant which is present in blood at relatively high concentrations, but also shows prooxidant activity. In view of the structural similarity between uric acid and caffeine and its metabolites, we studied the antioxidant and prooxidant properties of these compounds. Antioxidant activity was determined by measuring the quenching effect of the compounds on oxidative DNA degradation by a hydroxyl radical generating system. Prooxidant activity was studied by measuring the ability of the compounds to oxidatively degrade DNA in the presence of copper ions. Caffeine, theobromine and xanthine have a quenching effect on the production of hydroxyl radicals, as well as on oxidative DNA breakage by hydroxyl radicals. Consistent with previous observations that many known antioxidants of plant origin are also capable of prooxidant action, the purine alkaloids also show oxidative DNA breakage in the presence of transition metal ions. The alkaloid caffeine and its catabolic products theobromine and xanthine exhibit both antioxidant and prooxidant properties. The results lead to the observation that caffeine and its metabolites may also contribute to the overall antioxidant and chemopreventive properties of caffeine-bearing beverages, such as tea.

  10. Metabolic engineering of Saccharomyces cerevisiae for caffeine and theobromine production.

    Science.gov (United States)

    Jin, Lu; Bhuiya, Mohammad Wadud; Li, Mengmeng; Liu, XiangQi; Han, Jixiang; Deng, WeiWei; Wang, Min; Yu, Oliver; Zhang, Zhengzhu

    2014-01-01

    Caffeine (1, 3, 7-trimethylxanthine) and theobromine (3, 7-dimethylxanthine) are the major purine alkaloids in plants, e.g., tea (Camellia sinensis) and coffee (Coffea arabica). Caffeine is a major component of coffee and is used widely in food and beverage industries. Most of the enzymes involved in the caffeine biosynthetic pathway have been reported previously. Here, we demonstrated the biosynthesis of caffeine (0.38 mg/L) by co-expression of Coffea arabica xanthosine methyltransferase (CaXMT) and Camellia sinensis caffeine synthase (TCS) in Saccharomyces cerevisiae. Furthermore, we endeavored to develop this production platform for making other purine-based alkaloids. To increase the catalytic activity of TCS in an effort to increase theobromine production, we identified four amino acid residues based on structural analyses of 3D-model of TCS. Two TCS1 mutants (Val317Met and Phe217Trp) slightly increased in theobromine accumulation and simultaneously decreased in caffeine production. The application and further optimization of this biosynthetic platform are discussed.

  11. Theobromine Inhibits Uric Acid Crystallization. A Potential Application in the Treatment of Uric Acid Nephrolithiasis

    Science.gov (United States)

    Grases, Felix; Rodriguez, Adrian; Costa-Bauza, Antonia

    2014-01-01

    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. PMID:25333633

  12. Theobromine inhibits uric acid crystallization. A potential application in the treatment of uric acid nephrolithiasis.

    Science.gov (United States)

    Grases, Felix; Rodriguez, Adrian; Costa-Bauza, Antonia

    2014-01-01

    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. 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. 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. 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.

  13. Theobromine inhibits uric acid crystallization. A potential application in the treatment of uric acid nephrolithiasis.

    Directory of Open Access Journals (Sweden)

    Felix Grases

    Full Text Available 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.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.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.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.

  14. RNA binding efficacy of theophylline, theobromine and caffeine.

    Science.gov (United States)

    Johnson, I Maria; Kumar, S G Bhuvan; Malathi, R

    2003-04-01

    The binding of naturally occurring methylxanthines such as theophylline, theobromine and caffeine to nucleic acids are reckoned to be pivotal as they are able to modulate the cellular activities. We explore the interaction of yeast RNA binding efficacy of the above xanthine derivatives by using UV absorption differential spectroscopy and Fourier Transform Infrared (FTIR) spectroscopy. Both the analyses show discrimination in their binding affinity to RNA. The differential UV-spectrum at P/D 3.3 reveals the greater RNA binding activity for theophylline (85 +/- 5%), whereas moderate and comparatively less binding activity for theobromine (45 +/- 5%) and caffeine (30 +/- 5%) and the binding activity was found to depend on concentration of the drugs. In FTIR analysis we observed changes in the amino group (NH) of RNA complexed by drugs, where the NH band is found to become very broad, indicating hydrogen bonding (H-bonding) with theophylline (3343.4 cm(-1)), theobromine (3379.8 cm(-1)) and caffeine (3343 cm(-1)) as compared to the free RNA (3341.6 cm(-1)). Furthermore in RNA-theophylline complex, it is observed that the carbonyl (C=O) vibration frequency (nu(C=O)) of both drug (nu(C=O)=1718, 1666 cm(-1)) as well as RNA (nu(C=O)=1699, 1658 cm(-1)) disappeared and a new vibration band appeared around 1703 cm(-1), indicating that the C=O and NH groups of drug and RNA are effectively involved in H-bonding. Whereas in RNA-theobromine and RNA-caffeine complexes, we found very little changes in C=O frequency and only broadening of the NH band of RNA due to complexation is observed in these groups. The changes in the vibrations of G-C/A-U bands and other bending frequencies are discussed. Thus the discrimination in the binding affinity of methylxanthines with RNA molecule shows that strong RNA binding drugs like theophylline can selectively be delivered to RNA targets of microbial pathogens having the mechanism of RNA catalysis.

  15. Structure of 2,4-Diaminopyrimidine - Theobromine Alternate Base Pairs

    Science.gov (United States)

    Gengeliczki, Zsolt; Callahan, Michael P.; Kabelac, Martin; Rijs, Anouk M.; deVries, Mattanjah S.

    2011-01-01

    We report the structure of clusters of 2,4-diaminopyrimidine with 3,7-dimethylxanthine (theobromine) in the gas phase determined by IR-UV double resonance spectroscopy in both the near-IR and mid-IR regions in combination with ab initio computations. These clusters represent potential alternate nucleobase pairs, geometrically equivalent to guanine-cytosine. We have found the four lowest energy structures, which include the Watson-Crick base pairing motif. This Watson-Crick structure has not been observed by resonant two-photon ionization (R2PI) in the gas phase for the canonical DNA base pairs.

  16. The effects of fluoride, strontium, theobromine and their combinations on caries lesion rehardening and fluoridation.

    Science.gov (United States)

    Lippert, Frank

    2017-08-01

    The aim was to investigate the effects of fluoride, strontium, theobromine and their combinations on caries lesion rehardening and fluoridation (EFU) under pH cycling conditions. Human enamel specimens were demineralized at 37°C for 24h using a pH 5.0 solution containing 50mM lactic acid and 0.2% Carbopol 907 which was 50% saturated with respect to hydroxyapatite. Lesions were assigned to nine treatment groups (n=16) based on Knoop surface microhardness indentation length. aqueous solutions were: placebo, 11.9mM sodium fluoride (F), 23.8mM sodium fluoride (2×F), 1.1mM strontium chloride hexahydrate (Sr), 1.1mM F theobromine, Sr+theobromine, F+Sr, F+theobromine, F+Sr+theobromine. Lesions were pH cycled for 5d (daily protocol: 3×1min-treatment; 2×60min-demineralization; 4×60min & overnight-artificial saliva). Knoop indentation length was measured again and%surface microhardness recovery (%SMHr) calculated. EFU was determined using the acid-etch technique. Data were analysed using ANOVA. Model showed fluoride dose-response for both variables (2×F>F>placebo). For%SMHr, F+Sr+/-theobromine resulted in more rehardening than F, however less than 2×F. F+theobromine was similar to F. For EFU, F+Sr was inferior to F, F+theobromine and F+Sr+theobromine which were similar and inferior to 2×F. In absence of fluoride, Sr, theobromine or Sr+theobromine were virtually indistinguishable from placebo and inferior to F. It can be concluded that a) strontium aids rehardening but not EFU and only in presence of fluoride; b) theobromine does not appear to offer any anti-caries benefits in this model; c) there are no synergistic effects between strontium and theobromine in the presence or absence of fluoride. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Differential contributions of theobromine and caffeine on mood, psychomotor performance and blood pressure.

    Science.gov (United States)

    Mitchell, E S; Slettenaar, M; vd Meer, N; Transler, C; Jans, L; Quadt, F; Berry, M

    2011-10-24

    The combination of theobromine and caffeine, methylxanthines found in chocolate, has previously been shown to improve mood and cognition. However, it is unknown whether these molecules act synergistically. This study tested the hypothesis that a combination of caffeine and theobromine has synergistic effects on cognition, mood and blood pressure in 24 healthy female subjects. The effects of theobromine (700 mg), caffeine (120 mg) or the combination of both, or placebo were tested on mood (the Bond-Lader visual analog scale), psychomotor performance (the Digit Symbol Substitution Test (DSST)) and blood pressure before and at 1, 2 and 3 h after administration. Theobromine alone decreased self-reported calmness 3h after ingestion and lowered blood pressure relative to placebo 1 h after ingestion. Caffeine increased self-reported alertness 1, 2 and 3h after ingestion and contentedness 1 and 2 h after ingestion, and increased blood pressure relative to placebo (at 1 h). The combination of caffeine+theobromine had similar effects as caffeine alone on mood, but with no effect on blood pressure. There was no treatment effect on DSST performance. Together these results suggest that theobromine and caffeine could have differential effects on mood and blood pressure. It was tentatively concluded that caffeine may have more CNS-mediated effects on alertness, while theobromine may be acting primarily via peripheral physiological changes. Copyright © 2011 Elsevier Inc. All rights reserved.

  18. Theobromine intoxication in a red fox and a European badger in Sweden.

    Science.gov (United States)

    Jansson, D S; Galgan, V; Schubert, B; Segerstad, C H

    2001-04-01

    A red fox (Vulpes vulpes) and a European badger (Meles meles) were found dead on a golf-course in October 1997 near Stockholm (Sweden). At necropsy, both animals were obese and the main finding was acute circulatory collapse. Theobromine intoxication was suspected as chocolate waste was available at a nearby farm and no other cause of death could be detected. Gastric contents and samples of liver from both animals were analyzed by reversed-phase high pressure liquid chromatography for the presence of methylxanthines. Theobromine and caffeine were detected in gastric contents and theobromine was identified in the liver samples from both animals. This appears to be the first report of theobromine intoxication in the red fox and the European badger.

  19. Theobromine increases NAD⁺/Sirt-1 activity and protects the kidney under diabetic conditions.

    Science.gov (United States)

    Papadimitriou, Alexandros; Silva, Kamila C; Peixoto, Elisa B M I; Borges, Cynthia M; Lopes de Faria, Jacqueline M; Lopes de Faria, José B

    2015-02-01

    Reduction in sirtuin 1 (Sirt-1) is associated with extracellular matrix (ECM) accumulation in the diabetic kidney. Theobromine may reduce kidney ECM accumulation in diabetic rats. In the current study, we aimed to unravel, under diabetic conditions, the mechanism of kidney ECM accumulation induced by a reduction in Sirt-1 and the effect of theobromine in these events. In vitro, we used immortalized human mesangial cells (iHMCs) exposed to high glucose (HG; 30 mM), with or without small interfering RNA for NOX4 and Sirt-1. In vivo, spontaneously hypertensive rats (SHR) were rendered diabetic by means of streptozotocin and studied after 12 wk. The effects of treatment with theobromine were investigated under both conditions. HG leads to a decrease in Sirt-1 activity and NAD(+) levels in iHMCs. Sirt-1 activity could be reestablished by treatment with NAD(+), silencing NOX4, and poly (ADP-ribose) polymerase-1 (PARP-1) blockade, or with theobromine. HG also leads to a low AMP/ATP ratio, acetylation of SMAD3, and increased collagen IV, which is prevented by theobromine. Sirt-1 or AMPK blockade abolished these effects of theobromine. In diabetic SHR, theobromine prevented increases in albuminuria and kidney collagen IV, reduced AMPK, elevated NADPH oxidase activity and PARP-1, and reduced NAD(+) levels and Sirt-1 activity. These results suggest that in diabetes mellitus, Sirt-1 activity is reduced by PARP-1 activation and NAD(+) depletion due to low AMPK, which increases NOX4 expression, leading to ECM accumulation mediated by transforming growth factor (TGF)-β1 signaling. It is suggested that Sirt-1 activation by theobromine may have therapeutic potential for diabetic nephropathy. Copyright © 2015 the American Physiological Society.

  20. Theobromine up-regulates cerebral brain-derived neurotrophic factor and facilitates motor learning in mice.

    Science.gov (United States)

    Yoneda, Mitsugu; Sugimoto, Naotoshi; Katakura, Masanori; Matsuzaki, Kentaro; Tanigami, Hayate; Yachie, Akihiro; Ohno-Shosaku, Takako; Shido, Osamu

    2017-01-01

    Theobromine, which is a caffeine derivative, is the primary methylxanthine produced by Theobroma cacao. Theobromine works as a phosphodiesterase (PDE) inhibitor to increase intracellular cyclic adenosine monophosphate (cAMP). cAMP activates the cAMP-response element-binding protein (CREB), which is involved in a large variety of brain processes, including the induction of the brain-derived neurotrophic factor (BDNF). BDNF supports cell survival and neuronal functions, including learning and memory. Thus, cAMP/CREB/BDNF pathways play an important role in learning and memory. Here, we investigated whether orally administered theobromine could act as a PDE inhibitor centrally and affect cAMP/CREB/BDNF pathways and learning behavior in mice. The mice were divided into two groups. The control group (CN) was fed a normal diet, whereas the theobromine group (TB) was fed a diet supplemented with 0.05% theobromine for 30 days. We measured the levels of theobromine, phosphorylated vasodilator-stimulated phosphoprotein (p-VASP), phosphorylated CREB (p-CREB), and BDNF in the brain. p-VASP was used as an index of cAMP increases. Moreover, we analyzed the performance of the mice on a three-lever motor learning task. Theobromine was detectable in the brains of TB mice. The brain levels of p-VASP, p-CREB, and BDNF were higher in the TB mice compared with those in the CN mice. In addition, the TB mice performed better on the three-lever task than the CN mice did. These results strongly suggested that orally administered theobromine acted as a PDE inhibitor in the brain, and it augmented the cAMP/CREB/BDNF pathways and motor learning in mice. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Theobromine Inhibits Uric Acid Crystallization. A Potential Application in the Treatment of Uric Acid Nephrolithiasis

    OpenAIRE

    Felix Grases; Adrian Rodriguez; Antonia Costa-Bauza

    2014-01-01

    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 theobr...

  2. Biosynthesis, accumulation and degradation of theobromine in developing Theobroma cacao fruits.

    Science.gov (United States)

    Zheng, Xin-Qiang; Koyama, Yoko; Nagai, Chifumi; Ashihara, Hiroshi

    2004-04-01

    We have studied the purine alkaloid content and purine metabolism in Theobroma cacao fruits at differing growth stages: Stage A (young small fruit, fresh weight, ca. 2 g); stage B (medium size fruit, fresh weight, ca. 100 g) and stage C (large size, fresh weight, ca. 500 g). The major purine alkaloid in stage A fruits (mainly pericarp) was theobromine (0.7 micromol g(-1) fresh weight), followed by caffeine (0.09 micromol g(-1) fresh weight). The theobromine content of the pericarp decreased sharply with tissue age, and the caffeine content decreased gradually. A large amount of theobromine (22 micromol g(-1) fresh weight) had accumulated in seeds (mainly cotyledons) of stage C fruits. Theobromine was found also in the seed coat and placenta. Tracer experiments with [8-(14)C]adenine show that the major sites of theobromine synthesis are the young pericarp and cotyledons of T. cacao fruits. Limited amounts of purine alkaloids may be transported from the pericarp to seed tissue, but most purine alkaloids that accumulated in seeds appeared to be synthesised in cotyledons. Degradation of [8-(14)C]theobromine and [8-(14)C]caffeine to CO2 via 3-methylxanthine and ureides (allantoin and allantoic acid) was detected only in the pericarp of stage C fruits.

  3. Effects on peripheral and central blood pressure of cocoa with natural or high-dose theobromine: A randomized, double-blind crossover trial

    National Research Council Canada - National Science Library

    Bogaard, Bas; ijer, Richard; Westerhof, Berend; Meiracker, Anton; Montfrans, Gert; Born, B.J.H

    2010-01-01

    .... It has been suggested that theobromine is partially responsible for this effect. We tested whether consumption of flavanol-rich cocoa drinks with natural or added theobromine could lower peripheral and central blood pressure...

  4. Evaluation of human enamel surfaces treated with theobromine: a pilot study.

    Science.gov (United States)

    Kargul, Betul; Özcan, Mutlu; Peker, Sertac; Nakamoto, Tetsuo; Simmons, William B; Falster, Alexander U

    2012-01-01

    The objectives of this in-vitro study were to investigate the effect of theobromine, which is the principle xanthine species in Theobroma cacao, at two concentrations on the surface hardness and topography of human enamel. Twenty-four freshly extracted human third molars were collected and stored in distilled water with 0.1% thymol solution at room temperature prior to the experiments. The enamel specimens were treated with one coat of theobromine at two concentrations (100 mg/l or 200 mg/l in distilled water) for 5 min. Enamel surfaces in the control group received no theobromine. They were then kept in distilled water for 1 week and subjected to SEM analysis. The specimens were demineralised by storing them in acidic hydroxyethylcellulose for three days. After baseline microhardness measurements, they were incubated either in 100 or 200 mg/l theobromine for 5 min. The control group was kept in distilled water. After washing the specimens under distilled water, they were kept in a remineralising solution for 18 h. Microhardness of the enamel surface was initially determined for each specimen before artificial demineralisation. After demineralisation, the experimental groups were incubated in 100 mg or 200 mg theobromine and control-group specimens were placed in remineralising solution. Enamel surfaces of the untreated control group presented a generally smooth and slightly hummocky surface with small lines of pits. Specimens treated with theobromine showed differences between the two concentrations. The group treated with 200 mg/l solution for 5 min showed a greater quantity of globules on enamel than did specimens treated with 100 mg/l solution. As shown by the microhardness values, a consistent and remarkable protection of the enamel surface was found with the application of theobromine.

  5. NILAI CERNA DAN BIODEGRADASI THEOBROMIN POD KAKAO DENGAN PERLAKUAN FERMENTASI MENGGUNAKAN INOKULUM MULTI MIKROBIA (Digestibility Value and Theobromine Biodegradation of Cocoa Pod with Treatment of Fermentation Using Multi Microbial Inoculum

    Directory of Open Access Journals (Sweden)

    Suci Wulandari

    2014-07-01

    SBP terhadap nilai cerna serat dan biodegradasi theobromin. Pod kakao difermentasi dalam kondisi anaerob pada suhu ruang. Kadar inokulum yang ditambahkan adalah 0; 0,05; dan 0,1 %. Selama fermentasi dilakukan pengambilan sampel pada hari ke-0, 3, dan 6 untuk mengetahui pertumbuhan bakteri selama fermentasi pod kakao, perubahan pH, perubahan nilai cerna, dan degradasi theobromin selama fermentasi, kemudian dilanjutkan dengan isolasi bakteri untuk mengetahui karakteristik isolat bakteri dari pod kakao terfermentasi dengan cara isolat yang diperoleh diuji kemampuannya dalam menghasilkan enzim selulase (CMC-ase dan β-glukoseidase dan kemampuannya dalam mendegradasi theobromin. Hasil penelitian menunjukkan bahwa selama fermentasi pod kakao terjadi kenaikan populasi bakteri asam laktat dan bakteri total, penurunan pH, kenaikan nilai cerna serat, dan penurunan kandungan theobromin dalam pod kakao. Perubahan parameter tersebut lebih nyata terjadi pada pod kakao yang difermentasi dengan kadar air 40% dan diinokulasi dengan SBP® dosis 0,05% dengan lama fermentasi 6 hari. Penurunan theobromin pada pod kakao fermentasi terbesar mencapai 17,02%. Kenaikan nilai cerna serat dan penurunan kandungan theobromin selama fermentasi pod kakao oleh inokulan SBP dikonfirmasi dengan karakteristik isolat-isolat bakteri yang diperoleh dari pod kakao yang terfermentasi. Ada 8 isolat bakteri yang mampu menghasilkan enzim CMC-ase dan β-glukosidase dengan tingkat produksi yang berbeda-beda. Beberapa isolat bakteri juga mampu menurunkan kandungan theobromin dalam kultur cair sampai sebesar 27,07%. Kata kunci: Pod kakao, theobromin, nilai cerna serat, selulase

  6. Proximate Composition, Extraction, and Purification of Theobromine from Cacao Pod Husk (Theobroma Cacao L.

    Directory of Open Access Journals (Sweden)

    Van Tang Nguyen

    2017-04-01

    Full Text Available The aims of this study were to determine the proximate composition of cacao pod husk as well as the optimal conditions for extraction and purification of theobromine from cacao pod husk. The results indicated that cacao pod husk had high contents of moisture and carbohydrate (87.06% and 11.03% by fresh weight, respectively, but low contents of crude protein, crude lipid, and ash (0.31%, 0.12%, and 1.48% by fresh weight, respectively. The optimal conditions for extraction of theobromine from cacao pod husk were of 70% ethanol, with an extraction time of 90 min, and 1 as the number of extractions. A concentration of 10% by volume of 10% lead acetate solution was the best selection for purification of the crude extracts containing theobromine from cacao pod husk. Under these optimal conditions, theobromine content obtained from cacao pod husk was 6.79 mg/100 g dry weight. The finding from this study is a valuable contribution for obtaining theobromine from an abundant, inexpensive, renewable, and sustainable source for potential application in the nutraceutical, medical, and pharmaceutical industries.

  7. The relevance of theobromine for the beneficial effects of cocoa consumption

    Directory of Open Access Journals (Sweden)

    Eva eMartínez-Pinilla

    2015-02-01

    Full Text Available Cocoa consumption began in America and in the mid 16th Century it quickly spread to Europe. Beyond being considered a pleasant habit due to its rich sweet lingering taste, chocolate was considered a good nutrient and even a medicine. Traditionally, health benefits of cocoa have been related with the high content of antioxidants of Theobroma cocoa beans. However, the direct psychoactive effect due to methylxanthines in cocoa is notable. Theobromine and caffeine in the proportions found in cocoa are responsible for the liking of the food/beverage. These compounds influence in a positive way our moods and our state of alertness. Theobromine, which is found in higher amounts than caffeine, seems to be behind several effects attributed to cocoa intake. The main mechanisms of action are inhibition of phosphodiesterases and blockade of adenosine receptors. Further mechanisms are being explored to better understand the health benefits associated to theobromine consumption. Unlike what happens in other mammals –pets included-, theobromine is safe for humans and has fewer unwanted effects than caffeine. Therefore, theobromine deserves attention as one of the most attractive molecules in cocoa.

  8. The relevance of theobromine for the beneficial effects of cocoa consumption.

    Science.gov (United States)

    Martínez-Pinilla, Eva; Oñatibia-Astibia, Ainhoa; Franco, Rafael

    2015-01-01

    Cocoa consumption began in America and in the mid sixteenth Century it quickly spread to Europe. Beyond being considered a pleasant habit due to its rich sweet lingering taste, chocolate was considered a good nutrient and even a medicine. Traditionally, health benefits of cocoa have been related with the high content of antioxidants of Theobroma cocoa beans. However, the direct psychoactive effect due to methylxanthines in cocoa is notable. Theobromine and caffeine, in the proportions found in cocoa, are responsible for the liking of the food/beverage. These compounds influence in a positive way our moods and our state of alertness. Theobromine, which is found in higher amounts than caffeine, seems to be behind several effects attributed to cocoa intake. The main mechanisms of action are inhibition of phosphodiesterases and blockade of adenosine receptors. Further mechanisms are being explored to better understand the health benefits associated to theobromine consumption. Unlike what happens in other mammals -pets- included, theobromine is safe for humans and has fewer unwanted effects than caffeine. Therefore, theobromine deserves attention as one of the most attractive molecules in cocoa.

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

    Science.gov (United States)

    Ishida, Mariko; Kitao, Naoko; Mizuno, Kouichi; Tanikawa, Natsu; Kato, Misako

    2009-02-01

    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.

  10. Proximate Composition, Extraction, and Purification of Theobromine from Cacao Pod Husk (Theobroma Cacao L.)

    OpenAIRE

    Van Tang Nguyen; Nghia Huu Nguyen

    2017-01-01

    The aims of this study were to determine the proximate composition of cacao pod husk as well as the optimal conditions for extraction and purification of theobromine from cacao pod husk. The results indicated that cacao pod husk had high contents of moisture and carbohydrate (87.06% and 11.03% by fresh weight, respectively), but low contents of crude protein, crude lipid, and ash (0.31%, 0.12%, and 1.48% by fresh weight, respectively). The optimal conditions for extraction of theobromine from...

  11. Caffeine and theobromine in epicuticular wax of Ilex paraguariensis A. St.-Hil.

    Science.gov (United States)

    Athayde, M L; Coelho, G C; Schenkel, E P

    2000-12-01

    Caffeine and theobromine were identified and quantified in leaf epicuticular waxes of Ilex paraguariensis A. St.-Hil. (Aquifoliaceae). The total epicuticular leaf wax content was ca. 0.5% on average of dry leaf weight. Epicuticular caffeine and theobromine contents varied from 0.16 to 127.6 microg/mg and from 0 to 9.5 microg/mg of wax, respectively. For some selected samples, the intracellular methylxanthine concentration was also determined. A positive correlation was found between inner and epicuticular caffeine contents.

  12. Cytochrome P450 isoform selectivity in human hepatic theobromine metabolism

    Science.gov (United States)

    Gates, Simon; Miners, John O

    1999-01-01

    Aims The plasma clearance of theobromine (TB; 3,7-dimethylxanthine) is known to be induced in cigarette smokers. To determine whether TB may serve as a model substrate for cytochrome P450 (CYP) 1A2, or possibly other isoforms, studies were undertaken to identify the individual human liver microsomal CYP isoforms responsible for the conversion of TB to its primary metabolites. Methods The kinetics of formation of the primary TB metabolites 3-methylxanthine (3-MX), 7-methylxanthine (7-MX) and 3,7-dimethyluric acid (3,7-DMU) by human liver microsomes were characterized using a specific hplc procedure. Effects of CYP isoform-selective xenobiotic inhibitor/substrate probes on each pathway were determined and confirmatory studies with recombinant enzymes were performed to define the contribution of individual isoforms to 3-MX, 7-MX and 3,7-DMU formation. Results The CYP1A2 inhibitor furafylline variably inhibited (0–65%) 7-MX formation, but had no effect on other pathways. Diethyldithiocarbamate and 4-nitrophenol, probes for CYP2E1, inhibited the formation of 3-MX, 7-MX and 3,7-DMU by ≈55–60%, 35–55% and 85%, respectively. Consistent with the microsomal studies, recombinant CYP1A2 and CYP2E1 exhibited similar apparent Km values for 7-MX formation and CYP2E1 was further shown to have the capacity to convert TB to both 3-MX and 3,7-DMU. Conclusions Given the contribution of multiple isoforms to 3-MX and 7-MX formation and the negligible formation of 3,7-DMU in vivo, TB is of little value as a CYP isoform-selective substrate in humans. PMID:10215755

  13. High-flavanol and high-theobromine versus low-flavanol and low-theobromine chocolate to improve uterine artery pulsatility index: a double blind randomized clinical trial.

    Science.gov (United States)

    Bujold, Emmanuel; Leblanc, Vicky; Lavoie-Lebel, Élise; Babar, Asma; Girard, Mario; Poungui, Lionel; Blanchet, Claudine; Marc, Isabelle; Lemieux, Simone; Belkacem, Abdous; Sidi, Elhadji Laouan; Dodin, Sylvie

    2017-09-01

    To evaluate the impact of high-flavanol and high-theobromine (HFHT) chocolate in women at risk of preeclampsia (PE). We conducted a single-center randomized controlled trial including women with singleton pregnancy between 11 and 14 weeks gestation who had bilateral abnormal uterine artery (UtA) waveforms (notching) and elevated pulsatility index (PI). Participants were randomized to either HFHT or low-flavanol and low-theobromine (LFLT) chocolate (30 grams daily for a total of 12 weeks). UtA PI, reported as multiple of medians (MoM) adjusted for gestational age, was assessed at baseline and 12 weeks after randomization. One hundred thirty-one women were randomized with mean gestational age of 12.4 ± 0.6 weeks and a mean UtA PI of 1.39 ± 0.31 MoM. UtA PI adjusted for gestational age significantly decreased from baseline to the second visit (12 weeks later) in the two groups (p chocolate, daily intake of HFHT chocolate was not associated with significant changes of UtA PI. Nevertheless, the improvement observed in both groups suggests that chocolate could improve placental function independently of flavanol and/or theobromine content.

  14. Structure of 2,4-Diaminopyrimidine-Theobromine Alternate Base Pairs

    NARCIS (Netherlands)

    Gengeliczki, Z.; Callahan, M. P.; Kabelac, M.; Rijs, A. M.; de Vries, M. S.

    2011-01-01

    We report the structure of dusters of 2,4-diaminopyrimidine with 3,7-dimethylxanthine (theobromine) in the gas phase determined by IR-UV double resonance spectroscopy in both the near-IR and mid-IR regions in combination with ab initio computations. These clusters represent potential alternate

  15. Method for recovering aroma concentrate from a caffeine- or theobromine-comprising food base material

    NARCIS (Netherlands)

    Kattenberg, H.R.; Willemsen, J.H.A.; Starmans, D.A.J.; Hoving, H.D.; Winters, M.G.M.

    2002-01-01

    Described is a method for recovering aroma concentrate from a caffeine- or theobromine-comprising food base material, such as coffee or tea, and in particular cocoa, at least comprising the steps of: introducing the food base material into an aqueous extractant and incubating the food base material

  16. High performance liquid chromatography determination of theobromine and caffeine in cocoa beans gamma irradiated

    Energy Technology Data Exchange (ETDEWEB)

    Soares, Anderson D.B. [Faculdade de Engenharia de Varginha, MG (Brazil); Mansur Neto, Elias [Minas Gerais Univ., Belo Horizonte, MG (Brazil)

    1997-12-01

    Irradiation is a processing technology that has been shown to be a wholesome process by many scientific studies conducted worldwide during the past 40 years, which has been approved by 37 countries. Irradiated foods have been studied so extensively, that the effects on foods are better understood than any other preservation process, including food freezing and dehydration. Cocoa beans has been commercially irradiated in countries such as Ivory Coast and Argentina. The alkaloids theobromine and caffeine are responsible for the mildly stimulating properties and bitter taste of cocoa and chocolate products. Previously fermented dried and nonfumigated cocoa beans were irradiated at doses of 0, 5.0, 10.0 and 20.0 kGy using Co-60 gamma rays. The samples were analysed for determining theobromine and caffeine contents in the cocoa beans by TIMBIE et al. (1978) high performance liquid chromatography (HPLC) method. Boiling water extracts were cooled, centrifuged and injected into the chromatograph. Theobromine and caffeine were quantitated at 273 nm and showed the tendency of decreasing as the dose of radiation increases. Theobromine and caffeine ranged from 42.3 to 37.1 mg/g and from 7.60 to 6.13 mg/g. respectively from 0 to 20.0 kGy. These results were discussed in relation to the possible acceptance of radiosterization of cocoa beans commercially up to the dose of 20.0 kGy. (author). 10 refs., 1 tab.

  17. Multiple effects of theobromine on fetus development and postnatal status of the immune system.

    Science.gov (United States)

    Chorostowska-Wynimko, J; Skopińska-Rózewska, E; Sommer, E; Rogala, E; Skopiński, P; Wojtasik, E

    2004-01-01

    Caffeine and its active derivative, theobromine, are probably the most frequently ingested pharmacologically active substances. Considering their uninhibited transport via the placental barrier as well as immature enzymatic activities and metabolic pathways in embryos and infants resulting in the longer half-life of methyloxanthines and their accumulation, unrestrained uptake of these substances might result in noticeably more pronounced biological effects during pregnancy and the postnatal period. Our previous studies have shown that methyloxanthines are significant inhibitors of angiogenic growth factors production and angiogenesis itself. We have hypothesized that increased uptake of these substances might affect embryonal angiogenesis and, later in the postnatal period, maturation and functional activity of the offspring's immune system. The study was performed on 2-month-old Balb/c mice fed theobromine 2 or 6 mg/day during pregnancy and lactation. On day 18 of pregnancy the number and weight of embryos were assessed as was their tissue angiogenic activity, using the cutaneous angiogenesis assay. In the group of 4-week-old sucklings, body and spleen were weighed together with the trunk, and tail and limb length were measured. Six weeks after birth the splenocytes' mitogen-induced activity and their ability to induce graft-versus-host reaction as well as the humoral response to SRBC antigen were evaluated. Content of theobromine in the embryos' tissue was estimated by high liquid performance chromatography (HPLC). Theobromine feeding resulted in significant inhibition of embryo growth as assessed by their weight and decreased angiogenic activity of their tissue. The theobromine content in embryo tissue from treated groups was higher than in the controls, and the difference was close to significant. In the postnatal period the discrepancies in the treated 4-week-old group's development were also observed in the significantly shorter limbs in comparison to the

  18. The excretion of theobromine in Thoroughbred racehorses after feeding compounded cubes containing cocoa husk--establishment of a threshold value in horse urine.

    Science.gov (United States)

    Haywood, P E; Teale, P; Moss, M S

    1990-07-01

    Thoroughbred geldings were fed racehorse cubes containing a predetermined concentration of theobromine in the form of cocoa husk. They were offered 7 kg of cubes per day, divided between morning and evening feed, and food consumption was monitored. Urinary concentrations of theobromine were determined following the consumption of cubes containing 11.5, 6.6, 2.0 and 1.2 mg per kg of theobromine, to verify whether or not such concentrations would produce positive urine tests. Pre-dose urine samples were collected to verify the absence of theobromine before each experiment. It became apparent from the results of the first three administrations that the limit of detection of theobromine, using such procedures, would be reached at a feed level of about 1 mg per kg theobromine. Therefore the final administration, using cubes containing 1.2 mg per kg theobromine, was singled out for additional analytical work and quantitative procedures were developed to measure urinary concentrations of theobromine. It was anticipated that the results would form a basis for discussions relating to the establishment of a threshold value for theobromine in horse urine. The Stewards of the Jockey Club subsequently gave notice that they had established a threshold level for theobromine in urine of 2 micrograms/ml.

  19. Salinivibrio costicola GL6, a Novel Isolated Strain for Biotransformation of Caffeine to Theobromine Under Hypersaline Conditions.

    Science.gov (United States)

    Ashengroph, Morahem

    2017-01-01

    The present study has been conducted towards isolation of moderately halophilic bacteria capable of transforming caffeine into theobromine. A total of 45 caffeine-degrading moderate halophiles were enriched from hypersaline lakes and examined for the biotransformation of caffeine to theobromine by thin-layer chromatography (TLC) and high-performance liquid chromatography analyses. Strain GL6, giving the highest yield of theobromine, was isolated from the Hoz Soltan Lake, 20 % w/v salinity, central Iran, and identified as Salinivibrio costicola based on morphological and biochemical features as well as its 16S rRNA gene sequence analysis (GeneBank Accession No. KT378066) and DNA-DNA relatedness. The biotransformation of caffeine with strain GL6 leads to the formation of two metabolites, identified as theobromine and paraxanthine, but the yield of paraxanthine was much lower. Further study on the production of theobromine from caffeine under resting cell experiment was carried out subsequently. The optimal yield of theobromine (56 %) was obtained after a 32-h incubation using 5 mM of caffeine and 15 g l-1 (wet weight) of biomass in 0.1 M saline phosphate buffer (pH 7.0 and 10 % w/v NaCl) under agitation 180 rpm at 30 °C. The biotransformed theobromine was purified by preparative TLC and subjected to FTIR and mass spectroscopy for chemical identification. This is the first evidence for biotransformation of caffeine into theobromine by strains of the genus Salinivibrio.

  20. The effect of theobromine 200 mg/l topical gel exposure duration against surface enamel hardness resistance from 1% citric acid

    Science.gov (United States)

    Herisa, H. M.; Noerdin, A.; Eriwati, Y. K.

    2017-08-01

    Theobromine can be used to prevent the demineralization of enamel and can stimulate the growth of new enamels. This study analyzes the effect of theobromine’s gel duration exposure on enamel hardness resistance from 1% citric acid. Twenty-eight specimens were divided into three experimental groups; were exposed to theobromine gel 200 mg/l for 16, 48, and 96 minutes; and were then immersed in 1% citric acid. The control group was only immersed in 1% citric acid. Results: A Wilcoxon test showed a significant increase and decrease in enamel microhardness after exposure to theobromine gel and citric acid (p theobromine gel and immersion in citric acid (p theobromine gel 200mg/L increased enamel microhardness but did not contribute to the enamel’s hardness resistance after immersion in 1% citric acid. The duration of theobromine gel application affected enamel microhardness and acid resistance.

  1. Theobromine inhibits differentiation of 3T3-L1 cells during the early stage of adipogenesis via AMPK and MAPK signaling pathways.

    Science.gov (United States)

    Jang, Yeon Jeong; Koo, Hyun Jung; Sohn, Eun-Hwa; Kang, Se Chan; Rhee, Dong-Kwon; Pyo, Suhkneung

    2015-07-01

    Obesity is characterized by hypertrophy and/or by the differentiation or adipogenesis of pre-existing adipocytes. In this study, we investigated the inhibitory effects of theobromine, a type of alkaloid in cocoa, on adipocyte differentiation of 3T3-L1 preadipocytes and its mechanisms of action. Theobromine inhibited the accumulation of lipid droplets, the expression of PPARγ and C/EBPα, and the mRNA expression of aP2 and leptin. The inhibition of adipogenic differentiation by theobromine occurred primarily in the early stages of differentiation. In addition, theobromine arrested the cell cycle at the G0/G1 phase and regulated the expressions of CDK2, p27, and p21. Theobromine treatment increased AMPK phosphorylation and knockdown of AMPKα1/α2 prevented the ability of theobromine to inhibit PPARγ expression in the differentiating 3T3-L1 cells. Theobromine reduced the phosphorylation of ERK and JNK. Moreover, the secretion and the mRNA level of TNF-α and IL-6 were inhibited by theobromine treatment. These data suggest that theobromine inhibits adipocyte differentiation during the early stages of adipogenesis by regulating the expression of PPARγ and C/EBPα through the AMPK and ERK/JNK signaling pathways in 3T3-L1 preadipocytes.

  2. Effects of caffeine, theophylline and theobromine on scheduled controlled responding in rats.

    Science.gov (United States)

    Carney, J. M.

    1982-01-01

    1 Rats were trained to respond under a variable interval 30 s (VI 30) schedule of food reinforcement. Caffeine (0.32-32 mg/kg), theophylline (1.0-56 mg/kg) and theobromine (10-320 mg/kg) in general produced dose-related decreases in operant responding. At relatively low doses, caffeine (1.0 mg/kg) and theophylline (3.2 mg/kg) produced slight but nonsignificant increases in VI 30 responding. 3 The rank order of potency for producing decreases in responding was caffeine greater than theophylline greater than theobromine. 4 Daily caffeine injections (32 mg/kg, i.p.) resulted in the development of caffeine tolerance. This tolerance was characterized by a 6 fold shift to the right in the caffeine dose-effect curve. Saline substitution for the 32.0 mg/kg caffeine maintenance dose resulted in a substantial decrease in responding. PMID:7066599

  3. Vibrational spectral investigation on xanthine and its derivatives—theophylline, caffeine and theobromine

    Science.gov (United States)

    Gunasekaran, S.; Sankari, G.; Ponnusamy, S.

    2005-01-01

    A normal coordinate analysis has been carried out on four compounds having a similar ring structure with different side chain substitutions, which are xanthine, caffeine, theophylline, and theobromine. Xanthine is chemically known as 2,6-dihydroxy purine. Caffeine, theophylline and theobromine are methylated xanthines. Considering the methyl groups as point mass, the number of normal modes of vibrations can be distributed as Γ vib=27 A'+12 A″ based on C s point group symmetry associated with the structures. In the present work 15 A' and 12 A″ normal modes are considered. A new set of orthonormal symmetry co-ordinates have been constructed. Wilson's F- G matrix method has been adopted for the normal coordinate analysis. A satisfactory vibrational band assignment has been made by employing the FTIR and FT Raman spectra of the compounds. The potential energy distribution is calculated with the arrived values of the force constants and hence the agreement of the frequency assignment has been checked.

  4. Spectral analysis of naturally occurring methylxanthines (theophylline, theobromine and caffeine) binding with DNA.

    Science.gov (United States)

    Johnson, Irudayam Maria; Prakash, Halan; Prathiba, Jeyaguru; Raghunathan, Raghavachary; Malathi, Raghunathan

    2012-01-01

    Nucleic acids exist in a dynamic equilibrium with a number of molecules that constantly interact with them and regulate the cellular activities. The inherent nature of the structure and conformational integrity of these macromolecules can lead to altered biological activity through proper targeting of nucleic acids binding ligands or drug molecules. We studied the interaction of naturally occurring methylxanthines such as theophylline, theobromine and caffeine with DNA, using UV absorption and Fourier transform infrared (FTIR) spectroscopic methods, and especially monitored their binding affinity in the presence of Mg(2+) and during helix-coil transitions of DNA by temperature (T(m)) or pH melting profiles. The study indicates that all these molecules effectively bind to DNA in a dose dependent manner. The overall binding constants of DNA-theophylline = 3.5×10(3) M(-1), DNA-theobromine = 1.1×10(3) M(-1), and DNA-Caffeine = 3.8×10(3) M(-1). On the other hand T(m)/pH melting profiles showed 24-35% of enhanced binding activity of methylxanthines during helix-coil transitions of DNA rather than to its native double helical structure. The FTIR analysis divulged that theophylline, theobromine and caffeine interact with all the base pairs of DNA (A-T; G-C) and phosphate group through hydrogen bond (H-bond) interaction. In the presence of Mg(2+), methylxanthines altered the structure of DNA from B to A-family. However, the B-family structure of DNA remained unaltered in DNA-methylxanthines complexes or in the absence of Mg(2+). The spectral analyses indicated the order of binding affinity as "caffeine≥theophylline>theobromine" to the native double helical DNA, and "theophylline≥theobromine>caffeine to the denatured form of DNA and in the presence of divalent metal ions.

  5. The relevance of theobromine for the beneficial effects of cocoa consumption.

    OpenAIRE

    Eva eMartínez-Pinilla; Ainhoa eOñatibia-Astibia; Rafael eFranco

    2015-01-01

    Cocoa consumption began in America and in the mid sixteenth Century it quickly spread to Europe. Beyond being considered a pleasant habit due to its rich sweet lingering taste, chocolate was considered a good nutrient and even a medicine. Traditionally, health benefits of cocoa have been related with the high content of antioxidants of Theobroma cocoa beans. However, the direct psychoactive effect due to methylxanthines in cocoa is notable. Theobromine and caffeine, in the proportions found i...

  6. Spectral analysis of naturally occurring methylxanthines (theophylline, theobromine and caffeine binding with DNA.

    Directory of Open Access Journals (Sweden)

    Irudayam Maria Johnson

    Full Text Available Nucleic acids exist in a dynamic equilibrium with a number of molecules that constantly interact with them and regulate the cellular activities. The inherent nature of the structure and conformational integrity of these macromolecules can lead to altered biological activity through proper targeting of nucleic acids binding ligands or drug molecules. We studied the interaction of naturally occurring methylxanthines such as theophylline, theobromine and caffeine with DNA, using UV absorption and Fourier transform infrared (FTIR spectroscopic methods, and especially monitored their binding affinity in the presence of Mg(2+ and during helix-coil transitions of DNA by temperature (T(m or pH melting profiles. The study indicates that all these molecules effectively bind to DNA in a dose dependent manner. The overall binding constants of DNA-theophylline = 3.5×10(3 M(-1, DNA-theobromine = 1.1×10(3 M(-1, and DNA-Caffeine = 3.8×10(3 M(-1. On the other hand T(m/pH melting profiles showed 24-35% of enhanced binding activity of methylxanthines during helix-coil transitions of DNA rather than to its native double helical structure. The FTIR analysis divulged that theophylline, theobromine and caffeine interact with all the base pairs of DNA (A-T; G-C and phosphate group through hydrogen bond (H-bond interaction. In the presence of Mg(2+, methylxanthines altered the structure of DNA from B to A-family. However, the B-family structure of DNA remained unaltered in DNA-methylxanthines complexes or in the absence of Mg(2+. The spectral analyses indicated the order of binding affinity as "caffeine≥theophylline>theobromine" to the native double helical DNA, and "theophylline≥theobromine>caffeine to the denatured form of DNA and in the presence of divalent metal ions.

  7. Method for recovering aroma concentrate from a caffeine- or theobromine-comprising food base material

    OpenAIRE

    Kattenberg, H.R.; Willemsen, J.H.A.; Starmans, D.A.J.; Hoving, H.D.; Winters, M.G.M.

    2002-01-01

    Described is a method for recovering aroma concentrate from a caffeine- or theobromine-comprising food base material, such as coffee or tea, and in particular cocoa, at least comprising the steps of: introducing the food base material into an aqueous extractant and incubating the food base material at a suitable temperature and for a suitable period, substantial extraction of the aromas taking place to form an aqueous food extract, pervaporating the food extract using a hydrophobic pervaporat...

  8. Nilai Cerna dan Biodegradasi Theobromin Pod Kakao dengan Perlakuan Fermentasi Menggunakan Inokulum Multi Mikrobia

    OpenAIRE

    Wulandari, Suci; Agus, Ali; Soejono, Mohamad; Cahyanto, Muhammad Nur

    2014-01-01

    In the recent years it has been reported that there is a multimicrobe culture which is used for fermentation of feed ingredients and complete feeds. This multi microbial culture is introduced by the name of SBP® (Saus Burger Feed). This research study aims to understand the influence of fermented cocoa pod using multi microbe which is contained in the SBP to digest and biodegrade theobromine in the cocoo pod. Cocoa pod was fermented under anaerobic conditions at room temperature. Inoculums le...

  9. Nilai Cerna dan Biodegradasi Theobromin Pod Kakao dengan Perlakuan Fermentasi Menggunakan Inokulum Multi Mikrobia

    OpenAIRE

    Suci Wulandari; Ali Agus; Mohamad Soejono; Muhammad Nur Cahyanto

    2014-01-01

    In the recent years it has been reported that there is a multimicrobe culture which is used for fermentation of feed ingredients and complete feeds. This multi microbial culture is introduced by the name of SBP® (Saus Burger Feed). This research study aims to understand the influence of fermented cocoa pod using multi microbe which is contained in the SBP to digest and biodegrade theobromine in the cocoo pod. Cocoa pod was fermented under anaerobic conditions at room temperature. Inoculums...

  10. Spectral Analysis of Naturally Occurring Methylxanthines (Theophylline, Theobromine and Caffeine) Binding with DNA

    OpenAIRE

    Irudayam Maria Johnson; Halan Prakash; Jeyaguru Prathiba; Raghavachary Raghunathan; Raghunathan Malathi

    2012-01-01

    Nucleic acids exist in a dynamic equilibrium with a number of molecules that constantly interact with them and regulate the cellular activities. The inherent nature of the structure and conformational integrity of these macromolecules can lead to altered biological activity through proper targeting of nucleic acids binding ligands or drug molecules. We studied the interaction of naturally occurring methylxanthines such as theophylline, theobromine and caffeine with DNA, using UV absorption an...

  11. Phenolic and Theobromine Contents of Commercial Dark, Milk and White Chocolates on the Malaysian Market

    Directory of Open Access Journals (Sweden)

    Cheng Chia Meng

    2009-01-01

    Full Text Available 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.

  12. Phenolic and theobromine contents of commercial dark, milk and white chocolates on the Malaysian market.

    Science.gov (United States)

    Meng, Cheng Chia; Jalil, Abbe Maleyki Mhd; Ismail, Amin

    2009-01-05

    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.

  13. Effects of caffeine and its reactive metabolites theophylline and theobromine on the differentiating testis.

    Science.gov (United States)

    Pollard, I; Locquet, O; Solvar, A; Magre, S

    2001-01-01

    A previous study in the rat (Pollard et al. 1990) established that caffeine, when administered during pregnancy, significantly inhibited the differentiation of the seminiferous cords and subsequent Leydig cell development in the interstitium. However, that study could not distinguish between the direct effects of caffeine and/or the intermediary secondary toxic effects of metabolites such as theophylline and theobromine. Because the fetus lacks the appropriate enzyme systems, clearance of toxic substances takes place via the placenta and maternal liver. Thus, a suitable in vitro system can effectively differentiate between primary and secondary drug effects. In the present study, 13-day-old fetal testis, at the stage of incipient differentiation, were cultured for 4 days in vitro in the presence of graded doses of caffeine, theophylline or theobromine. It was found that explants exposed to caffeine or theobromine differentiated normally, developing seminiferous cords made up of Sertoli and germ cells, soon followed by the differentiation of functionally active Leydig cells appearing in the newly formed interstitium. However, explants exposed to theophylline failed to develop seminiferous cords and, as a consequence, Leydig cells. In conclusion, insights obtained from different experimental methods, such as organ culture or whole organism studies, are not always identical. It may be prudent, therefore, to take into account that certain experimental techniques, despite providing valuable information, may require confirmation by other test methods in order to obtain an in-depth understanding of mechanisms of action involved.

  14. Flavan-3-ols, theobromine, and the effects of cocoa and chocolate on cardiometabolic risk factors.

    Science.gov (United States)

    Berends, Lindsey M; van der Velpen, Vera; Cassidy, Aedin

    2015-02-01

    Although there is growing interest surrounding the potential health benefits of cocoa and chocolate, the relative contribution of bioactive constituents for these effects remains unclear. This review summarizes the recent research on the cardiometabolic effects of cocoa and chocolate with a focus on two key constituents: flavan-3-ols and theobromine. Recent meta-analyses suggest beneficial cardiometabolic effects of chocolate following short-term intake, including improvements in flow-mediated dilatation, blood pressure, lipoprotein levels and biomarkers of insulin resistance. Flavan-3-ols may play a role, but it is currently unclear which specific compounds or metabolites are key. Theobromine has also been shown to improve lipoprotein levels in trials, although these findings need verification at habitual intake levels. Longer term dose-response randomized controlled trials are required to determine the sustainability of the short-term effects and the optimal dose. Quantifying levels of bioactives in intervention products and their metabolites in biological samples will facilitate the assessment of their relative impact and the underlying mechanisms of action. Promising data support the beneficial cardiometabolic effects of cocoa and chocolate intake, with significant interest in the flavan-3-ol and theobromine content. Validated biomarkers of intake together with more relevant mechanistic insights from experimental models using physiologically relevant concentrations and metabolites will continue to inform this research field.

  15. Effect of theobromine-containing toothpaste on dentin tubule occlusion in situ.

    Science.gov (United States)

    Amaechi, Bennett T; Mathews, Sapna M; Mensinkai, Poornima K

    2015-01-01

    Dentin hypersensitivity (DH) is treated by either occlusion of dentin tubules or nerve desensitization. This in situ study compared dentin tubules occlusion by theobromine-containing dentifrices with (Theodent-classic-F®, TCF) and without (Theodent-classic®, TC) fluoride with 1,500 ppm fluoride toothpaste, Colgate®-Regular (Fluoride) and Novamin®-containing toothpaste, Sensodyne®-5000-Nupro (Novamin®). Each subject wore four intraoral appliances bearing dentin blocks while using one of four test dentifrices (n = 20/dentifrice) twice daily for 7 days. The four appliances were removed successively after 1, 2, 3, and 7 days. Treated blocks and their control (untreated) blocks were examined with scanning electron microscopy (SEM). Effects were compared statistically (ANOVA/Tukey's) based on percentage of surface area covered by deposited precipitate layer (%DPL) and percentage of fully open (%FOT), partially occluded (%POT), and completely occluded (%COT) tubules in each block calculated relative to the number of tubules in their control blocks. SEM observation indicated an increased %COT and %DPL over time. After 1 and 2 days, %COT was comparable with TC and TCF, and significantly (p Theobromine-containing toothpastes with and without fluoride have equal potential in occluding dentin tubules within a shorter time period than Novamin®-containing toothpaste; however, the three demonstrated equal potential after 1 week, but not the fluoride toothpaste. Theobromine-containing toothpaste promoted dentin tubule occlusion thus shows potential to relief DH.

  16. Effects on Peripheral and Central Blood Pressure of Cocoa With Natural or High-Dose Theobromine A Randomized, Double-Blind Crossover Trial

    NARCIS (Netherlands)

    van den Bogaard, Bas; Draijer, Richard; Westerhof, Berend E.; van den Meiracker, Anton H.; van Montfrans, Gert A.; van den Born, Bert-Jan H.

    2010-01-01

    Flavanol-rich cocoa products have been reported to lower blood pressure. It has been suggested that theobromine is partially responsible for this effect. We tested whether consumption of flavanol-rich cocoa drinks with natural or added theobromine could lower peripheral and central blood pressure.

  17. Theobromine does not affect postprandial lipid metabolism and duodenal gene expression, but has unfavorable effects on postprandial glucose and insulin responses in humans

    NARCIS (Netherlands)

    Smolders, Lotte; Mensink, Ronald P.; Boekschoten, Mark V.; Ridder, de Rogier J.J.; Plat, Jogchum

    2017-01-01

    Background & aims: Chocolate consumption is associated with a decreased risk for CVD. Theobromine, a compound in cocoa, may explain these effects as it favorably affected fasting serum lipids. However, long-term effects of theobromine on postprandial metabolism as well as underlying

  18. Effects on peripheral and central blood pressure of cocoa with natural or high-dose theobromine: A randomized, double-blind crossover trial

    NARCIS (Netherlands)

    B. van den Bogaard (Bas); R. Draijer (Richard); B.E. Westerhof (Berend); A.H. van den Meiracker (Anton); G.A. van Montfrans (Gert); B.J.H. van den Born

    2010-01-01

    textabstractFlavanol-rich cocoa products have been reported to lower blood pressure. It has been suggested that theobromine is partially responsible for this effect. We tested whether consumption of flavanol-rich cocoa drinks with natural or added theobromine could lower peripheral and central blood

  19. Methylxanthines accumulation in Ilex species - caffeine and theobromine in erva-mate (Ilex paraguariensis and other Ilex species

    Directory of Open Access Journals (Sweden)

    Reginatto F.H.

    1999-01-01

    Full Text Available Ilex paraguariensis St.Hil. is an important crop used commonly at the meridional South America as a source of a stimulant beverage, called maté, prepared by infusion of its processed leaves and twigs. We describe herein the methylxanthines analyses in the leaves of I. paraguariensis and other Ilex species reported as maté adulterants. The methylxanthines content determined by HPLC were 0.65% for caffeine and 0.12% for theobromine from I. paraguariensis var. paraguariensis, and 0.003% for caffeine and 0.22% (estimated for theobromine from I. paraguariensis var. vestita. It was not detected any methylxanthines from I. brevicuspis, I. dumosa and I. microdonta. Considering the results, simultaneous caffeine and theobromine accumulation seems hitherto to be a particular characteristic of I. paraguariensis. In addition to taxonomical significance, these data suggest that it should be possible to trace adulterations of the genuine I. paraguariensis using the methodology described herein.

  20. Spectral Analysis of Naturally Occurring Methylxanthines (Theophylline, Theobromine and Caffeine) Binding with DNA

    Science.gov (United States)

    Johnson, Irudayam Maria; Prakash, Halan; Prathiba, Jeyaguru; Raghunathan, Raghavachary; Malathi, Raghunathan

    2012-01-01

    Nucleic acids exist in a dynamic equilibrium with a number of molecules that constantly interact with them and regulate the cellular activities. The inherent nature of the structure and conformational integrity of these macromolecules can lead to altered biological activity through proper targeting of nucleic acids binding ligands or drug molecules. We studied the interaction of naturally occurring methylxanthines such as theophylline, theobromine and caffeine with DNA, using UV absorption and Fourier transform infrared (FTIR) spectroscopic methods, and especially monitored their binding affinity in the presence of Mg2+ and during helix-coil transitions of DNA by temperature (Tm) or pH melting profiles. The study indicates that all these molecules effectively bind to DNA in a dose dependent manner. The overall binding constants of DNA-theophylline = 3.5×103 M−1, DNA-theobromine = 1.1×103 M−1, and DNA-Caffeine = 3.8×103 M−1. On the other hand Tm/pH melting profiles showed 24–35% of enhanced binding activity of methylxanthines during helix-coil transitions of DNA rather than to its native double helical structure. The FTIR analysis divulged that theophylline, theobromine and caffeine interact with all the base pairs of DNA (A-T; G-C) and phosphate group through hydrogen bond (H-bond) interaction. In the presence of Mg2+, methylxanthines altered the structure of DNA from B to A-family. However, the B-family structure of DNA remained unaltered in DNA-methylxanthines complexes or in the absence of Mg2+. The spectral analyses indicated the order of binding affinity as “caffeine≥theophylline>theobromine” to the native double helical DNA, and “theophylline≥theobromine>caffeine to the denatured form of DNA and in the presence of divalent metal ions. PMID:23236361

  1. Determination of caffeine, theophylline and theobromine in serum and saliva using high-performance liquid chromatography.

    Science.gov (United States)

    Scott, N R; Chakraborty, J; Marks, V

    1984-03-01

    A method is described for the measurement of theobromine, theophylline and caffeine in serum and saliva by high-performance liquid chromatography (HPLC). A chloroform/isopropanol extract (85:15, v/v) is evaporated to dryness and chromatographed on a 100 X 4.5 mm id Hypersil octadecylsilane column with UV detection at 280 nm. Theobromine, theophylline, caffeine and the internal standard proxyphylline are satisfactorily resolved with an elution system of acetonitrile/tetrahydrofuran/50 mM acetate buffer, pH 4.0, (4:1:95, v/v). No interference is observed from the presence of xanthine metabolites or any of a number of common drugs examined. A good correlation was observed between the concentrations of caffeine in serum and in saliva suggesting that salivary measurements may be useful for the study of caffeine pharmacokinetics in man. Caffeine levels determined by the HPLC procedure described here agreed well with those obtained by a radioimmunoassay method. The method is also suitable for determining the xanthine content of beverages by direct injection of diluted samples.

  2. Simultaneous HPLC determination of caffeine, theobromine, and theophylline in food, drinks, and herbal products.

    Science.gov (United States)

    Srdjenovic, Branislava; Djordjevic-Milic, Vukosava; Grujic, Nevena; Injac, Rade; Lepojevic, Zika

    2008-02-01

    A rapid and selective high-performance liquid chromatographic (HPLC) method is developed for the separation and determination of caffeine, theobromine, and theophylline. The chromatography is performed on a Zorbax Eclipse XDB-C8 column (4.6x150 mm i.d., 5-microm particle size) at 25 degrees C, with a mobile phase of water-THF (0.1% THF in water, pH 8)-acetonitrile (90:10, v/v). The flow rate is 0.8 mL/min, and detection is by UV at 273 nm. This method permits the simultaneous determination of caffeine, theobromine, and theophylline in food, drinks, and herbal products with detection limits of 0.07-0.2 mg/L and recoveries of 100.20-100.42%. Correlation coefficients, for the calibration curves in the linear range of 0.2-100 mg/L, are greater than 0.9999 for all compounds. The within- and between-day precision is determined for both retention times and peak area. The data suggests that the proposed HPLC method can be used for routine quality control of food, drinks, and herbal products.

  3. Simultaneous determination of caffeine, theobromine, and theophylline by high-performance liquid chromatography.

    Science.gov (United States)

    Bispo, Marcia S; Veloso, Márcia Cristina C; Pinheiro, Heloísa Lúcia C; De Oliveira, Rodolfo F S; Reis, José Oscar N; De Andrade, Jailson B

    2002-01-01

    This work relates the development of an analytical methodology to simultaneously determine three methylxanthines (caffeine, theobromine, and theophylline) in beverages and urine samples based on reversed-phase high-performance liquid chromatography. Separation is made with a Bondesil C18 column using methanol-water-acetic acid or ethanol-water-acetic acid (20:75:5, v/v/v) as the mobile phase at 0.7 mL/min. Identification is made by absorbance detection at 273 nm. Under optimized conditions, the detection limit of the HPLC method is 0.1 pg/mL for all three methylxanthines. This method is applied to urine and to 25 different beverage samples, which included coffee, tea, chocolate, and coconut water. The concentration ranges determined in the beverages and urine are: theobromine; < 0.1 pg/mL to 47 microg/mL and < 0.1 pg/mL to 66.3 microg/mL for theophylline. The method proposed in this study is rapid and suitable for the simultaneous quantitation of methylxanthines in beverages and human urine samples and requires no extraction step or derivatization.

  4. Vibrational spectral investigation on xanthine and its derivatives--theophylline, caffeine and theobromine.

    Science.gov (United States)

    Gunasekaran, S; Sankari, G; Ponnusamy, S

    2005-01-01

    A normal coordinate analysis has been carried out on four compounds having a similar ring structure with different side chain substitutions, which are xanthine, caffeine, theophylline, and theobromine. Xanthine is chemically known as 2,6-dihydroxy purine. Caffeine, theophylline and theobromine are methylated xanthines. Considering the methyl groups as point mass, the number of normal modes of vibrations can be distributed as Gamma(vib) = 27 A' + 12 A" based on C(s) point group symmetry associated with the structures. In the present work 15 A' and 12 A'' normal modes are considered. A new set of orthonormal symmetry co-ordinates have been constructed. Wilson's F-G matrix method has been adopted for the normal coordinate analysis. A satisfactory vibrational band assignment has been made by employing the FTIR and FT Raman spectra of the compounds. The potential energy distribution is calculated with the arrived values of the force constants and hence the agreement of the frequency assignment has been checked.

  5. Simultaneous determination of caffeine, theophylline and theobromine in food samples by a kinetic spectrophotometric method.

    Science.gov (United States)

    Xia, Zhenzhen; Ni, Yongnian; Kokot, Serge

    2013-12-15

    A novel kinetic spectrophotometric method was developed for the simultaneous determination of caffeine, theobromine and theophylline in food samples. This method was based on the different kinetic characteristics between the reactions of analytes with cerium sulphate in sulphuric acid and the associated change in absorbance at 320 nm. Experimental conditions, the effects of sulphuric acid, cerium sulphate and temperature, were optimised. Linear ranges (0.4-8.4 μg mL(-1)) for all three analytes were established, and the limits of detection were: 0.30 μg mL(-1) (caffeine), 0.33 μg mL(-1) (theobromine) and 0.16 μg mL(-1) (theophylline). The recorded data were processed by partial least squares and artificial neural network, and the developed mathematical models were then used for prediction. The proposed, novel method was applied to determine the analytes in commercial food samples, and there were no significant differences between the results from the proposed method and those obtained by high-performance liquid chromatography. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. The effects of brushing on human enamel surface roughness after NaF gel and theobromine gel exposure

    Science.gov (United States)

    Mahardhika, A.; Noerdin, A.; Eriwati, Y. K.

    2017-08-01

    This study aimed to determine the effects of brushing on human enamel surface roughness after different exposure times of 200 mg/L theobromine gel (8, 16, and 32 minutes) and 2% NaF gel (16 minutes). Twenty-four human upper premolars were used and divided into four groups. Group 1 was exposed to 2% NaF gel for 16 minutes. In contrast, groups 2, 3, and 4 were exposed to 200 mg/L theobromine gel for 8 minutes, 16 minutes, and 32 minutes, and each group was then brushed for 9 minutes and 20 seconds. After the treatment, samples were tested using a surface roughness tester (Mitutoyo SJ 301, Japan). The Wilcoxon test showed significant changes (p < 0.05) in roughness values after exposure to the theobromine gel or NaF gel and after brushing for 9 minutes and 20 seconds. It can be concluded that exposure to 200 mg/L theobromine gel or 2% NaF gel can soften the enamel surface and then increase roughness after brushing.

  7. Catecholaminergic and cholinergic systems of mouse brain are modulated by LMN diet, rich in theobromine, polyphenols and polyunsaturated fatty acids.

    Science.gov (United States)

    Fernández-Fernández, Laura; Esteban, Gerard; Giralt, Mercedes; Valente, Tony; Bolea, Irene; Solé, Montse; Sun, Ping; Benítez, Susana; Morelló, José Ramón; Reguant, Jordi; Ramírez, Bartolomé; Hidalgo, Juan; Unzeta, Mercedes

    2015-04-01

    The possible modulatory effect of the functional LMN diet, rich in theobromine, polyphenols and polyunsaturated fatty acids, on the catecholaminergic and cholinergic neurotransmission, affecting cognition decline during aging has been studied. 129S1/SvlmJ mice were fed for 10, 20, 30 and 40 days with either LMN or control diets. The enzymes involved in catecholaminergic and cholinergic metabolism were determined by both immunohistological and western blot analyses. Noradrenalin, dopamine and other metabolites were quantified by HPLC analysis. Theobromine, present in cocoa, the main LMN diet component, was analysed in parallel using SH-SY5Y and PC12 cell lines. An enhanced modulatory effect on both cholinergic and catecholaminergic transmissions was observed on 20 day fed mice. Similar effect was observed with theobromine, besides its antioxidant capacity inducing SOD-1 and GPx expression. The enhancing effect of the LMN diet and theobromine on the levels of acetylcholine-related enzymes, dopamine and specially noradrenalin confirms the beneficial role of this diet on the "cognitive reserve" and hence a possible reducing effect on cognitive decline underlying aging and Alzheimer's disease.

  8. Determination of the Antiproliferative Activity of New Theobromine Derivatives and Evaluation of Their In Vitro Hepatotoxic Effects.

    Science.gov (United States)

    Georgieva, Maya; Kondeva-Burdina, Magdalena; Mitkov, Javor; Tzankova, Virginia; Momekov, Georgi; Zlatkov, Alexander

    2016-01-01

    A new series of N-substituted 1-benzyltheobromine-8-thioacetamides were designed and synthesized. Their anti-proliferative activity against human chronic myelocytic leukemia cell K562, human T-cell leukemia cell SKW-3 and human acute myeloid leukemia HL-60 was evaluated. For the tested compounds a concentrationdependent cytotoxic activity was observed, with 7g outlined as the most active compound within the series. The targed compounds were obtained in yields of 56 to 85% and their structures were elucidated by FTIR, (1)H NMR, (13)C NMR and microanalyses. The compounds purity was proven by elemental analysis and spectral data. In general, the compounds showed low hepatotoxicity on sub-cellular and cellular level. On isolated rat microsomes only 7d showed toxic effect while theobromine, 1-benzyl-theobromine-thioacetic acid (BTTA) and the other new theobromine derivatives were devoid of toxicity. In isolated rat hepatocytes, when compared to theobromine and BTTA, 7f showed lower cytotoxic effects, and 7d exerted higher cytotoxicity. The results indicate 7g as a promising structure for the design of future compounds with low hepatotoxicity and good antiproliferative activity.

  9. Determination of caffeine, theobromine, and theophylline in standard reference material 2384, baking chocolate, using reversed-phase liquid chromatography.

    Science.gov (United States)

    Thomas, Jeanice Brown; Yen, James H; Schantz, Michele M; Porter, Barbara J; Sharpless, Katherine E

    2004-06-02

    A rapid and selective isocratic reversed-phase liquid chromatographic method has been developed at the National Institute of Standards and Technology to simultaneously measure caffeine, theobromine, and theophylline in a food-matrix standard reference material (SRM) 2384, Baking Chocolate. The method uses isocratic elution with a mobile phase composition (volume fractions) of 10% acetronitrile/90% water (pH adjusted to 2.5 using acetic acid) at a flow rate of 1.5 mL/min with ultraviolet absorbance detection (274 nm). Total elution time for these analytes is less than 15 min. Concentration levels of caffeine, theobromine, and theophylline were measured in single 1-g samples taken from each of eight bars of chocolate over an eight-day period. Samples were defatted with hexane, and beta-hydroxyethyltheophylline was added as the internal standard. The repeatability for the caffeine, theobromine, and theophylline measurements was 5.1, 2.3, and 1.9%, respectively. The limit of quantitation for all analytes was theobromine, and theophylline in SRM 2384.

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

    Science.gov (United States)

    Hasegawa, Takashi; Takahashi, Kazunaga; Saijo, Masaaki; Ishii, Toshiyasu; Nagata, Tomoko

    2009-12-01

    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.

  11. Inhibitory effect of theobromine on induction of angiogenesis and VEGF mRNA expression in v-raf transfectants of human urothelial cells HCV-29.

    Science.gov (United States)

    Skopinska-Rózewska, E; Janik, P; Przybyszewska, M; Sommer, E; Bialas-Chromiec, B

    1998-12-01

    Neovascularisation plays a crucial role in solid tumor growth and metastasis formation. Our previous studies showed that theophylline and theobromine suppressed cutaneous neovascular reaction induced in mice by human blood leukocytes, and lung as well as ovarian cancer cells. Here, we investigated the in vivo effect of theobromine on angiogenic activity of human urothelial cell line HCV-29, v-raf transfected (mouse cutaneous assay), and the in vitro effect of this drug on VEGF, tPA, uPA and PAI mRNA expression in these cells (RT-PCR method). Theobromine suppressed angiogenesis induced in mice by HCV-29-v-raf cells, inhibited VEGF mRNA expression, and had no effect on transcription of uPA and tPA in these cells. HCV-29-v-raf transfectants do not display transcripts of PAI, in the presence or the absence of theobromine.

  12. Effects on peripheral and central blood pressure of cocoa with natural or high-dose theobromine: a randomized, double-blind crossover trial.

    Science.gov (United States)

    van den Bogaard, Bas; Draijer, Richard; Westerhof, Berend E; van den Meiracker, Anton H; van Montfrans, Gert A; van den Born, Bert-Jan H

    2010-11-01

    Flavanol-rich cocoa products have been reported to lower blood pressure. It has been suggested that theobromine is partially responsible for this effect. We tested whether consumption of flavanol-rich cocoa drinks with natural or added theobromine could lower peripheral and central blood pressure. In a double-blind, placebo-controlled 3-period crossover trial we assigned 42 healthy individuals (age 62±4.5 years; 32 men) with office blood pressure of 130 to 159 mm Hg/85 to 99 mm Hg and low added cardiovascular risk to a random treatment sequence of dairy drinks containing placebo, flavanol-rich cocoa with natural dose consisting of 106 mg of theobromine, or theobromine-enriched flavanol-rich cocoa with 979 mg of theobromine. Treatment duration was 3 weeks with a 2-week washout. The primary outcome was the difference in 24-hour ambulatory systolic blood pressure between placebo and active treatment after 3 weeks. The difference in central systolic blood pressure between placebo and active treatment was a secondary outcome. Treatment with theobromine-enriched cocoa resulted in a mean±SE of 3.2±1.1 mm Hg higher 24-hour ambulatory systolic blood pressure compared with placebo (Ptheobromine-enriched cocoa, laboratory peripheral systolic blood pressure was not different from placebo, whereas central systolic blood pressure was 4.3±1.4 mm Hg lower (P=0.001). Natural dose theobromine cocoa did not significantly change either 24-hour ambulatory or central systolic blood pressure compared with placebo. In conclusion, theobromine-enriched cocoa significantly increased 24-hour ambulatory systolic blood pressure while lowering central systolic blood pressure.

  13. Assessment of flavanol stereoisomers and caffeine and theobromine content in commercial chocolates.

    Science.gov (United States)

    Alañón, M E; Castle, S M; Siswanto, P J; Cifuentes-Gómez, T; Spencer, J P E

    2016-10-01

    Assessment of the flavanol composition of 41 commercial chocolates was by HPLC-DAD. Among individual flavonols ranged from 0.095 to 3.264mgg(-1), epicatechin was the predominant flavanol accounting for 32.9%. Contrary to catechin, epicatechin was a reliable predictive value of the polyphenol content. Conversely the percentage of theobromine used as a proxy measure for nonfat cocoa solids (NFCS) was not a good predictor of epicatechin or flavanol content. In a further chiral analysis, the naturally occurring forms of cocoa flavanols, (-)-epicatechin and (+)-catechin, was determined joint the occurrence of (+)-epicatechin and (-)-catechin due to the epimerization reactions produced in chocolate manufacture. (-)-Epicatechin, the most bioactive compound and predominant form accounted of 93%. However, no positive correlation was found with% cocoa solids, the most significant quality parameter. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Binding of caffeine, theophylline, and theobromine with human serum albumin: A spectroscopic study

    Science.gov (United States)

    Zhang, Hong-Mei; Chen, Ting-Ting; Zhou, Qiu-Hua; Wang, Yan-Qing

    2009-12-01

    The interaction between three purine alkaloids (caffeine, theophylline, and theobromine) and human serum albumin (HSA) was investigated using UV/vis absorption, circular dichroism (CD), fluorescence, synchronous fluorescence, and three-dimensional fluorescence spectra techniques. The results revealed that three alkaloids caused the fluorescence quenching of HSA by the formation of alkaloid-HSA complex. The binding site number n and apparent binding constant KA, corresponding thermodynamic parameters the free energy change (Δ G), enthalpy change (Δ H), and entropy change (Δ S) at different temperatures were calculated. The hydrophobic interaction plays a major role in stabilizing the complex. The distance r between donor (HSA) and acceptor (alkaloids) was obtained according to fluorescence resonance energy transfer. The effect of alkaloids on the conformation of HSA was analyzed using circular dichroism (CD), UV/vis absorption, synchronous fluorescence and three-dimensional fluorescence spectra techniques.

  15. The effect of time in the exposure of theobromine gel to enamel and surface hardness after demineralization with 1% citric acid

    Science.gov (United States)

    Irawan, M. I. P.; Noerdin, A.; Eriwati, Y. K.

    2017-08-01

    Theobromine is one of the alkaloid compounds that can be found in cacao (Theobroma cacao). It is said that theobromine can prevent enamel demineralization. The aim of this research was to evaluate the effect of different exposure times to 200 mg/L theobromine gel on enamel microhardness after demineralization in 1% citric acid. Twenty-eight specimens of human premolar teeth were divided into four groups and were immersed in 1% citric acid (pH 4) for 2.5 minutes. Then 200 mg/L theobromine gel was exposed to the specimens for 16 minutes (n = 7), 48 minutes (n = 7), and 96 minutes (n = 7). Enamel microhardness (KHN) values were tested using the Knoop Microhardness Tester (Shimadzu, Japan) using a 50-gram load for 5 seconds. A statistical test was performed using the Friedman test, Wilcoxon test, Kruskal-Wallis test, and Mann-Whitney test. The results showed a significant decrease, of microhardness values after demineralization with 1% citric acid. There was also a significant increase in hardness (p<0.05) after exposure of the demineralized specimens to theobromine gel for 16 minutes (32.3%), 48 minutes (39.8%), and 96 minutes (43.7%). It can be concluded that exposure to 200 mg/L theobromine gel for 16, 48, and 96 minutes increased enamel microhardness.

  16. Improved high-performance liquid chromatography method to determine theobromine and caffeine in cocoa and cocoa products.

    Science.gov (United States)

    Pura Naik, J

    2001-08-01

    At present, the commonly used HPLC method for the analysis of caffeine and theobromine contents in aqueous cocoa extracts employs direct application of the extracts on the column. This practice gradually reduces the efficiency of the column and shortens its life. Also, this method gives inflated values due to interfering substances and difficulty in achieving baseline resolution. In the improved method, the interfering cocoa pigments are effectively removed by passing the aqueous extract through a Sep-pak C(18) cartridge. Subsequent injection on a C(18) reverse-phase column employing acetonitrile and water (20:80) as the mobile phase reduces the analysis time without affecting either resolution of the peak or the accuracy of caffeine and theobromine determination or achieving baseline resolution. Therefore, this method is ideally suited for rapid routine analysis of cocoa and its products.

  17. Theoretical study on the N-demethylation mechanism of theobromine catalyzed by P450 isoenzyme 1A2.

    Science.gov (United States)

    Tao, Jing; Kang, Yuan; Xue, Zhiyu; Wang, Yongting; Zhang, Yan; Chen, Qiu; Chen, Zeqin; Xue, Ying

    2015-09-01

    Theobromine, a widely consumed pharmacological active substance, can cause undesirable muscle stiffness, nausea and anorexia in high doses ingestion. The main N-demethylation metabolic mechanism of theobromine catalyzed by P450 isoenzyme 1A2 (CYP1A2) has been explored in this work using the unrestricted hybrid density functional method UB3LYP in conjunction with the LACVP(Fe)/6-31G (H, C, N, O, S, Cl) basis set. Single-point calculations including empirical dispersion corrections were carried out at the higher 6-311++G** basis set. Two N-demethylation pathways were characterized, i.e., 3-N and 7-N demethylations, which involve the initial N-methyl hydroxylation to form carbinolamines and the subsequent carbinolamines decomposition to yield monomethylxanthines and formaldehydes. Our results have shown that the rate-limiting N-methyl hydroxylation occurs via a hydrogen atom transfer (HAT) mechanism, which proceeds in a spin-selective mechanism (SSM) in the gas phase. The carbinolamines generated are prone to decomposition via the contiguous heteroatom-assisted proton-transfer. Strikingly, 3-N demethylation is more favorable than 7-N demethylation due to its lower free energy barrier and 7-methylxanthine therefore is the optimum product reported for the demethylation of theobromine catalyzed by CYP1A2, which are in good agreement with the experimental observation. This work has first revealed the detail N-demethylation mechanisms of theobromine at the theoretical level. It can offer more significant information for the metabolism of purine alkaloid. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Effect of Theobromine Consumption on Serum Lipoprotein Profiles in Apparently Healthy Humans with Low HDL-Cholesterol Concentrations.

    Science.gov (United States)

    Jacobs, Doris M; Smolders, Lotte; Lin, Yuguang; de Roo, Niels; Trautwein, Elke A; van Duynhoven, John; Mensink, Ronald P; Plat, Jogchum; Mihaleva, Velitchka V

    2017-01-01

    Scope: Theobromine is a major active compound in cocoa with allegedly beneficial effect on high-density-lipoprotein-cholesterol (HDL-CH). We have investigated the effect of theobromine (TB) consumption on the concentrations of triglyceride (TG) and cholesterol (CH) in various lipoprotein (LP) subclasses. Methods: In a randomized, double-blind, placebo-controlled, cross-over study, 44 apparently healthy women and men (age: 60 ± 6 years, BMI: 29 ± 3 kg/m2) with low baseline HDL-CH concentrations consumed a drink supplemented with 500 mg/d theobromine for 4 weeks. TG and CH concentrations in 15 LP subclasses were predicted from diffusion-edited 1H NMR spectra of fasting serum. Results: The LP phenotype of the subjects was characterized by low CH concentrations in the large HDL particles and high TG concentrations in large VLDL and chylomicron (CM) particles, which clearly differed from a LP phenotype of subjects with normal HDL-CH. TB only reduced CH concentrations in the LDL particles by 3.64 and 6.79%, but had no effect on TG and CH in any of the HDL, VLDL and CM subclasses. Conclusion: TB was not effective on HDL-CH in subjects with a LP phenotype characterized by low HDL-CH and high TG in VLDL.

  19. Effect of Theobromine Consumption on Serum Lipoprotein Profiles in Apparently Healthy Humans with Low HDL-Cholesterol Concentrations

    Directory of Open Access Journals (Sweden)

    Doris M. Jacobs

    2017-08-01

    Full Text Available Scope: Theobromine is a major active compound in cocoa with allegedly beneficial effect on high-density-lipoprotein-cholesterol (HDL-CH. We have investigated the effect of theobromine (TB consumption on the concentrations of triglyceride (TG and cholesterol (CH in various lipoprotein (LP subclasses.Methods: In a randomized, double-blind, placebo-controlled, cross-over study, 44 apparently healthy women and men (age: 60 ± 6 years, BMI: 29 ± 3 kg/m2 with low baseline HDL-CH concentrations consumed a drink supplemented with 500 mg/d theobromine for 4 weeks. TG and CH concentrations in 15 LP subclasses were predicted from diffusion-edited 1H NMR spectra of fasting serum.Results: The LP phenotype of the subjects was characterized by low CH concentrations in the large HDL particles and high TG concentrations in large VLDL and chylomicron (CM particles, which clearly differed from a LP phenotype of subjects with normal HDL-CH. TB only reduced CH concentrations in the LDL particles by 3.64 and 6.79%, but had no effect on TG and CH in any of the HDL, VLDL and CM subclasses.Conclusion: TB was not effective on HDL-CH in subjects with a LP phenotype characterized by low HDL-CH and high TG in VLDL.

  20. Control of methylxanthines in the competition horse: pharmacokinetic/pharmacodynamic studies on caffeine, theobromine and theophylline for the assessment of irrelevant concentrations.

    Science.gov (United States)

    Machnik, Marc; Kaiser, Simone; Koppe, Sophie; Kietzmann, Manfred; Schenk, Ina; Düe, Michael; Thevis, Mario; Schänzer, Wilhelm; Toutain, Pierre-Louis

    2017-09-01

    Methylxanthines positives in competition samples have challenged doping control laboratories and racing jurisdictions since methylxanthines are naturally occurring prohibited substances and often constituents of feed. For theobromine, an international threshold (renamed in International Residue Limit, IRL) of 2 µg/mL in urine has been established. On the basis of the data presented herein, a threshold or rather an IRL for theobromine in plasma of 0.3 µg/mL was proposed and was thereupon approved by the International Federation of Horseracing Authorities (IFHA). Official recommendations for reporting caffeine and theophylline are still lacking. The aim of the study was to investigate IRLs for theobromine in blood and for caffeine and theophylline in blood and urine. Therefore, a set of six administrations were carried out including both single i.v. and single oral administrations of caffeine, theobromine and theophylline. Plasma and urine concentrations were determined using a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS). Applying the Toutain model approach an effective plasma concentration (EPC) of caffeine was estimated at 3.05 µg/mL, irrelevant concentrations in blood (IPC) and urine (IUC) approached 6 and 12 ng/mL, respectively. EPC of theobromine was calculated with 3.80 µg/mL, and irrelevant concentrations of theobromine were determined at 8 ng/mL in plasma and at 142 ng/mL in urine. Toutain modelling of the theophylline data produced an EPC, IPC, and IUC of 3.20 µg/mL, 6 ng/mL, and 75 ng/mL, respectively. The obtained irrelevant concentrations were used to postulate IRLs for theobromine in plasma and for caffeine and theophylline in plasma and urine. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  1. An acute intake of theobromine does not change postprandial lipid metabolism, whereas a high-fat meal lowers chylomicron particle number.

    Science.gov (United States)

    Smolders, Lotte; Mensink, Ronald P; Plat, Jogchum

    2017-04-01

    Postprandial responses predict cardiovascular disease risk. However, only a few studies have compared acute postprandial effects of a low-fat, high-carbohydrate (LF) meal with a high-fat, low-carbohydrate (HF) meal. Furthermore, theobromine has favorably affected fasting lipids, but postprandial effects are unknown. Because both fat and theobromine have been reported to increase fasting apolipoprotein A-I (apoA-I) concentrations, the main hypothesis of this randomized, double-blind crossover study was that acute consumption of an HF meal and a theobromine meal increased postprandial apoA-I concentrations, when compared with an LF meal. Theobromine was added to the LF meal. Nine healthy men completed the study. After meal intake, blood was sampled frequently for 4hours. Postprandial apoA-I concentrations were comparable after intake of the 3 meals. Apolipoprotein B48 curves, however, were significantly lower and those of triacylglycerol were significantly higher after HF as compared with LF consumption. Postprandial free fatty acid concentrations decreased less, and glucose and insulin concentrations increased less after HF meal consumption. Except for an increase in the incremental area under the curve for insulin, theobromine did not modify responses of the LF meal. These data show that acute HF and theobromine consumption does not change postprandial apoA-I concentrations. Furthermore, acute HF consumption had divergent effects on postprandial apolipoprotein B48 and triacylglycerol responses, suggesting the formation of less, but larger chylomicrons after HF intake. Finally, except for an increase in the incremental area under the curve for insulin, acute theobromine consumption did not modify the postprandial responses of the LF meal. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Simultaneous determination of theobromine, (+)-catechin, caffeine, and (-)-epicatechin in standard reference material baking chocolate 2384, cocoa, cocoa beans, and cocoa butter.

    Science.gov (United States)

    Risner, Charles H

    2008-01-01

    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 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, 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.

  3. Comparative pharmacokinetics of caffeine and its primary demethylated metabolites paraxanthine, theobromine and theophylline in man.

    Science.gov (United States)

    Lelo, A; Birkett, D J; Robson, R A; Miners, J O

    1986-01-01

    The pharmacokinetics of caffeine (CA), paraxanthine (PX), theobromine (TB) and theophylline (TP) were studied in six healthy male volunteers after oral administration of each compound on separate occasions. The total plasma clearances of CA and PX were similar in value (2.07 and 2.20 ml min-1 kg-1, respectively) as were those for TP and TB (0.93 and 1.20 ml min-1 kg-1, respectively). The unbound plasma clearances of CA and PX were also similar in magnitude (3.11 and 4.14 ml min-1 kg-1, respectively) as were those of TP and TB (1.61 and 1.39 ml min-1 kg-1, respectively). The half-lives of TP and TB (6.2 and 7.2 h, respectively) were significantly longer than those of CA and PX (4.1 and 3.1 h, respectively). The volume of distribution at steady state of TP (0.44 l kg-1) was lower than that of the other methylxanthines (0.63-0.72 l kg-1). The unbound volume of distribution of TP (0.77 l kg-1) was however the same as that of TB (0.79 l kg-1) whereas the unbound volume of distribution of PX (1.18 l kg-1) was similar to that of CA (1.06 l kg-1). PMID:3756065

  4. Study on the paper substrate room temperature phosphorescence of theobromine, caffeine and theophylline and analytical application

    Science.gov (United States)

    Chuan, Dong; Yan-Li, Wei; Shao-Min, Shuang

    2003-05-01

    Paper substrate room temperature phosphorescence (RTP) of theobromine (TB), caffeine (CF) and theophylline (TP) were investigated. The method is based on fast speed quantitative filter paper as substrate and KI-NaAc as heavy atom perturber. Various factors affecting their RTP were discussed in detail. Under the optimum experimental conditions, the linear dynamic range, limit of detection (LOD), and relative standard deviation (R.S.D.) were 14.41˜576.54 ng per spot, 1.14 ng per spot, 4.8% for TB, 5.44˜699.08 ng per spot, 0.78 ng per spot, 1.56% for CF, 7.21˜360.34 ng per spot, 1.80 ng per spot, 3.80% for TP, respectively. The first analytical application for the determination of these compounds was developed. The recovery of standard samples added to commercial products chocolate, tea, coffee and aminophylline is in the range 92.80-106.08%. The proposed method was successfully applied to real sample analysis without separation.

  5. The interaction of lysozyme with caffeine, theophylline and theobromine in solution.

    Science.gov (United States)

    Zhang, Hong-Mei; Tang, Bo-Ping; Wang, Yan-Qing

    2010-10-01

    The interactions of lysozyme with caffeine (Caf), theophylline (Tph) and theobromine (Tbr) were investigated using UV-Vis absorption, fluorescence, synchronous fluorescence, and three-dimensional fluorescence spectra techniques. The results revealed that Caf (Tph or Tbr) caused the fluorescence quenching of lysozyme by the formation of Caf (Tph or Tbr)-lysozyme complex. The binding constants (K(A)) and thermodynamic parameters (ΔG°, ΔH°, ΔS°) at two different temperatures, the binding locality, and the binding power were obtained. The results showed that the process of binding Caf (Tph or Tbr) to lysozyme was a spontaneous molecular interaction procedure and the hydrophobic and electrostatic interactions play a major role in stabilizing the complex; The distance r between donor (lysozyme) and acceptor (Caf, Tph or Tbr) was obtained according to fluorescence resonance energy transfer. The effect of Caf (Tph or Tbr) on the conformation of lysozyme was analyzed using synchronous fluorescence and three-dimensional fluorescence spectra techniques. The results showed that the binding of Caf (Tph or Tbr) to lysozyme induced some micro-environmental and conformational changes in lysozyme and disturbed the environment of the polypeptide of lysozyme.

  6. Quantification of theobromine and caffeine in saliva, plasma and urine via liquid chromatography-tandem mass spectrometry: a single analytical protocol applicable to cocoa intervention studies.

    Science.gov (United States)

    Ptolemy, Adam S; Tzioumis, Emma; Thomke, Arjun; Rifai, Sami; Kellogg, Mark

    2010-02-01

    Targeted analyses of clinically relevant metabolites in human biofluids often require extensive sample preparation (e.g., desalting, protein removal and/or preconcentration) prior to quantitation. In this report, a single ultra-centrifugation based sample pretreatment combined with a designed liquid chromatography-tandem mass spectrometry (LC-MS/MS) protocol provides selective quantification of 3,7-dimethylxanthine (theobromine) and 1,3,7-trimethylxanthine (caffeine) in human saliva, plasma and urine samples. The optimized chromatography permitted elution of both analytes within 1.3 min of the applied gradient. Positive-mode electrospray ionization and a triple quadruple MS/MS instrument operated in multiple reaction mode were used for detection. (13)C(3) isotopically labeled caffeine was included as an internal standard to improve accuracy and precision. Implementing a 20-fold dilution of the isolated low MW biofluid fraction prior to injection effectively minimized the deleterious contributions of all three matrices to quantitation. The assay was linear over a 160-fold concentration range from 2.5 to 400 micromol L(-1) for both theobromine (average R(2) 0.9968) and caffeine (average R(2) 0.9997) respectively. Analyte peak area variations for 2.5 micromol L(-1) caffeine and theobromine in saliva, plasma and urine ranged from 5 and 10% (intra-day, N=10) to 9 and 13% (inter-day, N=25) respectively. The intra- and inter-day precision of theobromine and caffeine elution times were 3 and theobromine ranged from 114 to 118% and 99 to 105% at concentration levels of 10 and 300 micromol L(-1). This validated protocol also permitted the relative saliva, plasma and urine distribution of both theobromine and caffeine to be quantified following a cocoa intervention. 2009 Elsevier B.V. All rights reserved.

  7. Theobromine does not affect postprandial lipid metabolism and duodenal gene expression, but has unfavorable effects on postprandial glucose and insulin responses in humans.

    Science.gov (United States)

    Smolders, Lotte; Mensink, Ronald P; Boekschoten, Mark V; de Ridder, Rogier J J; Plat, Jogchum

    2017-06-10

    Chocolate consumption is associated with a decreased risk for CVD. Theobromine, a compound in cocoa, may explain these effects as it favorably affected fasting serum lipids. However, long-term effects of theobromine on postprandial metabolism as well as underlying mechanisms have never been studied. The objective was to evaluate the effects of 4-week theobromine consumption (500 mg/day) on fasting and postprandial lipid, lipoprotein and glucose metabolism, and duodenal gene expression. In a randomized, double-blind crossover study, 44 healthy men and women, with low baseline HDL-C concentrations consumed 500 mg theobromine or placebo daily. After 4-weeks, fasting blood was sampled and subjects participated in a 4-h postprandial test. Blood was sampled frequently for analysis of lipid and glucose metabolism. In a subgroup of 10 men, 5 h after meal consumption duodenal biopsies were taken for microarray analysis. 4-weeks theobromine consumption lowered fasting LDL-C (-0.21 mmol/L; P = 0.006), and apoB100 (-0.04 g/L; P = 0.022), tended to increase HDL-C (0.03 mmol/L; P = 0.088) and increased hsCRP (1.2 mg/L; P = 0.017) concentrations. Fasting apoA-I, TAG, FFA, glucose and insulin concentrations were unchanged. In the postprandial phase, theobromine consumption increased glucose (P = 0.026), insulin (P = 0.011) and FFA (P = 0.003) concentrations, while lipids and (apo)lipoproteins were unchanged. In duodenal biopsies, microarray analysis showed no consistent changes in expression of genes, pathways or gene sets related to lipid, cholesterol or glucose metabolism. It is not likely that the potential beneficial effects of cocoa on CVD can be ascribed to theobromine. Although theobromine lowers serum LDL-C concentrations, it did not change fasting HDL-C, apoA-I, or postprandial lipid concentrations and duodenal gene expression, and unfavorably affected postprandial glucose and insulin responses. This trial was registered on clinicaltrials.gov under

  8. Spatial distribution of theobromine--a low MW drug--in tissues via matrix-free NALDI-MS imaging.

    Science.gov (United States)

    Tata, Alessandra; Montemurro, Chiara; Porcari, Andreia M; Silva, Kamila C; Lopes de Faria, José B; Eberlin, Marcos N

    2014-09-01

    The ability of nano-assisted laser desorption-ionization mass spectrometry imaging (NALDI-IMS) to provide selective chemical monitoring with appropriate spatial distribution of a low molecular drug in a biological tissue was investigated. NALDI-IMS is a matrix-free laser desorption ionization (LDI) protocol based on imprinting of tissue constituents on a nanostructured surface. Using the accumulation of theobromine in rat kidney as a model, NALDI-IMS was found to provide well-resolved images of the special distribution of this low molecular weight (MW) drug in tissue. Copyright © 2014 John Wiley & Sons, Ltd.

  9. Theobromine Upregulates Osteogenesis by Human Mesenchymal Stem Cells In Vitro and Accelerates Bone Development in Rats.

    Science.gov (United States)

    Clough, Bret H; Ylostalo, Joni; Browder, Elizabeth; McNeill, Eoin P; Bartosh, Thomas J; Rawls, H Ralph; Nakamoto, Tetsuo; Gregory, Carl A

    2017-03-01

    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

  10. In vitro transdermal delivery of caffeine, theobromine, theophylline and catechin from extract of Guarana, Paullinia Cupana.

    Science.gov (United States)

    Heard, Charles M; Johnson, Sarah; Moss, Gary; Thomas, Chris P

    2006-07-06

    Extracts of guarana (Paullinia cupana) feature as putatively stimulating ingredients in a number of foods, drinks and dietary/herbal supplements. The objective of this work was to investigate in vitro the transdermal delivery of the major pharmacologically active compounds contained in guarana extract. Saturated solutions of guarana were prepared in polyethylene glycol 400 (PEG400), propylene glycol (PG) and H(2)O at 32 degrees C. Guarana extract was also formulated in Duro-tak 2287 transdermal adhesive in a range of concentrations and the diffusional release was determined in addition to adhesive properties. Transdermal delivery across full thickness pig ear skin was investigated in vitro using Franz-type diffusion cells, with reverse-phase HPLC being used for the quantification of the permeation of theobromine (TB), theophylline (TP), (+)-catechin (C) and caffeine (CF). Based upon a combination of release and adhesive property data a patch containing 5.55 mg guarana extract cm(-2) was deemed optimal. The general trend for the delivery of the 4 analytes was: water >5.55 mg cm(-2) patch approximately PG>PEG400. For CF the greatest steady state flux was obtained from the water vehicle: 19 microg cm(-2)h(-1), with approximately 420 microg cm(-2) permeating after 24h. This was some 6x times more than from the drug-in-adhesive patch and 10x greater than PG, a well-known penetration enhancer, and 50x that of the 'regular' excipient PEG400. A water vehicle also provided the greatest delivery of TB (0.45 microg cm(-2) h(-1)), TP (0.022 microg cm(-2) h(-1)), and C (0.10 microg cm(-2) h(-1)). An inverse relationship was noted between lipophilicity and k(p) in each vehicle. The simultaneous transdermal delivery of the major actives of guarana was established, with permeation rates being highly concentration and vehicle dependent.

  11. Biochemical and ultrastructural changes in rabbit sclera after treatment with 7-methylxanthine, theobromine, acetazolamide, or L-ornithine

    Science.gov (United States)

    Trier, K.; Olsen, E. B.; Kobayashi, T.; Ribel-Madsen, S. M.

    1999-01-01

    AIMS—To examine a possible effect of 7-methylxanthine, theobromine, acetazolamide, or L-ornithine on the ultrastructure and biochemical composition of rabbit sclera.
METHODS—Groups of pigmented rabbits, six in each group, were dosed during 10 weeks with one of the substances under investigation, and one untreated group was the control. Samples of anterior and posterior sclera were taken for determination of hydroxyproline, hydroxylysine, proline, proteoglycans, uronic acids and dermatan sulphate, chondroitin sulphate, and hyaluronic acid. Sections were examined with electron microscopy, and the diameter of the individual collagen fibrils was measured.
RESULTS—Treatment with theobromine produced a significant increase in the contents of hydroxylysine, hydroxyproline, and proline in both anterior and posterior sclera, while 7-methylxanthine increased the contents of hydroxyproline and proline selectively in posterior sclera. Acetazolamide, on the other hand, significantly decreased the contents of hydroxyproline and proline in samples from anterior sclera. Uronic acids in both anterior and posterior sclera were significantly reduced by treatment with 7-methylxanthine, and L-ornithine significantly reduced uronic acids in posterior sclera. An inverse correlation between contents of hydroxyproline and uronic acids was found. The mean diameter of collagen fibrils was significantly higher in the posterior sclera from rabbits treated with 7-methylxanthine or theobromine, and significantly lower in rabbits treated with acetazolamide or L-ornithine compared with controls. In the anterior sclera, fibril diameter was significantly reduced in all treatment groups compared with controls. A positive, significant correlation between fibril diameter and content of hydroxyproline and proline was found in posterior sclera.
CONCLUSION—7-Methylxanthine, a metabolite of caffeine, increases collagen concentration and the diameter of collagen fibrils in the posterior

  12. Intermolecular Interactions in Crystalline Theobromine as Reflected in Electron Deformation Density and (13)C NMR Chemical Shift Tensors.

    Science.gov (United States)

    Bouzková, Kateřina; Babinský, Martin; Novosadová, Lucie; Marek, Radek

    2013-06-11

    An understanding of the role of intermolecular interactions in crystal formation is essential to control the generation of diverse crystalline forms which is an important concern for pharmaceutical industry. Very recently, we reported a new approach to interpret the relationships between intermolecular hydrogen bonding, redistribution of electron density in the system, and NMR chemical shifts (Babinský et al. J. Phys. Chem. A, 2013, 117, 497). Here, we employ this approach to characterize a full set of crystal interactions in a sample of anhydrous theobromine as reflected in (13)C NMR chemical shift tensors (CSTs). The important intermolecular contacts are identified by comparing the DFT-calculated NMR CSTs for an isolated theobromine molecule and for clusters composed of several molecules as selected from the available X-ray diffraction data. Furthermore, electron deformation density (EDD) and shielding deformation density (SDD) in the proximity of the nuclei involved in the proposed interactions are calculated and visualized. In addition to the recently reported observations for hydrogen bonding, we focus here particularly on the stacking interactions. Although the principal relations between the EDD and CST for hydrogen bonding (HB) and stacking interactions are similar, the real-space consequences are rather different. Whereas the C-H···X hydrogen bonding influences predominantly and significantly the in-plane principal component of the (13)C CST perpendicular to the HB path and the C═O···H hydrogen bonding modulates both in-plane components of the carbonyl (13)C CST, the stacking modulates the out-of-plane electron density resulting in weak deshielding (2-8 ppm) of both in-plane principal components of the CST and weak shielding (∼ 5 ppm) of the out-of-plane component. The hydrogen-bonding and stacking interactions may add to or subtract from one another to produce total values observed experimentally. On the example of theobromine, we demonstrate

  13. A theobromine derived silver N-heterocyclic carbene: synthesis, characterization, and antimicrobial efficacy studies on cystic fibrosis relevant pathogens.

    Science.gov (United States)

    Panzner, Matthew J; Hindi, Khadijah M; Wright, Brian D; Taylor, Jane B; Han, Daniel S; Youngs, Wiley J; Cannon, Carolyn L

    2009-09-21

    The increasing incidence of multidrug-resistant (MDR) pulmonary infections in the cystic fibrosis (CF) population has prompted the investigation of innovative silver based therapeutics. The functionalization of the naturally occurring xanthine theobromine at the N(1) nitrogen atom with an ethanol substituent followed by the methylation of the N(9) nitrogen atom gives the N-heterocyclic carbene precursor 1-(2-hydroxyethyl)-3,7,9-trimethylxanthinium iodide. The reaction of this xanthinium salt with silver acetate produces the highly hydrophilic silver carbene complex SCC8. The in vitro antimicrobial efficacy of this newly synthesized complex was evaluated with excellent results on a variety of virulent and MDR pathogens isolated from CF patients. A comparative in vivo study between the known caffeine derived silver carbene SCC1 and SCC8 demonstrated the ability of both complexes to improve the survival rates of mice in a pneumonia model utilizing the clinically isolated infectious strain of Pseudomonas aeruginosa PA M57-15.

  14. Simultaneous multiplexed quantification of caffeine and its major metabolites theobromine and paraxanthine using surface-enhanced Raman scattering.

    Science.gov (United States)

    Alharbi, Omar; Xu, Yun; Goodacre, Royston

    2015-11-01

    Accurate quantitative measurement of drugs and their metabolites is important as this can be used to establish long-term abuse of illicit materials as well as establish accurate drug dosing for legal therapeutics. However, the levels of drugs and xenometabolites found in human body fluids necessitate methods that are highly sensitive as well as reproducible with the potential for portability. Raman spectroscopy does offer excellent reproducibility, portability and chemical specificity, but unfortunately, the Raman effect is generally too weak unless it is enhanced. We therefore developed surface-enhanced Raman scattering (SERS) and combined it with the powerful machine learning technique of artificial neural networks to enable rapid quantification of caffeine and its two major metabolites theobromine and paraxanthine. We established a three-way mixture analysis from 10(-5) to 10(-7) mol/dm(3), and excellent predictions were generated for all three analytes in tertiary mixtures. The range we selected reflects the levels found in human body fluids, and the typical errors for our portable SERS analysis were 1.7 × 10(-6) mol/dm(3) for caffeine, 8.8 × 10(-7) mol/dm(3) for theobromine and 9.6 × 10(-7) mol/dm(3) for paraxanthine. We believe this demonstrates the exciting prospect of using SERS for the quantitative analysis of multiple analytes simultaneously without recourse to lengthy and time-consuming chromatography, a method that often has to be combined with mass spectrometry.

  15. Theobromine-Induced Changes in A1 Purinergic Receptor Gene Expression and Distribution in a Rat Brain Alzheimer's Disease Model.

    Science.gov (United States)

    Mendiola-Precoma, Jesus; Padilla, Karla; Rodríguez-Cruz, Alfredo; Berumen, Laura C; Miledi, Ricardo; García-Alcocer, Guadalupe

    2017-01-01

    Dementia caused by Alzheimer's disease (AD) is mainly characterized by accumulation in the brain of extra- and intraneuronal amyloid-β (Aβ) and tau proteins, respectively, which selectively affect specific regions, particularly the neocortex and the hippocampus. Sporadic AD is mainly caused by an increase in apolipoprotein E, a component of chylomicrons, which are cholesterol transporters in the brain. Recent studies have shown that high lipid levels, especially cholesterol, are linked to AD. Adenosine is an atypical neurotransmitter that regulates a wide range of physiological functions by activating four P1 receptors (A1, A2A, A2B, and A3) and P2 purinergic receptors that are G protein-coupled. A1 receptors are involved in the inhibition of neurotransmitter release, which could be related to AD. The aim of the present work was to study the effects of a lard-enriched diet (LED) on cognitive and memory processes in adult rats (6 months of age) as well as the effect of theobromine on these processes. The results indicated that the fat-enriched diet resulted in a long-term deterioration in cognitive and memory functions. Increased levels of Aβ protein and IL-1β were also observed in the rats fed with a high-cholesterol diet, which were used to validate the AD animal model. In addition, the results of qPCR and immunohistochemistry indicated a decrease in gene expression and distribution of A1 purinegic receptor, respectively, in the hippocampus of LED-fed rats. Interestingly, theobromine, at both concentrations tested, restored A1 receptor levels and improved cognitive functions and Aβ levels for a dose of 30 mg/L drinking water.

  16. Determination of caffeine, theobromine and theophylline in Mate beer and Mate soft drinks by high-performance thin-layer chromatography.

    Science.gov (United States)

    Oellig, Claudia; Schunck, Jacob; Schwack, Wolfgang

    2018-01-19

    Mate beer and Mate soft drinks are beverages produced from the dried leaves of Ilex paraguariensis (Yerba Mate). In Yerba Mate, the xanthine derivatives caffeine, theobromine and theophylline, also known as methylxanthines, are important active components. The presented method for the determination of caffeine, theobromine and theophylline in Mate beer and Mate soft drinks by high-performance thin-layer chromatography with ultraviolet detection (HPTLC-UV) offers a fully automated and sensitive determination of the three methylxanthines. Filtration of the samples was followed by degassing, dilution with acetonitrile in the case of Mate beers for protein precipitation, and centrifugation before the extracts were analyzed by HPTLC-UV on LiChrospher silica gel plates with fluorescence indicator and acetone/toluene/chloroform (4:3:3, v/v/v) as the mobile phase. For quantitation, the absorbance was scanned at 274nm. Limits of detection and quantitation were 1 and 4ng/zone, respectively, for caffeine, theobromine and theophylline. With recoveries close to 100% and low standard deviations reliable results were guaranteed. Experimental Mate beers as well as Mate beers and Mate soft drinks from the market were analyzed for their concentrations of methylxanthines. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Collaborative work to evaluate toxicity on male reproductive organs by repeated dose studies in rats 22). Effects of 2- and 4-week administration of theobromine on the testis.

    Science.gov (United States)

    Funabashi, H; Fujioka, M; Kohchi, M; Tateishi, Y; Matsuoka, N

    2000-10-01

    The effects of theobromine, a xanthine derivative, on the testis were compared between rats dosed for 2 and 4 weeks to determine whether a 2-week dosing period is long enough to detect toxicity. Theobromine was administered orally to male Sprague-Dawley rats at dose levels of 250 and 500 mg/kg for 2 weeks starting at the age of 6 or 8 weeks, and for 4 weeks from the age of 6 weeks. Histopathological examination of reproductive organs revealed toxic findings in the testis at 500 mg/kg after 2 weeks of dosing at both ages, and at 250 and 500 mg/kg after 4 weeks of dosing. The primary findings were degeneration/necrosis and desquamation of spermatids and spermatocytes, vacuolization of seminiferous tubules, and multinucleated giant cell formation. These findings were present mainly in stages I-VI and XII-XIV. From these results, it is concluded that the toxic effects of theobromine on the testis can be detected by repeated dosing for 2 weeks as well as for 4 weeks.

  18. Direct injection liquid chromatography/electrospray ionization mass spectrometric horse urine analysis for the quantification and confirmation of threshold substances for doping control. II. Determination of theobromine.

    Science.gov (United States)

    Vonaparti, A; Lyris, E; Panderi, I; Koupparis, M; Georgakopoulos, C

    2009-04-01

    In equine sport, theobromine is prohibited with a threshold level of 2 microg mL(-1) in urine, hence doping control laboratories have to establish quantitative and qualitative methods for its determination. Two simple liquid chromatography/mass spectrometry (LC/MS) methods for the identification and quantification of theobromine were developed and validated using the same sample preparation procedure but different mass spectrometric systems: ion trap mass spectrometry (ITMS) and time-of-flight mass spectrometry (TOFMS). Particle-free diluted urine samples were directly injected into the LC/MS systems, avoiding the time-consuming extraction step. 3-Propylxanthine was used as the internal standard. The tested linear range was 0.75-15 microg mL(-1). Matrix effects were evaluated analyzing calibration curves in water and different fortified horse urine samples. A great variation in the signal of theobromine and the internal standard was observed in different matrices. To overcome matrix effects, a standard additions calibration method was applied. The relative standard deviations of intra- and inter-day analysis were lower than 8.6 and 7.2%, respectively, for the LC/ITMS method and lower than 5.7 and 5.8%, respectively, for the LC/TOFMS method. The bias was less than 8.7% for both methods. The methods were applied to two case samples, demonstrating simplicity, accuracy and selectivity. Copyright (c) 2009 John Wiley & Sons, Ltd.

  19. A theobromine derived silver N-heterocyclic carbene: synthesis, characterization, and antimicrobial efficacy studies on cystic fibrosis relevant pathogens†

    Science.gov (United States)

    Panzner, Matthew J.; Hindi, Khadijah M.; Wright, Brian D.; Taylor, Jane B.; Han, Daniel S.

    2009-01-01

    The increasing incidence of multidrug-resistant (MDR) pulmonary infections in the cystic fibrosis (CF) population has prompted the investigation of innovative silver based therapeutics. The functionalization of the naturally occurring xanthine theobromine at the N1 nitrogen atom with an ethanol substituent followed by the methylation of the N9 nitrogen atom gives the N-heterocyclic carbene precursor 1-(2-hydroxyethyl)-3,7,9-trimethylxanthinium iodide. The reaction of this xanthinium salt with silver acetate produces the highly hydrophilic silver carbene complex SCC8. The in vitro antimicrobial efficacy of this newly synthesized complex was evaluated with excellent results on a variety of virulent and MDR pathogens isolated from CF patients. A comparative in vivo study between the known caffeine derived silver carbene SCC1 and SCC8 demonstrated the ability of both complexes to improve the survival rates of mice in a pneumonia model utilizing the clinically isolated infectious strain of Pseudomonas aeruginosa PA M57-15. PMID:20449175

  20. Simultaneous determination of artificial sweeteners, preservatives, caffeine, theobromine and theophylline in food and pharmaceutical preparations by ion chromatography.

    Science.gov (United States)

    Chen, Q C; Wang, J

    2001-12-07

    A novel ion chromatographic method was proposed for the simultaneous determination of artificial sweeteners (sodium saccharin, aspartame, acesulfame-K), preservatives (benzoic acid, sorbic acid), caffeine, theobromine and theophylline. The separation was performed on an anion-exchange analytical column operated at 40 degrees C within 45 min by an isocratic elution with 5 mM aqueous NaH2PO4 (pH 8.20) solution containing 4% (v/v) acetonitrile as eluent, and the determination by wavelength-switching ultraviolet absorbance detection. The detection limits (signal-to-noise ratio 3:1) for all analytes were below the sub-microg/ml level. Under the experimental conditions, several organic acids, including citric acid, malic acid, tartaric acid and ascorbic acid, did not interfere with the determination. The method has been successfully applied to the analysis of various food and pharmaceutical preparations, and the average recoveries for real samples ranged from 85 to 104%. The levels of all analytes determined by this method were in good agreement with those obtained by the high-performance liquid chromatographic procedure. The results also indicated that ion chromatography would be possibly a beneficial alternative to conventional high-performance liquid chromatography for the separation and determination of these compounds.

  1. Topology of the interactions pattern in pharmaceutically relevant polymorphs of methylxanthines (caffeine, theobromine, and theophiline): combined experimental (¹H-¹⁴N nuclear quadrupole double resonance) and computational (DFT and Hirshfeld-based) study.

    Science.gov (United States)

    Latosińska, Jolanta Natalia; Latosińska, Magdalena; Olejniczak, Grzegorz A; Seliger, Janez; Žagar, Veselko

    2014-09-22

    Three anhydrous methylxanthines: caffeine (1,3,7-trimethylxanthine; 1,3,7-trimethyl-1H-purine-2,6-(3H,7H)-dione) and its two metabolites theophylline (1,3-dimethylxanthine; 1,3-dimethyl-7H-purine-2,6-dione) and theobromine (3,7-dimethyl-xanthine; 3,7-dimethyl-7H-purine-2,6-dione), which reveal multifaceted therapeutic potential, have been studied experimentally in solid state by (1)H-(14)N NMR-NQR (nuclear magnetic resonance-nuclear quadrupole resonance) double resonance (NQDR). For each compound the complete NQR spectrum consisting of 12 lines was recorded. The multiplicity of NQR lines indicates the presence of a stable β form of anhydrous caffeine at 233 K and stable form II of anhydrous theobromine at 213 K. The assignment of signals detected in NQR experiment to particular nitrogen atoms was made on the basis of quantum chemistry calculations performed for monomer, cluster, and solid at the DFT/GGA/BLYP/DPD level. The shifts due to crystal packing interactions were evaluated, and the multiplets detected by NQR were assigned to N(9) in theobromine and N(1) and N(9) in caffeine. The ordering theobromine > theophylline > caffeine site and theophylline theobromine theobromine) to π···π stacking (caffeine). Substantial differences in the intermolecular interactions in stable forms of methylxanthines differing in methylation (site or number) were analyzed within the Hirshfeld surface-based approach. The analysis of local environment of the nitrogen nucleus permitted drawing some conclusions on the nature of the interactions required for effective processes of recognition and binding of a given methylxanthine to A1-A(2A) receptor (target for caffeine in the brain). Although the interactions responsible for linking neighboring methylxanthines molecules in crystals and methylxanthines with targets in the human organism can differ significantly, the knowledge of the topology of interactions provides reliable preliminary information about the nature of this binding.

  2. Application of RNAi to confirm theobromine as the major intermediate for caffeine biosynthesis in coffee plants with potential for construction of decaffeinated varieties.

    Science.gov (United States)

    Ogita, Shinjiro; Uefuji, Hirotaka; Morimoto, Masayuki; Sano, Hiroshi

    2004-04-01

    The caffeine biosynthetic pathway in coffee plants has been proposed to involve three distinct N -methyltransferases, xanthosine methyltransferase (XMT), 7- N -methylxanthine methyltransferase (MXMT; theobromine synthase), and 3,7-dimethylxanthine methyltransferase (DXMT; caffeine synthase). We previously isolated all corresponding cDNAs designated as CaXMT1 , CaMXMT1 , CaMXMT2 and CaDXMT1 , respectively, and showed that caffeine was indeed synthesized in vitro by the combination of their gene products. In order to regulate caffeine biosynthesis in planta , we suppressed expression of CaMXMT1 by the double stranded RNA interference (RNAi) method. For this purpose, we first established a protocol for efficient somatic embryogenesis of Coffea arabica and C. canephora , and then Agrobacterium -mediated transformation techniques. The RNAi transgenic lines of embryogenic tissues derived from C. arabica and transgenic plantlets of C. canephora demonstrated a clear reduction in transcripts for CaMXMT1 in comparison with the control plants. Transcripts for CaXMT1 and CaDXMT1 were also reduced in the most cases. Both embryonic tissues and plantlets exhibited a concomitant reduction of theobromine and caffeine contents to a range between 30% and 50% of that of the control. These results suggest that the CaMXMT1 -RNAi sequence affected expression of not only CaMXMT1 itself, but also CaXMT1 and CaDXMT1 , and that, since the reduction in theobromine content was proportional to that for caffeine, it is involved in the major synthetic pathway in coffee plants. The results also indicate that the method can be practically applied to produce decaffeinated coffee plants.

  3. Rapid methods to determine procyanidins, anthocyanins, theobromine and caffeine in rat tissues by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Serra, Aida; Macià, Alba; Romero, Maria-Paz; Piñol, Carme; Motilva, Maria-José

    2011-06-01

    Rapid, selective and sensitive methods were developed and validated to determine procyanidins, anthocyanins and alkaloids in different biological tissues, such as liver, brain, the aorta vein and adipose tissue. For this purpose, standards of procyanidins (catechin, epicatechin, and dimer B(2)), anthocyanins (cyanidin-3-glucoside and malvidin-3-glucoside) and alkaloids (theobromine, caffeine and theophylline) were used. The methods included the extraction of homogenized tissues by off-line liquid-solid extraction, and then solid-phase extraction to analyze alkaloids, or microelution solid-phase extraction plate for the analysis of procyanidins and anthocyanins. The eluted extracts were then analyzed by ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry, using a triple quadrupole as the analyzer. The optimum extraction solution was water/methanol/phosphoric acid 4% (94/4.5/1.5, v/v/v). The extraction recoveries were higher than 81% for all the studied compounds in all the tissues, except the anthocyanins, which were between 50 and 65% in the liver and brain. In order to show the applicability of the developed methods, different rat tissues were analyzed to determine the procyanidins, anthocyanins and alkaloids and their generated metabolites. The rats had previously consumed 1g of a grape pomace extract (to analyze procyanidins and anthocyanins) or a cocoa extract (to analyze alkaloids) per kilogram of body weight. Different tissues were extracted 4h after administration of the respective extracts. The analysis of the metabolites revealed a hepatic metabolism of procyanidins. The liver was the tissue which produced a greater accumulation of these metabolites. Copyright © 2011 Elsevier B.V. All rights reserved.

  4. Population pharmacokinetics of caffeine and its metabolites theobromine, paraxanthine and theophylline after inhalation in combination with diacetylmorphine.

    Science.gov (United States)

    Zandvliet, Anthe S; Huitema, Alwin D R; de Jonge, Milly E; den Hoed, Rob; Sparidans, Rolf W; Hendriks, Vincent M; van den Brink, Wim; van Ree, Jan M; Beijnen, Jos H

    2005-01-01

    The stimulant effect of caffeine, as an additive in diacetylmorphine preparations for study purposes, may interfere with the pharmacodynamic effects of diacetylmorphine. In order to obtain insight into the pharmacology of caffeine after inhalation in heroin users, the pharmacokinetics of caffeine and its dimethylxanthine metabolites were studied. The objectives were to establish the population pharmacokinetics under these exceptional circumstances and to compare the results to published data regarding intravenous and oral administration in healthy volunteers. Diacetylmorphine preparations containing 100 mg of caffeine were used by 10 persons by inhalation. Plasma concentrations of caffeine, theobromine, paraxanthine and theophylline were measured by high performance liquid chromatography. Non-linear mixed effects modelling was used to estimate population pharmacokinetic parameters. The model was evaluated by the jack-knife procedure. Caffeine was rapidly and effectively absorbed after inhalation. Population pharmacokinetics of caffeine and its dimethylxanthine metabolites could adequately and simultaneously be described by a linear multi-compartment model. The volume of distribution for the central compartment was estimated to be 45.7 l and the apparent elimination rate constant of caffeine at 8 hr after inhalation was 0.150 hr(-1) for a typical individual. The bioavailability was approximately 60%. The presented model adequately describes the population pharmacokinetics of caffeine and its dimethylxanthine metabolites after inhalation of the caffeine sublimate of a 100 mg tablet. Validation proved the stability of the model. Pharmacokinetics of caffeine after inhalation and intravenous administration are to a large extent similar. The bioavailability of inhaled caffeine is approximately 60% in experienced smokers.

  5. Analysis of Theobromine and Related Compounds by Reversed Phase High-Performance Liquid Chromatography with Ultraviolet Detection: An Update (1992–2011

    Directory of Open Access Journals (Sweden)

    Sandra Aparecida de Assis

    2011-01-01

    Full Text Available Theobromine and its related compounds, such as caffeine and theophylline, are secondary metabolites that belong to the alkaloids and have economic and cultural importance. These alkaloids have demonstrated stimulatory effects on the central nervous, gastrointestinal, cardiovascular, renal and respiratory systems, resulting in 'energy arousal', increased motivation to work, increased alertness and increased cognitive function. Several analytical methods have been used to analyse these compounds, but reversed phase high-performance liquid chromatography (RP-HPLC is the most commonly applied because of its efficiency, sensitivity, specificity and speed. This review describes the analyses of theobromine-related compounds by RP-HPLC with ultraviolet detection (UV in four sources: food, beverages, biological fluids and plants. Many RP-HPLC methods have been developed and optimized for the detection and quantification of these natural compounds. Elution under isocratic conditions is the most frequent method, with a water, methanol and acetonitrile mixture modified with acetic, phosphoric or formic acid as the mobile phase. For xanthine analysis, the use of reversed phase high-performance liquid chromatography with an ultraviolet/diode array detector (UV/DAD is particularly suitable as derivation is not required; it allows the analysis of absorbance at all wavelengths, it is simple and rapid.

  6. Determination of theobromine, theophylline, and caffeine in by-products of cupuacu and cacao seeds by high-performance liquid chromatography.

    Science.gov (United States)

    Lo Coco, F; Lanuzza, F; Micali, G; Cappellano, G

    2007-01-01

    Theobromine, theophylline, and caffeine are determined simultaneously by a rapid and selective reversed-phase high-performance liquid chromatography (HPLC) method with UV detection in by-products of cupuacu and cacao seeds. The determination is carried out in the raw and roasted ground cupuacu seeds and in the corresponding powders obtained after pressure treatment. The by-products of both cupuacu seeds and cacao seeds are obtained under the same technological conditions. The HPLC method uses isocratic elution with a mobile phase of methanol-water-acetic acid (80:19:1) (v/v) at a flow rate of 1 mL/min and UV absorbance detection at 275 nm. Total elution time for these analytes is less than 10 min, and the detection limit for all analytes is 0.1 mg/g. The amounts of theobromine and caffeine found in all the cupuacu samples are one or more orders of magnitude lower than those from cacao. Theophylline is found in all cacao samples except for the roasted ground paste, and it is only found in the roasted ground paste in the cupuacu samples.

  7. Theobromine, the primary methylxanthine found in Theobroma cacao, prevents malignant glioblastoma proliferation by negatively regulating phosphodiesterase-4, extracellular signal-regulated kinase, Akt/mammalian target of rapamycin kinase, and nuclear factor-kappa B.

    Science.gov (United States)

    Sugimoto, Naotoshi; Miwa, Shinji; Hitomi, Yoshiaki; Nakamura, Hiroyuki; Tsuchiya, Hiroyuki; Yachie, Akihiro

    2014-01-01

    Theobromine, a caffeine derivative, is the primary methylxanthine produced by Theobroma cacao. We previously showed that methylxanthines, including caffeine and theophylline, have antitumor and antiinflammatory effects, which are in part mediated by their inhibition of phosphodiesterase (PDE). A member of the PDE family, PDE4, is widely expressed in and promotes the growth of glioblastoma, the most common type of brain tumor. The purpose of this study was to determine whether theobromine could exert growth inhibitory effects on U87-MG, a cell line derived from human malignant glioma. We show that theobromine treatment elevates intracellular cAMP levels and increases the activity of p38 mitogen-activated protein kinase and c-Jun N-terminal kinase, whereas it attenuates p44/42 extracellular signal-regulated kinase activity and the Akt/mammalian target of rapamycin kinase and nuclear factor-kappa B signal pathways. It also inhibits cell proliferation. These results suggest that foods and beverages containing cocoa bean extracts, including theobromine, might be extremely effective in preventing human glioblastoma.

  8. Evaluation of the perinatal, postnatal and teratogenic effects of cocoa powder and theobromine in Sprague-Dawley/CD rats.

    Science.gov (United States)

    Tarka, S M; Applebaum, R S; Borzelleca, J F

    1986-05-01

    Studies were conducted to evaluate the effects of cocoa powder (CP) and theobromine (TBR) on perinatal and postnatal parameters and to assess their potential teratogenicity in the rat. In the peri/postnatal study, CP was given at 0, 2.5, 5.0 or 7.5% in the diet throughout gestation and lactation (postnatal day 21). In the teratology study, rats were given diets containing 0, 2.5 or 5.0% CP or 0.0675 or 0.135% TBR on days 6-19 of gestation. The CP-treated dams in the peri/postnatal study consumed significantly more food than did the controls during gestation. Weight gain was increased only in the 5.0 and 7.5% CP groups during lactation. Litter size was reduced slightly at 7.5% CP and pup survival was slightly decreased at 5.0 and 7.5% CP but none of these reductions was statistically significant. However, small but statistically significant decreases in pup body weights were noted in all treatment groups throughout lactation. In the teratology studies, rats given 2.5 or 5.0% CP or 0.0675 or 0.135% TBR consumed significantly more food than did the controls and the CP-treated dams gained significantly more weight. The percentage of pregnant dams and the mean number of corpora lutea were not affected by either CP or TBR. Foetuses exposed to 0.135 TBR had a significantly higher incidence of incompletely ossified or absent sternebrae and pubic bones, indicating a delay in osteogenesis. On the basis of the survival of treated offspring in the peri/postnatal study, these effects were not considered to be deleterious to either growth or survival. The effects are similar to those that have been reported elsewhere and been considered to indicate potential maternal or foetal toxicity that is unrelated to a specific compound/treatment. We conclude that any variations observed in these studies may be attributed to this non-specific maternal toxicity and are not related to the ingestion of either CP or TBR. The major methylxanthine found in the serum after CP or TBR ingestion was

  9. Synthesis and anticancer activity of silver(I)-N-heterocyclic carbene complexes derived from the natural xanthine products caffeine, theophylline and theobromine.

    Science.gov (United States)

    Mohamed, Heba A; Lake, Benjamin R M; Laing, Thomas; Phillips, Roger M; Willans, Charlotte E

    2015-04-28

    A new library of silver(I)-N-heterocyclic carbene complexes prepared from the natural products caffeine, theophylline and theobromine is reported. The complexes have been fully characterised using a combination of NMR spectroscopy, mass spectrometry, elemental analysis and X-ray diffraction analysis. Furthermore, the hydrophobicity of the complexes has been measured. The silver(I)-N-heterocyclic carbenes have been evaluated for their antiproliferative properties against a range of cancer cell lines of different histological types, and compared to cisplatin. The data shows different profiles of response when compared to cisplatin in the same panel of cells, indicating a different mechanism of action. Furthermore, it appears that the steric effect of the ligand and the hydrophobicity of the complex both play a role in the chemosensitivity of these compounds, with greater steric bulk and greater hydrophilicity delivering higher cytotoxicity.

  10. Molecular structures and vibrational frequencies of xanthine and its methyl derivatives (caffeine and theobromine) by ab initio Hartree-Fock and density functional theory calculations

    Science.gov (United States)

    Ucun, Fatih; Sağlam, Adnan; Güçlü, Vesile

    2007-06-01

    The molecular structures, vibrational frequencies and corresponding vibrational assignments of xanthine and its methyl derivatives (caffeine and theobromine) have been calculated using ab initio Hartree-Fock (HF) and density functional theory (B3LYP) methods with 6-31G(d, p) basis set level. The calculations were utilized to the CS symmetries of the molecules. The obtained vibrational frequencies and optimised geometric parameters (bond lengths and bond angles) were seen to be well agreement with the experimental data. The used scale factors which have been obtained the ratio of the frequency values of the strongest peaks in the calculated and experimental spectra seem to cause the gained vibrations well corresponding to the experimental ones. Theoretical infrared intensities and Raman activities are also reported.

  11. Magnetic Solid-phase Extraction with Fe3O4/Molecularly Imprinted Polymers Modified by Deep Eutectic Solvents and Ionic Liquids for the Rapid Purification of Alkaloid Isomers (Theobromine and Theophylline from Green Tea

    Directory of Open Access Journals (Sweden)

    Guizhen Li

    2017-06-01

    Full Text Available Different kinds of deep eutectic solvents (DES based on choline chloride (ChCl and ionic liquids (ILs based on 1-methylimidazole were used to modify Fe3O4/molecularly imprinted polymers (Fe3O4/MIPs, and the resulting materials were applied for the rapid purification of alkaloid isomers (theobromine and theophylline from green tea with magnetic solid-phase extraction (M-SPE. The M-SPE procedure was optimized using the response surface methodology (RSM to analyze the maximum conditions. The materials were characterized by Fourier transform infrared spectroscopy (FI-IR and field emission scanning electron microscopy (FE-SEM. Compared to the ILs-Fe3O4/MIPs, the DESs-Fe3O4/MIPs were developed for the stronger recognition and higher recoveries of the isomers (theophylline and theobromine from green tea, particularly DES-7-Fe3O4/MIPs. With RSM, the optimal recovery condition for theobromine and theophylline in the M-SPE were observed with ratio of methanol (80% as the washing solution, methanol/acetic acid (HAc (8:2 as the eluent at pH 3, and an eluent volume of 4 mL. The practical recoveries of theobromine and theophylline in green tea were 92.27% and 87.51%, respectively, with a corresponding actual extraction amount of 4.87 mg•g−1 and 5.07 mg•g−1. Overall, the proposed approach with the high affinity of Fe3O4/MIPs might offer a novel method for the purification of complex isomer samples.

  12. Magnetic Solid-phase Extraction with Fe₃O₄/Molecularly Imprinted Polymers Modified by Deep Eutectic Solvents and Ionic Liquids for the Rapid Purification of Alkaloid Isomers (Theobromine and Theophylline) from Green Tea.

    Science.gov (United States)

    Li, Guizhen; Wang, Xiaoqin; Row, Kyung Ho

    2017-06-25

    Different kinds of deep eutectic solvents (DES) based on choline chloride (ChCl) and ionic liquids (ILs) based on 1-methylimidazole were used to modify Fe3O4/molecularly imprinted polymers (Fe3O4/MIPs), and the resulting materials were applied for the rapid purification of alkaloid isomers (theobromine and theophylline) from green tea with magnetic solid-phase extraction (M-SPE). The M-SPE procedure was optimized using the response surface methodology (RSM) to analyze the maximum conditions. The materials were characterized by Fourier transform infrared spectroscopy (FI-IR) and field emission scanning electron microscopy (FE-SEM). Compared to the ILs-Fe3O4/MIPs, the DESs-Fe3O4/MIPs were developed for the stronger recognition and higher recoveries of the isomers (theophylline and theobromine) from green tea, particularly DES-7-Fe3O4/MIPs. With RSM, the optimal recovery condition for theobromine and theophylline in the M-SPE were observed with ratio of methanol (80%) as the washing solution, methanol/acetic acid (HAc) (8:2) as the eluent at pH 3, and an eluent volume of 4 mL. The practical recoveries of theobromine and theophylline in green tea were 92.27% and 87.51%, respectively, with a corresponding actual extraction amount of 4.87 mg•g-1 and 5.07 mg•g-1. Overall, the proposed approach with the high affinity of Fe3O4/MIPs might offer a novel method for the purification of complex isomer samples.

  13. Simultaneous determination of caffeine, theophylline and theobromine in human plasma by on-line solid-phase extraction coupled to reversed-phase chromatography.

    Science.gov (United States)

    Emara, Samy

    2004-10-01

    A reversed-phase liquid chromatographic column switching system was described for the determination of caffeine (CF), theophylline (TH) and theobromine (TB) in human plasma with a direct injection procedure. A short protein-coated mu Bondapak CN silica pre-column (20 x 3 mm, i.d.) was used for enrichment of the drugs and clean up from weakly retained plasma components using phosphate buffer saline pH 7.4. After washing step, the retained drugs were flushed into a reversed-phase column (5 microm TSK gel ODS-80 TM, 150 x 4.6 mm i.d.) with a mobile phase of methanol-0.01 M phosphate buffer, pH 3.5 (30:70, v/v) for the final separation. The eluent was monitored with a UV detector at 275 nm. The resulting chromatograms showed no interference from endogenous plasma components. A linear relationship between the concentration of drug and peak height was confirmed in the range of 0.5-20 microg/mL for all drugs. High extraction recoveries from plasma ranging from 96.12 to 100.32% were achieved. Validation of the method was examined performing intra- and inter-day accuracy and precision and was found to be satisfactory. The coefficients of variation of the three drugs were less than 3% for intra-day and less than 4% for inter-day run assays.

  14. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of health claims related to caffeine and theobromine in cocoa (Theobroma cacao L.) and enhancement of mood (ID 4276) pursuant to Article 13(1) of Regulation (EC) No 1924/2006

    DEFF Research Database (Denmark)

    Tetens, Inge

    or directly from stakeholders. The food that is the subject of the health claim is cocoa (Theobroma cacao L.). The Panel considers that whereas the food, cocoa (Theobroma cacao L.), is not sufficiently characterised in relation to the claimed effect evaluated in this opinion, the food constituents, caffeine...... that a cause and effect relationship has not been established between the consumption of caffeine and theobromine in cocoa (Theobroma cacao L.) and enhancement of mood....... claims in relation to caffeine and theobromine in cocoa (Theobroma cacao L.) and enhancement of mood. The scientific substantiation is based on the information provided by the Member States in the consolidated list of Article 13 health claims and references that EFSA has received from Member States...

  15. Validity and reproducibility of a self-administered semi-quantitative food-frequency questionnaire for estimating usual daily fat, fibre, alcohol, caffeine and theobromine intakes among Belgian post-menopausal women.

    Science.gov (United States)

    Bolca, Selin; Huybrechts, Inge; Verschraegen, Mia; De Henauw, Stefaan; Van de Wiele, Tom

    2009-01-01

    A novel food-frequency questionnaire (FFQ) was developed and validated to assess the usual daily fat, saturated, mono-unsaturated and poly-unsaturated fatty acid, fibre, alcohol, caffeine, and theobromine intakes among Belgian post-menopausal women participating in dietary intervention trials with phyto-oestrogens. The relative validity of the FFQ was estimated by comparison with 7 day (d) estimated diet records (EDR, n 64) and its reproducibility was evaluated by repeated administrations 6 weeks apart (n 79). Although the questionnaire underestimated significantly all intakes compared to the 7 d EDR, it had a good ranking ability (r 0.47-0.94; weighted kappa 0.25-0.66) and it could reliably distinguish extreme intakes for all the estimated nutrients, except for saturated fatty acids. Furthermore, the correlation between repeated administrations was high (r 0.71-0.87) with a maximal misclassification of 7% (weighted kappa 0.33-0.80). In conclusion, these results compare favourably with those reported by others and indicate that the FFQ is a satisfactorily reliable and valid instrument for ranking individuals within this study population.

  16. Validity and Reproducibility of a Self-Administered Semi-Quantitative Food-Frequency Questionnaire for Estimating Usual Daily Fat, Fibre, Alcohol, Caffeine and Theobromine Intakes among Belgian Post-Menopausal Women

    Directory of Open Access Journals (Sweden)

    Selin Bolca

    2009-01-01

    Full Text Available A novel food-frequency questionnaire (FFQ was developed and validated to assess the usual daily fat, saturated, mono-unsaturated and poly-unsaturated fatty acid, fibre, alcohol, caffeine, and theobromine intakes among Belgian post-menopausal women participating in dietary intervention trials with phyto-oestrogens. The relative validity of the FFQ was estimated by comparison with 7 day (d estimated diet records (EDR, n 64 and its reproducibility was evaluated by repeated administrations 6 weeks apart (n 79. Although the questionnaire underestimated significantly all intakes compared to the 7 d EDR, it had a good ranking ability (r 0.47-0.94; weighted κ 0.25-0.66 and it could reliably distinguish extreme intakes for all the estimated nutrients, except for saturated fatty acids. Furthermore, the correlation between repeated administrations was high (r 0.71-0.87 with a maximal misclassification of 7% (weighted κ 0.33-0.80. In conclusion, these results compare favourably with those reported by others and indicate that the FFQ is a satisfactorily reliable and valid instrument for ranking individuals within this study population.

  17. Determinação simultânea de teobromina, teofilina e cafeína em chás por cromatografia líquida de alta eficiência Simultaneous determination of theobromine, theophylline and caffeine in teas by high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Adriana Barreto Alves

    2002-06-01

    Full Text Available Para a realização deste trabalho, foram analisadas 10 amostras de diferentes tipos e marcas de chás com o objetivo de se quantificar teobromina, teofilina e cafeína simultaneamente. Para tanto, otimizou-se técnica de cromatografia líquida de alta eficiência (CLAE baseada na ISO 10095 (1992, utilizando-se coluna Inertsil ODS-3 (150x4 mm, 5 mm, fase móvel de ácido acético 1% + acetonitrila (95:5, v/v, fluxo de 1 mL/min e detector de UV/VIS ajustado em 273 nm. Os resultados de cafeína obtidos por esse método foram comparados com os obtidos por um método espectrofotométrico de acordo com Schormüller (1970. Não houve diferença significativa nos resultados de cafeína nas amostras de chá preto obtidos pelos dois métodos. As amostras de chá preto foram as que apresentaram maiores teores de teobromina e cafeína e nenhuma das amostras apresentou quantidades significativas de teofilina.To carry out this study, 10 samples of different kinds and brands of teas were analyzed with the purpose of quantifying simultaneously theobromine, theophylline and caffeine. For this, high performance liquid chromatography (HPLC was used based on ISO 10095 (1992. The conditions were: a reversed phase column (Inertisil ODS-3, 150x4 mm, 5 mm; acetic acid 1% + acetonitrile (95:5, v/v as mobile phase; flow of 1 ml/min and UV-VIS detector set at 273 nm. The results of caffeine obtained by this method were compared with those using a spectrophotometric method according to Schormüller (1970. In the case of black tea, no difference was observed in the caffeine, by both methods. The samples of black tea had the highest amounts of theobromine and caffeine and no sample had a significant amount of theophylline.

  18. Simultaneous determination of theobromine, theophylline and caffeine in teas by high performance liquid chromatography

    OpenAIRE

    Adriana Barreto Alves; Neura Bragagnolo

    2002-01-01

    Para a realização deste trabalho, foram analisadas 10 amostras de diferentes tipos e marcas de chás com o objetivo de se quantificar teobromina, teofilina e cafeína simultaneamente. Para tanto, otimizou-se técnica de cromatografia líquida de alta eficiência (CLAE) baseada na ISO 10095 (1992), utilizando-se coluna Inertsil ODS-3 (150x4 mm, 5 mm), fase móvel de ácido acético 1% + acetonitrila (95:5, v/v), fluxo de 1 mL/min e detector de UV/VIS ajustado em 273 nm. Os resultados de cafeína obtido...

  19. Microbial transformation of hydroxy metabolites of 1-oxohexyl derivatives of theobromine by Cunninghamella echinulata NRRL 1384.

    Science.gov (United States)

    Pekala, E; Kochan, M; Carnell, A J

    2009-01-01

    The biotransformation of pentoxifylline (PTX), propentofylline (PPT) and their racemic hydroxy metabolites ((+/-)-OHPTX and (+/-)-OHPPT) by using the fungus Cunninghamella echinulata NRRL 1384. A fungus Cunninghamella echinulata NRRL 1384 was used to catalyse the (S)-selective oxidation of the racemic hydroxy metabolites: (+/-)-OHPTX and (+/-)-OHPPT and for reduction of PTX and PPT. The first oxidation step appears to be selective and relatively fast while the second reduction step is slower and more selective with PTX. Modifications involving supplementing the bioconversion with glucose give yields and enantiomeric excess (ee) values similar to those obtained without glucose. The bioconversion of (+/-)-OHPTX gave an (R)-enantiomer (LSF-lisofylline) with a higher enantiopurity (maximum approximately 93% ee) compared to the bioconversion of (+/-)-OHPPT, when the maximum ee value for (R)-OHPPT was recorded at 83%. The conversion of (+/-)-OHPTX and (+/-)-OHPPT using Cunninghamella echinulata can be recognized as a process, which may be recommended as an alternative to the methods used to obtain (R)-OHPTX and (R)-OHPPT.

  20. Population pharmacokinetics of caffeine and its metabolites theobromine, paraxanthine and theophylline after inhalation in combination with diacetylmorphine

    NARCIS (Netherlands)

    Zandvliet, Anthe S.; Huitema, Alwin D. R.; de Jonge, Milly E.; den Hoed, Rob; Sparidans, Rolf W.; Hendriks, Vincent M.; van den Brink, Wim; van Ree, Jan M.; Beijnen, Jos H.

    2005-01-01

    The stimulant effect of caffeine, as an additive in diacetylmorphine preparations for study purposes, may interfere with the pharmacodynamic effects of diacetylmorphine. In order to obtain insight into the pharmacology of caffeine after inhalation in heroin users, the pharmacokinetics of caffeine

  1. Structure-affinity relationship in the interactions of human organic anion transporter 1 with caffeine, theophylline, theobromine and their metabolites.

    Science.gov (United States)

    Sugawara, Mitsuru; Mochizuki, Takahiro; Takekuma, Yoh; Miyazaki, Katsumi

    2005-08-15

    It is well known that human organic anion transporter 1 (hOAT1) transports many kinds of drugs, endogenous compounds, and toxins. However, little is known about the structure-affinity relationship. The aim of this study was to elucidate the structure-affinity relationship using a series of structurally related compounds that interact with hOAT1. Inhibitory effects of xanthine- and uric acid-related compounds on the transport of p-aminohippuric acid were examined using CHO-K1 cells stably expressing hOAT1. The order of potency for the inhibitory effects of xanthine-related compounds on PAH uptake was 1-methyl derivative>7-methyl derivative>3-methyl derivative falling dotsxanthine>1,3,7-trimethyl derivative (caffeine). The order of potency of the inhibition was 1,3,7-trimethyluric acid>1,3-dimethyluric acid>1,7-dimethyluric acid>1-methyluric acid>uric acid. A significant correlation between inhibitory potency and lipophilicity of the tested uric acid-related compounds was observed. The main determinant of the affinity of xanthine-related compounds is the position of the methyl group. On the other hand, lipophilicity is the main determinant of the affinity of uric acid-related compounds.

  2. Theobromine for the treatment of persistent cough: a randomised, multicentre, double-blind, placebo-controlled clinical trial.

    Science.gov (United States)

    Morice, Alyn H; McGarvey, Lorcan; Pavord, Ian D; Higgins, Bernard; Chung, Kian Fan; Birring, Surinder S

    2017-07-01

    To investigate the effect of BC1036 on health-related quality of life (QOL) in subjects with persistent cough. The secondary objective was to investigate the effect of BC1036 on subjective cough severity. This was a randomised, multicentre, double-blind, placebo-controlled, parallel-group study in 289 subjects with persistent cough. Subjects received BC1036 or placebo twice daily for 14 days. The primary endpoint comprised cough-related QOL assessed using the validated Leicester Cough Questionnaire (LCQ) at Day 14. Secondary endpoints comprised the LCQ scores at Day 7 and Day 28, cough severity VAS scores at each visit and pulmonary function tests. At baseline, mean total LCQ score in the BC1036 group was lower (i.e., worse QOL) than placebo (P<0.001), indicating significant between-group heterogeneity. Mean baseline-adjusted change in LCQ score at Day 14 was greater for BC1036 [mean (SD) 2.4±3.5] compared to placebo [mean (SD) score 2.2±3.0], but did not reach statistical significance (P=0.60). Mean cough severity VAS score decreased to a greater extent in the BC1036 group compared to placebo, but again the results were not statistically significant (-12.2±23.28 in BC1036 group and -11.0±21.34 in placebo group at Day 14, P=0.688). There was no significant change in pulmonary function measurements. The adverse event (AE) profile was similar in both groups. This study showed that BC1036 was well tolerated and, although the primary endpoint did not achieve statistical significance, the magnitude of improvement was greater with BC1036 compared to placebo with respect to improving QOL and reducing cough severity. ClinicalTrials.gov: NCT01656668.

  3. Influence of Theobromine on Heat Production and Body Temperatures in Cold-Exposed Humans: A Preliminary Report

    Science.gov (United States)

    1989-11-01

    sponsoring the research and development Include the address.) * DCIEM 9a. PROJECT OR GRANT NO. (if appropriate, the applicable research 9b. CONTRACT NO. (if...appropriate, the applicable number under and development project or grant number under which the document which the document was written) was written...ABSTRACT ( a brief and factua sunway of the documenL it may also Woper elsewhere in the body of t document itself. It is hgldy demsirbte that the abstract

  4. Synthesis, physical-chemical and biological properties of 1.8-disubstituted of theobromine. V. 8-Benzylidenhydrazino-1-p-methylbenzyltheobromines

    Directory of Open Access Journals (Sweden)

    D. G. Ivanchenko

    2015-10-01

    Full Text Available The problem of searching biologically active compounds amidst xanthine derivatives is a crucial one and is an issue for long-term investigation. Aim. In order to find new biologically active compounds among xanthine derivatives, undescribed earlier 8-benzylidenhydrazino-1-p-methylbenzyltheobromines have been synthesized. Methods and results. Reaction of 1-p-methylbenzyl-8-bromoxanthine with the excess of hydrazine hydrate in the aqueous dioxane is implemented through formation of 8-hydrazine-1(4-methylbenzyltheobromine. Through the interaction of 8-hydrazinetheobromine with aldehydes in aqueous propan-2-ol respective 8-benzylidenhydrazino-1-p-methylbenzyltheobromines have been obtained. Structure of synthesized compounds has been definitely proved by NMR-spectroscopy. Conclusions. Molecular and pharmacological descriptors of obtained substances have been calculated. The antioxidant activity of the obtained compounds has been explored. Priorities for further search of biologically active compounds in a range of xanthine derivatives have been set out.

  5. Nutrition, Metabolic Disorders and Lifestyle of Aircrew

    Science.gov (United States)

    1993-01-01

    reported 3. CAFFEINE IN THE DIET alertness. The duration and magnitude of thewe effects on Caffeine (1,3,7-trimethylxanthine), theobromine (3,7...cocoa. Theobromine is found in significant protein and carbohydrate have been administered to both quantities in cocoa but it is not believed to be

  6. Onderzoek naar de kwaliteit van chocoladerepen

    NARCIS (Netherlands)

    Sledsens, A.M.J.; Herstel, H.

    1989-01-01

    Om de kwaliteit van in willekeurige winkels gekochte chocoladerepen te kunnen beoordelen heeft Konsumenten Kontakt het RIKILT verzocht 24 monsters chocoladerepen te onderzoeken. Alle monsters zijn onderzocht op nettogewicht, totaal vet, vocht, saccharose, lactose, theobromine,

  7. Environ: E00771 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available E00771 Guarana Medicinal herb Caffeine [CPD:C07481], Theophylline [CPD:C07130], The...obromine [CPD:C07480], Tannin, Saponin Paullinia cupana [TAX:392747] Sapindaceae Guarana seed Major component: Caffeine

  8. The Contribution of cocoa additive to cigarette smoking addiction

    NARCIS (Netherlands)

    Rambali B; Andel I van; Schenk E; Wolterink G; Werken G van de; Stevenson H; Vleeming W; TOX; SIR; LVM; PZO

    2003-01-01

    In this report the effect of these compounds on the addiction to cigarette smoking was assessed, using currently available information in the literature on psychoactive compounds of cocoa. The investigated psychoactive cocoa compounds were theobromine, caffeine, serotonin, histamine, tryptophan,

  9. Theacrine, a special purine alkaloid with sedative and hypnotic properties from Cammelia assamica var. kucha in mice.

    Science.gov (United States)

    Xu, Jie-Kun; Kurihara, Hiroshi; Zhao, Liang; Yao, Xin-Sheng

    2007-01-01

    The central nervous system activities of theacrine (1,3,7,9-tetramethyluric acid), a purine alkaloid which is abundantly present in Camellia assamica var. kucha, were investigated in ambulatory activity, pentobarbital-induced sleep and forced swimming test in mice, compared with two other purine alkaloids, caffeine and theobromine. Caffeine treatment led to a marked increase in the ambulatory activity accompanied with decreasing of the immobility time in forced swimming test at both 10 and 30 mg/kg. Under the same conditions, neither theacrine nor theobromine showed obvious excited efficacy. Both doses of theacrine could significantly prolong the sleeping time induced by pentobarbital, while caffeine and theobromine exhibited an inverted effect. These results indicated that theacrine possessed potent sedative and hypnotic properties and its central nervous system effects were different from those of caffeine and theobromine.

  10. 微生物の新しい生産機能の開発と工業的応用

    OpenAIRE

    山田, 秀明; HIDEAKI, YAMADA; 関西大学工学部; Department of Biotechnology, Faculty of Engineering, Kansai University

    1993-01-01

    We have carried out studies on the industrial production of various biologically and chemically useful compounds-amino acids, amides, acids, pyrogallol, theobromine, pantothenic acid, coenzymes and polyunsaturated fatty acids-using novel productive functions of microorganisms.

  11. Extraction of purine alkaloids from maté (Ilex paraguariensis) using supercritical CO(2).

    Science.gov (United States)

    Saldaña, M D; Mohamed, R S; Baer, M G; Mazzafera, P

    1999-09-01

    Experimental data for the supercritical CO(2) extraction of purine alkaloids (caffeine, theobromine, and theophylline) from ground herbal maté tea (Ilex paraguaryensis) using a high-pressure apparatus are presented. Caffeine, theophylline, and theobromine were identified in the extracted fractions using HPLC. Results indicated a much higher CO(2) selectivity for caffeine in comparison with those for theophylline and theobromine. Solubilities of pure compounds in carbon dioxide were also determined at 313.2, 323.2, 338.2, and 343.2 K, and pressures ranging from 14 to 24 MPa. Caffeine solubility exhibited a retrograde behavior with temperature while theophylline and theobromine manifested a normal behavior at conditions explored in this study. Solubilities in binary CO(2)/purine alkaloid model systems were much higher than those obtained during extraction of maté tea, demonstrating the difficulty of using binary data in predicting complex multicomponent behavior.

  12. Pentoxifylline

    Science.gov (United States)

    ... to caffeine-containing products (coffee, tea, colas), pentoxifylline, theobromine, theophylline (Theo-Dur), or any other drugs.tell ... of reach of children. Store it at room temperature and away from excess heat and moisture (not ...

  13. Relation of 24-hour urinary caffeine and caffeine metabolite excretions with self-reported consumption of coffee and other caffeinated beverages in the general population

    National Research Council Canada - National Science Library

    Petrovic, Dusan; Estoppey Younes, Sandrine; Pruijm, Menno; Ponte, Belén; Ackermann, Daniel; Ehret, Georg; Ansermot, Nicolas; Mohaupt, Markus; Paccaud, Fred; Vogt, Bruno; Pechère-Bertschi, Antoinette; Martin, Pierre-Yves; Burnier, Michel; Eap, Chin B; Bochud, Murielle; Guessous, Idris

    2016-01-01

    ...% of caffeine is provided by caffeinated coffee consumption, and then metabolized by the liver CYP1A2 enzyme into paraxanthine (~80%), theobromine (~12%) and theophylline (~4%) [4]. Caffeine and caffeine rel...

  14. ' §"§?§§4§§é§§§'§§;§;§§a§R§;§§§§§égfigsgéééé §»2 ...

    African Journals Online (AJOL)

    temperature and relative humiciity of28 is 2 °C and. 46% respectively. Food and water were ... 21.9 2 5.8 for 'éhe high dose theobromine lreotanmt. The megm values for red blood cell countlx 106 ... and P<{l.95 for WBC) ilxerease relative to control. ' Table I : Eflect of Theobromine on Haematological Parameters ir: Rats.

  15. Characterisation of AC1: a naturally decaffeinated coffee

    Directory of Open Access Journals (Sweden)

    Luciana Benjamim Benatti

    2012-01-01

    Full Text Available We compared the biochemical characteristics of the beans of a naturally decaffeinated Arabica coffee (AC1 discovered in 2004 with those of the widely grown Brazilian Arabica cultivar "Mundo Novo" (MN. Although we observed differences during fruit development, the contents of amino acids, organic acids, chlorogenic acids, soluble sugars and trigonelline were similar in the ripe fruits of AC1 and MN. AC1 beans accumulated theobromine, and caffeine was almost entirely absent. Tests on the supply of [2-14C] adenine and enzymatic analysis of theobromine synthase and caffeine synthase in the endosperm of AC1 confirmed that, as in the leaves, caffeine synthesis is blocked during the methylation of theobromine to caffeine. The quality of the final coffee beverage obtained from AC1 was similar to that of MN.

  16. Type I photosensitized reactions of oxopurines. Kinetics and thermodynamics of the reaction with triplet benzophenone by time-resolved photoacoustic spectroscopy

    Science.gov (United States)

    Murgida, Daniel H.; Erra Balsells, Rosa; Crippa, Pier Raimondo; Viappiani, Cristiano

    1998-09-01

    Benzophenone photosensitized reactions of caffeine, theophylline and theobromine were investigated in acetonitrile by time-resolved laser-induced photoacoustics. In the three cases global quenching rate constants of triplet benzophenone were measured as a function of temperature and it was observed that this is a non-activated process. Besides, for theobromine and theophylline heats for NH hydrogen abstraction reactions were determined. In agreement with semiempirical calculation predictions, hydrogen abstraction is thermodynamically more favorable and faster for theophylline (Δ H=-265 kJ mol -1, kr=9.6×10 8 M -1 s -1) than for theobromine (Δ H=-168 kJ mol -1, kr=3.7×10 8 M -1 s -1).

  17. Some Heteroaromatic Organomercurials, Their Syntheses and Reactions: A Review of Our Research (1980-2000

    Directory of Open Access Journals (Sweden)

    Piotr Wroczynski

    2001-11-01

    Full Text Available This review reports some novel (or improved synthetic methods for preparing a number of aromatic (carbocyclic and predominantly heterocyclic organomercurials, particularly those derived from theophylline, theobromine and uracil, as well as some novel halo- and cyano-demercuration reactions. We have also synthesized the first stable organic derivative of mercury(I, viz. 1,8-bis(acetoxydimercurio theobromine, and studied its novel reactions. We have also improved the old Willgerodt method (1897, applicable for preparing various diaryliodonium chlorides from appropriate (dichloroiodoarenes and symmetric aromatic mercurials. A full list of our works, published over the past twenty years (1980-2000, is also provided (see Refs. 1-16.

  18. Standard addition strip for quantitative electrostatic spray ionization mass spectrometry analysis: determination of caffeine in drinks.

    Science.gov (United States)

    Tobolkina, Elena; Qiao, Liang; Roussel, Christophe; Girault, Hubert H

    2014-12-01

    Standard addition strips were prepared for the quantitative determination of caffeine in different beverages by electrostatic spray ionization mass spectrometry (ESTASI-MS). The gist of this approach is to dry spots of caffeine solutions with different concentrations on a polymer strip, then to deposit a drop of sample mixed with an internal standard, here theobromine on each spot and to measure the mass spectrometry signals of caffeine and theobromine by ESTASI-MS. This strip approach is very convenient and provides quantitative analyses as accurate as the classical standard addition method by MS or liquid chromatography. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Caffeine Content of Tea and Coffee

    African Journals Online (AJOL)

    1974-03-13

    Mar 13, 1974 ... The xanthines (caffeine, theophylline, and theobromine) occur in plants widely distributed throughout the world. Best known for the preparation of beverages are coffee beans which contain caffeine, tea leaves which contain caffeine and theophylline, and cocoa seeds which contain caffeine and ...

  20. Spectrometric mixture analysis: An unexpected wrinkle

    Indian Academy of Sciences (India)

    The spectrometric analysis of a mixture of two chemically and spectroscopically similar compounds is illustrated for the simultaneous spectrometric determination of caffeine and theobromine, the primary stimulants in coffee and tea, based on their ultraviolet absorbances. Their analysis indicates that such measurements ...

  1. Environ: E00818 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available E00818 Paraguay tea Medicinal herb Caffeine [CPD:C07481], Theobromine [CPD:C07480]...018], Iron [CPD:C00023], Calcium [CPD:C00076], Potassium [CPD:C00238] [DR:D08403] Ilex paraguariensis [TAX:185542] ... Aquifoliaceae Paraguay tea leaves ...

  2. Global Journal of Pure and Applied Sciences - Vol 14, No 3 (2008)

    African Journals Online (AJOL)

    Comparative effects of theobromine and Cocoa Bean Shell (CBS) extract on the performance, serum constituent profile and physiological parameters in rabbits ... of the chemical composition, the physicochemical properties of the oil extract and the amino acid profiles of the seeds of Telfairia occidentalis (fluted pumpkin) ...

  3. Author Details

    African Journals Online (AJOL)

    EBONG, P E. Vol 6, No 1 (2000) - Articles Theobromine administration inhibits platelet aggregation and elevates serum HDL - cholesterol in hyperlipidemic wistar rats. Details. ISSN: 1118-0579. AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for Librarians · for Authors · FAQ's · More about AJOL ...

  4. Author Details

    African Journals Online (AJOL)

    Comparative effects of theobromine and Cocoa Bean Shell (CBS) extract on the performance, serum constituent profile and physiological parameters in rabbits. Abstract · Vol 15, No 2 (2009) - Articles Effect Of Dietary Hydrochloric Acid (HCl) supplementation On Performance And Hysiological Response In Cockerels

  5. Discovery and Innovation - Vol 18 (2006)

    African Journals Online (AJOL)

    Theobromine rich cocoa powder induces weight loss and changes in lipid profile of obese wistar rats · EMAIL FULL TEXT EMAIL FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. MU Eteng, HA Ibekwe, UI Umoh, PE Ebong, IB Umoh, EU Eyong, 191-196. http://dx.doi.org/10.4314/dai.v18i3.15745 ...

  6. Author Details

    African Journals Online (AJOL)

    Olatunde, OA. Vol 14, No 3 (2008) - Articles Comparative effects of theobromine and Cocoa Bean Shell (CBS) extract on the performance, serum constituent profile and physiological parameters in rabbits. Abstract. ISSN: 1118-0579. AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for Librarians ...

  7. Global Journal of Pure and Applied Sciences - Vol 6, No 1 (2000)

    African Journals Online (AJOL)

    Theobromine administration inhibits platelet aggregation and elevates serum HDL - cholesterol in hyperlipidemic wistar rats. EMAIL FULL TEXT EMAIL FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. M U ETENG, E U EYONG, P E EBONG, R R ETTARH, E DAVID-OKU, I B UMOH, 89-93.

  8. A Novel Caffeine Dehydrogenase in Pseudomonas sp. Strain CBB1 Oxidizes Caffeine to Trimethyluric Acid▿

    Science.gov (United States)

    Yu, Chi Li; Kale, Yogesh; Gopishetty, Sridhar; Louie, Tai Man; Subramanian, Mani

    2008-01-01

    A unique heterotrimeric caffeine dehydrogenase was purified from Pseudomonas sp. strain CBB1. This enzyme oxidized caffeine to trimethyluric acid stoichiometrically and hydrolytically, without producing hydrogen peroxide. The enzyme was not NAD(P)+ dependent; coenzyme Q0 was the preferred electron acceptor. The enzyme was specific for caffeine and theobromine and showed no activity with xanthine. PMID:17981969

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

    Directory of Open Access Journals (Sweden)

    Euclides Lara Cardozo Junior

    2009-01-01

    Full Text Available Mate (Ilex paraguariensis St. Hil contains phytochemical compounds capable of preventing a number of healthproblems. Knowledge on the genetic contribution to the variability in these compounds can help to obtain mate progenies withhigher levels thereof in breeding programs. The composition of triterpene saponins, methylxanthines, chlorogenic acid andthe antioxidant activity of eight mate progenies were evaluated. Significant differences among progenies were verified incontents of triterpene saponins (0.003-0.080%, caffeine (0.226-1.377%, theobromine (0.176-0.831%, and chlorogenicacid (1.344-2.031% and in antioxidant activity (31.251-51.406%. The contents of theobromine were found to be negativelycorrelated with saponins and caffeine, and caffeine with chlorogenic acid, while theobromine was positively correlated withchlorogenic 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.

  10. Drug release from non-aqueous suspensions. II. The release of methylxanthines from paraffin suspensions

    NARCIS (Netherlands)

    Blaey, C.J. de; Fokkens, J.G.

    1984-01-01

    The release of 3 methylxanthines, i.e. caffeine, theobromine and theophylline, from suspensions in liquid paraffin to an aqueous phase was determined in an in vitro apparatus. The release rates were determined as a function of the pH of the aqueous phase. It was proved that the release process was

  11. Molecular and biochemical characterization of caffeine synthase and purine alkaloid concentration in guarana fruit.

    Science.gov (United States)

    Schimpl, Flávia Camila; Kiyota, Eduardo; Mayer, Juliana Lischka Sampaio; Gonçalves, José Francisco de Carvalho; da Silva, José Ferreira; Mazzafera, Paulo

    2014-09-01

    Guarana seeds have the highest caffeine concentration among plants accumulating purine alkaloids, but in contrast with coffee and tea, practically nothing is known about caffeine metabolism in this Amazonian plant. In this study, the levels of purine alkaloids in tissues of five guarana cultivars were determined. Theobromine was the main alkaloid that accumulated in leaves, stems, inflorescences and pericarps of fruit, while caffeine accumulated in the seeds and reached levels from 3.3% to 5.8%. In all tissues analysed, the alkaloid concentration, whether theobromine or caffeine, was higher in young/immature tissues, then decreasing with plant development/maturation. Caffeine synthase activity was highest in seeds of immature fruit. A nucleotide sequence (PcCS) was assembled with sequences retrieved from the EST database REALGENE using sequences of caffeine synthase from coffee and tea, whose expression was also highest in seeds from immature fruit. The PcCS has 1083bp and the protein sequence has greater similarity and identity with the caffeine synthase from cocoa (BTS1) and tea (TCS1). A recombinant PcCS allowed functional characterization of the enzyme as a bifunctional CS, able to catalyse the methylation of 7-methylxanthine to theobromine (3,7-dimethylxanthine), and theobromine to caffeine (1,3,7-trimethylxanthine), respectively. Among several substrates tested, PcCS showed higher affinity for theobromine, differing from all other caffeine synthases described so far, which have higher affinity for paraxanthine. When compared to previous knowledge on the protein structure of coffee caffeine synthase, the unique substrate affinity of PcCS is probably explained by the amino acid residues found in the active site of the predicted protein. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Caffeine biosynthesis and adenine metabolism in transgenic Coffea canephora plants with reduced expression of N-methyltransferase genes.

    Science.gov (United States)

    Ashihara, Hiroshi; Zheng, Xin-Qiang; Katahira, Riko; Morimoto, Masayuki; Ogita, Shinjiro; Sano, Hiroshi

    2006-05-01

    In anti-sense and RNA interference transgenic plants of Coffea canephora in which the expression of CaMXMT1 was suppressed, caffeine biosynthesis from [8-(14)C]adenine was investigated, together with the overall metabolism of [8-(14)C]adenine. Compared with wild type control plants, total purine alkaloid biosynthesis from adenine and conversion of theobromine to caffeine were both reduced in the transgenic plants. As found previously, [8-(14)C]adenine was metabolised to salvage products (nucleotides and RNA), to degradation products (ureides and CO(2)) and to purine alkaloids (theobromine and caffeine). In the transgenic plants, metabolism of [8-(14)C]adenine shifted from purine alkaloid synthesis to purine catabolism or salvage for nucleotides. HPLC analysis revealed a significantly reduced caffeine content in the transgenic plants. A small quantity (less than 20 nmol g(-1) fresh weight) of xanthosine had accumulated in at least one of the transgenic plants.

  13. Bioactive compounds in Mexican genotypes of cocoa cotyledon and husk.

    Science.gov (United States)

    Hernández-Hernández, Carolina; Viera-Alcaide, Isabel; Morales-Sillero, Ana María; Fernández-Bolaños, Juan; Rodríguez-Gutiérrez, Guillermo

    2018-02-01

    A characterization of the phenolic profile of 25 cocoa genotypes established in a Mexican gene bank was carried out. From five different extraction methods commonly used for phenols, extraction with acidified methanol-water was chosen as the best to quantify the concentrations of theobromine and individual phenols in cocoa beans. High concentrations of individual and total phenols were found for genotypes native to Mexico (like RIM105, M031, and M033) or from Peru and Ecuador (INI10), but not the commercial mix (CAF), and were directly associated with their antioxidant activities. Despite the loss of some theobromine and phenols during fermentation, epicatechin remained in the fermented cotyledon in high concentrations. This study could help promote the commercialization of Mexican genotypes of cocoa and reports the possibility of upcycling fermented cocoa husks, which are rich in bioactive compounds and fiber, as novel functional extracts for use in food formulation or for nutraceutical purposes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Influência do método de extração nos teores de metilxantinas em erva-mate (Ilex paraguariensis a. St.-Hil., aquifoliaceae Influence of the extraction methodology on the methylxanthines content of maté (Ilex paraguariensis a. St.-Hil., aquifoliaceae

    Directory of Open Access Journals (Sweden)

    Simone C. B. Gnoatto

    2007-04-01

    Full Text Available Ilex paraguariensis A. St. -Hil. is a native species of southern South America. The caffeine content in Ilex paraguariensis leaves has been considered an important quality parameter for maté-derived products. In this work different extraction methods for the determination of these methylxantines are compared. The influence of the extraction conditions on the methylxanthine yields was evaluated. Extraction by decoction with acidic aqueous solution (H2SO4 4N presented the higher efficiency in the theobromine extraction. The extraction in a Soxhlet with acidic aqueous solution and decoction with acidic aqueous solution showed the highest caffeine yield. For the concomitant theobromine and caffeine quantification, the decoction with acidic aqueous solution is suggested.

  15. Cocoa phytochemicals: recent advances in molecular mechanisms on health.

    Science.gov (United States)

    Kim, Jiyoung; Kim, Jaekyoon; Shim, Jaesung; Lee, Chang Yong; Lee, Ki Won; Lee, Hyong Joo

    2014-01-01

    Recent reports on cocoa are appealing in that a food commonly consumed for pure pleasure might also bring tangible benefits for human health. Cocoa consumption is correlated with reduced health risks of cardiovascular diseases, hypertension, atherosclerosis, and cancer, and the health-promoting effects of cocoa are mediated by cocoa-driven phytochemicals. Cocoa is rich in procyanidins, theobromine, (-)-epicatechin, catechins, and caffeine. Among the phytochemicals present in consumed cocoa, theobromine is most available in human plasma, followed by caffeine, (-)-epicatechin, catechin, and procyanidins. It has been reported that cocoa phytochemicals specifically modulate or interact with specific molecular targets linked to the pathogenesis of chronic human diseases, including cardiovascular diseases, cancer, neurodegenerative diseases, obesity, diabetes, and skin aging. This review summarizes comprehensive recent findings on the beneficial actions of cocoa-driven phytochemicals in molecular mechanisms of human health.

  16. Application of Surfactant Micellar Solutions as Extractants and Mobile Phases for TLC-Determination of Purine Bases and Doping Agents in Biological Liquids

    Directory of Open Access Journals (Sweden)

    Daria Victorovna Yedamenko

    2015-04-01

    Full Text Available Separation of caffeine and its metabolites (theophylline and theobromine and doping agents (spironolactone, propranolol, and ephedrine and determination of caffeine in serum sample and propranolol and ephedrine in urine were studied on normal-phase thin layers (“Sorbfil-UV-254”. Aqueous organic solvents and aqueous micellar surfactant solutions were compared as the mobile phases for separation. The acceptable separation of purine bases and doping agents was achieved by micellar Thin Layer Chromatography and normal-phase Thin Layer Chromatography. Anionic surfactant solution with added 1-propanol was the best eluent as for caffeine, theophylline, and theobromine separation, as for doping agents. The best characteristics of caffeine extraction from serum, and propranolol and ephedrine from urine were achieved when micellar eluent based on non-ionic Tween-80 surfactant was used. DOI: http://dx.doi.org/10.17807/orbital.v7i1.632

  17. Override of the radiation-induced mitotic block in human tumour cells by methylxanthines and its relationship to the potentiation of cytotoxicity

    Energy Technology Data Exchange (ETDEWEB)

    Musk, S.R.R.; Steel, G.G. (Institute of Cancer Research, Sutton (UK). Surrey Branch)

    1990-06-01

    Caffeine, theophylline, theobromine and paraxanthine, were tested for ability to override mitotic block induced by ionizing radiation in the human bladder carcinoma cell line RT112. All were found to partially override the block, at a concentration of 1mM in the order caffeine > theophylline > theobromine = paraxanthine. At a concentration of 1 mM only caffeine was found to potentiate cell killing as well as causing block override; at higher concentrations all had a significant effect on survival but little or no further influence on the degree of block override. It is concluded that override of a mitotic block is not in itself sufficient to cause increased killing when irradiated cells are incubated in the presence of caffeine, and that caffeine exerts its potentiating effect by directly inhibiting repair of damage in DNA or by causing override of radiation-induced inhibition of DNA synthesis. (author).

  18. The comparison of biological activity of chocolates made by different technological procedures

    OpenAIRE

    Lucia Godočiková; Eva Ivanišová; Július Árvay; Jana Petrová; Miroslava Kačániová

    2016-01-01

    Chocolate is one of the most consumed delicacies in the world. Nowadays, raw chocolates without vanilla or allergens are getting more attention. The aim of this study was to evaluate and compare the biological activity of different types of chocolate - cold processed chocolate and chocolate made by traditional way. Total content of fat, crude fibre, polyphenols, flavonoids, phenolic acids and metylxantines - theobromine and caffeine was evaluated. The antioxidant activity was determined by a...

  19. Metabolism of alkaloids in coffee plants

    OpenAIRE

    ASHIHARA, Hiroshi

    2006-01-01

    Coffee beans contain two types of alkaloids, caffeine and trigonelline, as major components. This review describes the distribution and metabolism of these compounds. Caffeine is synthesised from xanthosine derived from purine nucleotides. The major biosynthetic route is xanthosine -> 7-methylxanthosine -> 7-methylxanthine -> theobromine -> caffeine. Degradation activity of caffeine in coffee plants is very low, but catabolism of theophylline is always present. Theophylline is converted to xa...

  20. Does Caffeine Consumption Modify Cerebrospinal Fluid Amyloid-β Levels in Patients with Alzheimer's Disease?

    Science.gov (United States)

    Travassos, Maria; Santana, Isabel; Baldeiras, Inês; Tsolaki, Magda; Gkatzima, Olymbia; Sermin, Genc; Yener, Görsev G; Simonsen, Anja; Hasselbalch, Steen G; Kapaki, Elisabeth; Mara, Bourbouli; Cunha, Rodrigo A; Agostinho, Paula; Blennow, Kaj; Zetterberg, Henrik; Mendes, Vera M; Manadas, Bruno; de Mendon, Alexandreça

    2015-01-01

    Caffeine may be protective against Alzheimer's disease (AD) by modulating amyloid-β (Aβ) metabolic pathways. The present work aimed to study a possible association of caffeine consumption with the cerebrospinal fluid (CSF) biomarkers, particularly Aβ. The study included 88 patients with AD or mild cognitive impairment. The consumption of caffeine and theobromine was evaluated using a validated food questionnaire. Quantification of caffeine and main active metabolites was performed with liquid chromatography coupled to tandem mass spectrometry. The levels of A(1-42), total tau, and phosphorylated tau in the CSF were determined using sandwich ELISA methods and other Aβ species, Aβ(X-38), Aβ(X-40), and Aβ(X-42), with the MSD Aβ Triplex assay. The concentration of caffeine was 0.79±1.15 μg/mL in the CSF and 1.20±1.88 μg/mL in the plasma. No correlation was found between caffeine consumption and Aβ42 in the CSF. However, a significant positive correlation was found between the concentrations of theobromine, both in the CSF and in the plasma, with Aβ42 in the CSF. Theobromine in the CSF was positively correlated with the levels of other xanthines in the CSF, but not in the plasma, suggesting that it may be formed by central metabolic pathways. In conclusion, caffeine consumption does not modify the levels of CSF biomarkers, and does not require to be controlled for when measuring CSF biomarkers in a clinical setting. Since theobromine is associated with a favorable Aβ profile in the CSF, the possibility that it might have a protective role in AD should be further investigated.

  1. Distribution coefficients of purine alkaloids in water-ammonium sulfate-alkyl acetate-dialkyl phthalate systems

    Science.gov (United States)

    Korenman, Ya. I.; Krivosheeva, O. A.; Mokshina, N. Ya.

    2012-12-01

    The distribution of purine alkaloids (caffeine, theobromine, theophylline) was studied in the systems: alkyl acetates-dialkyl phtalate-salting-out agent (ammonium sulfate). The quantitative characteristics of the extraction-distribution coefficients ( D) and the degree of extraction ( R, %) are calculated. The relationships between the distribution coefficients of alkaloids and the length of the hydrocarbon radical in the molecule of alkyl acetate (dialkyl phtalate) are determined. The possibility of predicting the distribution coefficients is demonstrated.

  2. Cambios en la características físicas y químicas de granos de cacao (Theobroma cacao) fermentados con transferencia manual y semi-mecanizada, entre las cajas de fermentación

    OpenAIRE

    Peláez, Pedro. P.; Saulo Guerra; David Contreras

    2016-01-01

    The aim of this study was to evaluate variation in the physical and chemical properties of fermented cocoa beans with cocoa beans transfer between wooden fermentation boxes manually (M) and semi-mechanized (SM) way. Mass temperature, moisture, pH, and total acidity of the cotyledon and pulp; the total polyphenol, anthocyanin, reducing sugar, theobromine, and caffeine content in fresh, fermented, and dried beans; and percentage of fermented beans and time required to move beans during fermenta...

  3. Sleep Management User’s Guide for Special Operations Personnel

    Science.gov (United States)

    1992-07-01

    as detined by body temperature . The circadian trough occurs between 0200 and 0600 of the time zone to which the body is adapted. There is a...to eight-hour period each nint. Every aspect of each individual (physical and mental performance, mood, body temperature , blood pressure, pulse, etc...caffeine, and even chocolate contains a stimulating caffeine-like drug called theobromine . Many medications also contain caffeine. A number of over-the

  4. Human Platelet Senescence Study.

    Science.gov (United States)

    1980-03-01

    pH 8.6 at room temperature at 150 volts for 2 to 3 hours. Most of the agarose is then removed and a strip of the re- maining electrophoresed...the mM range: Zn++>Cu++>Fe++>Mn-H-, This cAMP phosphodiesterase appears to be unique in that it is not inhibited by 10 mM caffeine, theobromine

  5. Supercritical carbon dioxide extraction of methylxanthines from maté tea leaves

    OpenAIRE

    Saldaña, M.D.A.; R. S. Mohamed; Mazzafera, P.

    2000-01-01

    Methylxanthines are alkaloids found in natural products such as tea, coffee and guaraná. These alkaloids are commonly used in cola drinks and pharmaceutical products due principally to their stimulant and diuretic effects on the human organism. In this work, experimental data on the supercritical CO2 extraction of caffeine, theophylline and theobromine from herbal maté tea, a beverage traditionally consumed by the gauchos of southern Brazil, the Argentine, Paraguay and Uruguay, were obtained ...

  6. Soldier Performance in Continuous Operations. Administrative Manual for a Briefing and Seminar for Command and Staff Personnel

    Science.gov (United States)

    1985-07-01

    Muscular performance capability is increased most by’caffeine when necessary. In some situations it might be appropriate to and least by theobromine . "In...in operating in feine drinks, or cola preparations can achieve the desired goal a hot climate. At daily mean temperatures of 90’F, soldiers although...show no improved work at this temperature raises the requirement to 8 quarts per results."" day, while soldiers doing heavy work for 8 hours at this

  7. Computationally-Assisted Approach to the Vibrational Spectra of Molecular Crystals: Study of Hydrogen-Bonding and Pseudo-Polymorphism13

    OpenAIRE

    Nolasco, Mariela M.; Amado, Ana M.; Ribeiro-Claro, Paulo J. A.

    2006-01-01

    A new computationally-assisted methodology (PiMM), which accounts for the effects of intermolecular interactions in the crystal, is applied to the complete assignment of the Raman and infrared vibrational spectra of room temperature forms of crystalline caffeine, theobromine, and theophylline. The vibrational shifts due to crystal packing interactions are evaluated from ab initio calculations for a set of suitable molecular pairs, using the B3LYP/6-31G* approach.The proposed methodology provi...

  8. Environ: E00818 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available E00818 Paraguay tea Medicinal herb Caffeine [CPD:C07481], Theobromine [CPD:C07480],...18], Iron [CPD:C00023], Calcium [CPD:C00076], Potassium [CPD:C00238] [DR:D08403] Ilex paragua...riensis [TAX:185542] Aquifoliaceae Paraguay tea leaves Medicinal herbs [BR:br08322] Dicot plants: asterids Aquifoliaceae (holly family) E00818 Paraguay tea ...

  9. Blood pressure and endothelial function in healthy, pregnant women after acute and daily consumption of flavanol-rich chocolate: a pilot, randomized controlled trial.

    Science.gov (United States)

    Mogollon, Jaime Andres; Bujold, Emmanuel; Lemieux, Simone; Bourdages, Mélodie; Blanchet, Claudine; Bazinet, Laurent; Couillard, Charles; Noël, Martin; Dodin, Sylvie

    2013-04-08

    Several randomized clinical trials (RCTs) indicate that flavanol-rich chocolate has beneficial effects on flow-mediated dilation (FMD) and blood pressure (BP). However, no RCTs have evaluated these outcomes in pregnant women. The objective of this 2-group, parallel, double-blind RCT was to examine the effects of flavanol-rich chocolate on FMD and BP in pregnant women with normal BP. Forty-four healthy, pregnant women were randomized to the high-flavanol (n = 23) or low-flavanol (n = 21) chocolate consumption for 12 weeks. At randomization (0, 60, 120 and 180 min after a single 40-g dose of chocolate), 6 and 12 weeks after daily 20-g chocolate intake, we evaluated plasma concentrations of flavanols and theobromine, as well as the FMD and BP. Plasma epicatechin was significantly increased (p flavanol chocolate compared to low-flavanol chocolate. Theobromine concentrations were significantly higher 180 min and 12 weeks after the intake of experimental chocolate or low-flavanol chocolate (p flavanol-rich chocolate to an equivalent placebo during pregnancy and demonstrate higher plasma epicatechin and theobromine concentration in the intervention group after acute ingestion ClinicalTrials.gov Identifier: NCT01659060.

  10. Blood pressure and endothelial function in healthy, pregnant women after acute and daily consumption of flavanol-rich chocolate: a pilot, randomized controlled trial

    Science.gov (United States)

    2013-01-01

    Background Several randomized clinical trials (RCTs) indicate that flavanol-rich chocolate has beneficial effects on flow-mediated dilation (FMD) and blood pressure (BP). However, no RCTs have evaluated these outcomes in pregnant women. The objective of this 2-group, parallel, double-blind RCT was to examine the effects of flavanol-rich chocolate on FMD and BP in pregnant women with normal BP. Methods Forty-four healthy, pregnant women were randomized to the high-flavanol (n = 23) or low-flavanol (n = 21) chocolate consumption for 12 weeks. At randomization (0, 60, 120 and 180 min after a single 40-g dose of chocolate), 6 and 12 weeks after daily 20-g chocolate intake, we evaluated plasma concentrations of flavanols and theobromine, as well as the FMD and BP. Results Plasma epicatechin was significantly increased (p chocolate compared to low-flavanol chocolate. Theobromine concentrations were significantly higher 180 min and 12 weeks after the intake of experimental chocolate or low-flavanol chocolate (p chocolate to an equivalent placebo during pregnancy and demonstrate higher plasma epicatechin and theobromine concentration in the intervention group after acute ingestion Trial registration ClinicalTrials.gov Identifier: NCT01659060 PMID:23565841

  11. Metabolic Relations between Methylxanthines and Methyluric Acids in Coffea L. 1

    Science.gov (United States)

    Petermann, Joseph B.; Baumann, Thomas W.

    1983-01-01

    Metabolism of purine alkaloids in the leaves of Coffea dewevrei De Wild et Durand var excelsa Chev, Coffea liberica Bull ex Hiern and Coffea abeokutae Cramer was studied by analyzing leaf discs collected during vegetative development and by feeding the following radioactive tracers: [14C]theobromine, [14C]caffeine, and [14C]theacrine (1,3,7,9-tetramethyluric acid). Their principal metabolites were quantitatively and qualitatively determined. All three species convert the precursors to the same radioactive products, and proceed through the same four maturity stages characterized by the alkaloid accumulation pattern and by a particular transformation potency: (stage 1) young plant accumulating caffeine, transforms theobromine to caffeine; (stage 2) caffeine is gradually replaced by theacrine, theobromine and caffeine are converted to theacrine; (stage 3) theacrine disappears whereas liberine (O(2), 1,9-thrimethyluric acid) accumulates, theacrine is metabolized to liberine; (stage 4) branched-out plant containing liberine but no theacrine, caffeine is converted rapidly to liberine via theacrine. Methylliberine (O(2),1,7,9-tetramethyluric acid), presumably the direct precursor of liberine, is occasionally found in low concentrations at stage 3 and 4. The collective term `liberio-excelsoid' introduced by geneticists for the numerous races or species of Pachycoffea is in accordance with the phytochemical equality found in this work. PMID:16663351

  12. Metabolic Relations between Methylxanthines and Methyluric Acids in Coffea L.

    Science.gov (United States)

    Petermann, J B; Baumann, T W

    1983-12-01

    Metabolism of purine alkaloids in the leaves of Coffea dewevrei De Wild et Durand var excelsa Chev, Coffea liberica Bull ex Hiern and Coffea abeokutae Cramer was studied by analyzing leaf discs collected during vegetative development and by feeding the following radioactive tracers: [(14)C]theobromine, [(14)C]caffeine, and [(14)C]theacrine (1,3,7,9-tetramethyluric acid). Their principal metabolites were quantitatively and qualitatively determined. All three species convert the precursors to the same radioactive products, and proceed through the same four maturity stages characterized by the alkaloid accumulation pattern and by a particular transformation potency: (stage 1) young plant accumulating caffeine, transforms theobromine to caffeine; (stage 2) caffeine is gradually replaced by theacrine, theobromine and caffeine are converted to theacrine; (stage 3) theacrine disappears whereas liberine (O(2), 1,9-thrimethyluric acid) accumulates, theacrine is metabolized to liberine; (stage 4) branched-out plant containing liberine but no theacrine, caffeine is converted rapidly to liberine via theacrine. Methylliberine (O(2),1,7,9-tetramethyluric acid), presumably the direct precursor of liberine, is occasionally found in low concentrations at stage 3 and 4.The collective term ;liberio-excelsoid' introduced by geneticists for the numerous races or species of Pachycoffea is in accordance with the phytochemical equality found in this work.

  13. Morphometric and functional abnormalities of kidneys in the progeny of mice fed chocolate during pregnancy and lactation.

    Directory of Open Access Journals (Sweden)

    Ewa Skopińska-Rózewska

    2006-09-01

    Full Text Available Even most commonly consumed beverages like tea, coffee, chocolate and cocoa contain methylxanthines, biogenic amines and polyphenols, among them catechins, that exhibit significant biological activity and might profoundly affect the organism homeostasis. We have previously shown that 400 mg of bitter chocolate or 6 mg of theobromine added to the daily diet of pregnant and afterwards lactating mice affected embryonic angiogenesis and caused bone mineralization disturbances as well as limb shortening in 4-weeks old offspring. The aim of the present study was the morphometric and functional evaluation of kidneys in the 4-weeks old progeny mice fed according to the protocol mentioned above. Progeny from the mice fed chocolate presented considerable morphometric abnormalities in the kidney structure, with the lower number of glomeruli per mm2 and their increased diameter. Moreover, higher serum creatinine concentration was observed in that group of offspring. No morphometric or functional irregularities were found in the progeny of mice fed theobromine. Abnormalities demonstrated in the offspring of mice fed chocolate are not related to its theobromine content. Consequently, identification of active compound(s responsible for the observed effects is of vital importance.

  14. Effect of methylxanthines on acetaminophen hepatotoxicity in various induction states.

    Science.gov (United States)

    Kalhorn, T F; Lee, C A; Slattery, J T; Nelson, S D

    1990-01-01

    The effect of caffeine, theophylline and theobromine on acetaminophen-induced hepatotoxicity was evaluated in uninduced, 3-methylcholanthrene- and phenobarbital-induced adult male Sprague-Dawley rats. The methylxanthines themselves did not cause hepatotoxicity in any induction state. In 3-methylcholanthrene-induced rats, each methylxanthine afforded protection (in varying degrees) against acetaminophen-induced hepatotoxicity as reflected by serum alanine aminotransferase and liver histopathology determined 24 hr after acetaminophen administration. However, in phenobarbital-induced rats, caffeine and theophylline substantially potentiated the hepatotoxicity of acetaminophen whereas theobromine had no effect. Hepatic glutathione (GSH) was determined in rats that received caffeine 4 hr after acetaminophen or vehicle. Acetaminophen alone substantially depleted hepatic GSH in each induction state, whereas caffeine depleted hepatic GSH in uninduced and phenobarbital-induced, but not in 3-methylcholanthrene-induced rats. In rats that received both caffeine and acetaminophen together, hepatic GSH depletion was greater than in rats that received acetaminophen only. The effect of caffeine on hepatic GSH is most likely due to a decrease in core body temperature. The most likely mechanisms for the effects observed are 1) inhibition of acetaminophen reactive metabolite formation in 3-methylcholanthrene-induced animals by each of the methylxanthines, and 2) activation of the phenobarbital-inducible forms of cytochrome(s) P-450 toward formation of acetaminophen reactive metabolites by caffeine and theophylline, but not theobromine.

  15. Genetic and Phytochemical Analysis to Evaluate the Diversity and Relationships of Mate (Ilex paraguariensis A.St.-Hil.) Elite Genetic Resources in a Germplasm Collection.

    Science.gov (United States)

    Friedrich, Juliana Cristhina; Gonela, Adriana; Gonçalves Vidigal, Maria Celeste; Vidigal Filho, Pedro Soares; Sturion, José Alfredo; Cardozo Junior, Euclides Lara

    2017-03-01

    The aim of this study was to evaluate the phytochemical and genetic diversity, relationships and identification of mate (Ilex paraguariensis A.St.-Hil.) elite genetic resources belonging to the Brazilian germplasm collection and mate breeding program. Mate has been studied due to the presence of phytochemical compounds, especially methylxanthines and phenolic compounds. The samples were collected from the leaves of 76 mate elite genetic resources (16 progenies × 5 localities). Total DNA was extracted from mate leaves and 20 random primers were used for DNA amplification. Methylxanthines (caffeine and theobromine) and phenolic compounds (chlorogenic, neochlorogenic, and criptochlorogenic acids) were quantified by HPLC. The genetic divergence estimated was higher within (92%) than among (8%) the different populations. Analysis of genetic distance between origins provided the formation of two groups by UPGMA cluster analysis, with higher polymorphism (94.9%). The average content of caffeine ranged from 0.01 to 1.38% and theobromine of 0.10 - 0.85% (w/w). The caffeoylquinic acids concentrations (1.43 - 5.38%) showed a gradient 3-CQA > 5-CQA > 4-CQA. The coefficient of genetic variation (CVg) was of low magnitude for all mono-caffeoylquinics acids. Significant correlations (positive and negative) were observed between the phytochemical compounds. Genetic diversity analysis performed by RAPD markers showed a greater intra-populational diversity; genetic resources with low caffeine and higher theobromine content were identified and can be used in breeding programs; the correlation between methylxanthines and phenolic compounds can be used as a good predictor in future studies. © 2017 Wiley-VHCA AG, Zurich, Switzerland.

  16. Flavanols and methylxanthines in commercially available dark chocolate: a study of the correlation with nonfat cocoa solids.

    Science.gov (United States)

    Langer, Swen; Marshall, Lisa J; Day, Andrea J; Morgan, Michael R A

    2011-08-10

    Intake of flavanols, a subgroup of dietary polyphenols present in many fruits and vegetables, may be associated with health benefits, particularly with reducing the risk of coronary diseases. Cocoa and chocolate products are rich in flavanol monomers, oligomers, and polymers (procyanidins). This study used normal phase HPLC to detect, identify, and quantify epicatechin, catechin, total monomers, procyanidin oligomers and polymers in 14 commercially available chocolate bars. In addition, methylxanthines (theobromine and caffeine) were also quantified. Nonfat cocoa solids (NFCS) were determined both gravimetrically and by calculation from theobromine contents. The flavanol levels of 12 commonly consumed brands of dark chocolate have been quantified and correlated with % theobromine and % NFCS. Epicatechin comprised the largest fraction of total chocolate flavonoids, with the remainder being catechin and procyanidins. Calculated NFCS did not reflect epicatechin (R(2) = 0.41) or total flavanol contents (R(2) = 0.49). Epicatechin (R(2) = 0.96) was a reliable marker of total flavanols, catechin (R(2) = 0.67) to a lesser extent. All dark chocolate tested contained higher levels of total flavanols (93.5-651.1 mg of epicatechin equiv/100 g of product) than a milk or a white "chocolate" (40.6 and 0.0 mg of epicatechin equiv/100 g, respectively). The amount and integrity of procyanidins often suffer in the manufacturing of chocolate, chiefly due to oxidation and alkalinization. In this study, the labeled cocoa content of the chocolate did not always reflect analyzed levels of flavonoids. Increasingly, high % NFCS is being used commercially to reflect chocolate quality. If the flavanol content of chocolate is accepted to be a key determinant of health benefits, then continued monitoring of flavanol levels in commercially available chocolate products may be essential for consumer assurance.

  17. Cytochrome P450-dependent metabolism of caffeine in Drosophila melanogaster.

    Science.gov (United States)

    Coelho, Alexandra; Fraichard, Stephane; Le Goff, Gaëlle; Faure, Philippe; Artur, Yves; Ferveur, Jean-François; Heydel, Jean-Marie

    2015-01-01

    Caffeine (1, 3, 7-trimethylxanthine), an alkaloid produced by plants, has antioxidant and insecticide properties that can affect metabolism and cognition. In vertebrates, the metabolites derived from caffeine have been identified, and their functions have been characterized. However, the metabolites of caffeine in insects remain unknown. Thus, using radiolabelled caffeine, we have identified some of the primary caffeine metabolites produced in the body of Drosophila melanogaster males, including theobromine, paraxanthine and theophylline. In contrast to mammals, theobromine was the predominant metabolite (paraxanthine in humans; theophylline in monkeys; 1, 3, 7-trimethyluric acid in rodents). A transcriptomic screen of Drosophila flies exposed to caffeine revealed the coordinated variation of a large set of genes that encode xenobiotic-metabolizing proteins, including several cytochromes P450s (CYPs) that were highly overexpressed. Flies treated with metyrapone--an inhibitor of CYP enzymes--showed dramatically decreased caffeine metabolism, indicating that CYPs are involved in this process. Using interference RNA genetic silencing, we measured the metabolic and transcriptomic effect of three candidate CYPs. Silencing of CYP6d5 completely abolished theobromine synthesis, whereas CYP6a8 and CYP12d1 silencing induced different consequences on metabolism and gene expression. Therefore, we characterized several metabolic products and some enzymes potentially involved in the degradation of caffeine. In conclusion, this pioneer approach to caffeine metabolism in insects opens novel perspectives for the investigation of the physiological effects of caffeine metabolites. It also indicates that caffeine could be used as a biomarker to evaluate CYP phenotypes in Drosophila and other insects.

  18. Cytochrome P450-dependent metabolism of caffeine in Drosophila melanogaster.

    Directory of Open Access Journals (Sweden)

    Alexandra Coelho

    Full Text Available Caffeine (1, 3, 7-trimethylxanthine, an alkaloid produced by plants, has antioxidant and insecticide properties that can affect metabolism and cognition. In vertebrates, the metabolites derived from caffeine have been identified, and their functions have been characterized. However, the metabolites of caffeine in insects remain unknown. Thus, using radiolabelled caffeine, we have identified some of the primary caffeine metabolites produced in the body of Drosophila melanogaster males, including theobromine, paraxanthine and theophylline. In contrast to mammals, theobromine was the predominant metabolite (paraxanthine in humans; theophylline in monkeys; 1, 3, 7-trimethyluric acid in rodents. A transcriptomic screen of Drosophila flies exposed to caffeine revealed the coordinated variation of a large set of genes that encode xenobiotic-metabolizing proteins, including several cytochromes P450s (CYPs that were highly overexpressed. Flies treated with metyrapone--an inhibitor of CYP enzymes--showed dramatically decreased caffeine metabolism, indicating that CYPs are involved in this process. Using interference RNA genetic silencing, we measured the metabolic and transcriptomic effect of three candidate CYPs. Silencing of CYP6d5 completely abolished theobromine synthesis, whereas CYP6a8 and CYP12d1 silencing induced different consequences on metabolism and gene expression. Therefore, we characterized several metabolic products and some enzymes potentially involved in the degradation of caffeine. In conclusion, this pioneer approach to caffeine metabolism in insects opens novel perspectives for the investigation of the physiological effects of caffeine metabolites. It also indicates that caffeine could be used as a biomarker to evaluate CYP phenotypes in Drosophila and other insects.

  19. Two Distinct Pathways for Metabolism of Theophylline and Caffeine Are Coexpressed in Pseudomonas putida CBB5▿ †

    Science.gov (United States)

    Yu, Chi Li; Louie, Tai Man; Summers, Ryan; Kale, Yogesh; Gopishetty, Sridhar; Subramanian, Mani

    2009-01-01

    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. PMID:19447909

  20. Ex vivo human platelet aggregation induced by decompression during reduced barometric pressure, hydrostatic, and hydrodynamic (Bernoulli) effect.

    Science.gov (United States)

    Murayama, M

    1984-03-01

    Decompression of human platelet-rich plasma (PRP) in siliconized glass or plastic to 380 mm Hg for 3 hours at 38 degrees C produced platelet aggregation independent of pO2. Aggregation also took place when PRP was compressed to 8,000 PSI and then decompressed slowly to one atmosphere (14.7 PSI) without gas bubble formation. Platelets also aggregated when plasma was decompressed hydrodynamically (Bernoulli effect) at room temperature. It was also found that the drugs piracetam (2-oxypyrolidine acetamide) and pentoxifylline (1-(5-oxohexyl)-theobromine) at 0.5 and 1.0 mM prevent thrombocyte aggregation. Implications for mountain sickness are discussed.

  1. Quantitative capillary electrophoresis and its application in analysis of alkaloids in tea, coffee, coca cola, and theophylline tablets.

    Science.gov (United States)

    Li, Mengjia; Zhou, Junyi; Gu, Xue; Wang, Yan; Huang, Xiaojing; Yan, Chao

    2009-01-01

    A quantitative CE (qCE) system with high precision has been developed, in which a 4-port nano-valve was isolated from the electric field and served as sample injector. The accurate amount of sample was introduced into the CE system with high reproducibility. Based on this system, consecutive injections and separations were performed without voltage interruption. Reproducibilities in terms of RSD lower than 0.8% for retention time and 1.7% for peak area were achieved. The effectiveness of the system was demonstrated by the quantitative analysis of caffeine, theobromine, and theophylline in real samples, such as tea leaf, roasted coffee, coca cola, and theophylline tablets.

  2. Purine and its analogues and radiation damage in Bacillus megaterium spores

    Energy Technology Data Exchange (ETDEWEB)

    Powers, E.L.

    1986-12-01

    As an extension of results obtained from radiation studies on caffeine both in other laboratories and more recently in this laboratory using the bacterial spore as the test system, six compounds with chemical structures closely resembling that of caffeine were tested as radiation modifiers. Of these compounds, purine, adenine and hypoxanthine resembled caffeine in sensitizing spores to radiation, while theobromine, xanthine and theophylline did not. These responses are discussed in relation to the electron sequestration hypothesis of cellular sensitization to high-energy radiation.

  3. The potential of methylxanthine-based therapies in pediatric respiratory tract diseases.

    Science.gov (United States)

    Oñatibia-Astibia, Ainhoa; Martínez-Pinilla, Eva; Franco, Rafael

    2016-03-01

    Caffeine, theophylline and theobromine are the most known methylxanthines as they are present in coffee, tea and/or chocolate. In the last decades, a huge experimental effort has been devoted to get insight into the variety of actions that these compounds exert in humans. From such knowledge it is known that methylxanthines have a great potential in prevention, therapy and/or management of a variety of diseases. The benefits of methylxanthine-based therapies in the apnea of prematurity and their translational potential in pediatric affections of the respiratory tract are here presented. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Methylxanthines: properties and determination in various objects

    Science.gov (United States)

    Andreeva, Elena Yu; Dmitrienko, Stanislava G.; Zolotov, Yurii A.

    2012-05-01

    Published data on the properties and determination of caffeine, theophylline, theobromine and some other methylxanthines in various objects are surveyed and described systematically. Different sample preparation procedures such as liquid extraction from solid matrices and liquid-liquid, supercritical fluid and solid-phase extraction are compared. The key methods of analysis including chromatography, electrophoresis, spectrometry and electrochemical methods are discussed. Examples of methylxanthine determination in plants, food products, energy beverages, pharmaceuticals, biological fluids and natural and waste waters are given. The bibliography includes 393 references.

  5. Sorption of methylxanthines by different sorbents

    Science.gov (United States)

    Dmitrienko, S. G.; Andreeva, E. Yu.; Tolmacheva, V. V.; Terent'eva, E. A.

    2013-05-01

    Sorption of caffeine, theophylline, theobromine, diprophylline, and pentoxyphylline on different sorbents (supercross-linked polystyrene, surface-modified copolymer of styrene and divinylbenzene Strata-X, and carbon nanomaterials Taunit and Diasorb-100-C16T) was studied in a static mode in an effort to find new sorbents suitable for sorption isolation and concentration of methylxanthines. The peculiarities of sorption of methylxanthines were explained in relation to the solution acidity, the nature of the sorbates and their concentration, the nature of the solvent, and the structural characteristics of the sorbents.

  6. Antioxidant effect of naturally occurring xanthines on the oxidative damage of DNA bases; Effet antioxydant de xanthines naturelles sur le dommage oxydant des bases de l`ADN

    Energy Technology Data Exchange (ETDEWEB)

    Vieira, A.J.S.C.; Telo, J.P.; Pereira, H.F.; Patrocinio, P.F. [Instituto Superior Tecnico, Lisbon (Portugal); Dias, R.M.B. [Instituto Tecnologico e Nuclear, Sacavem codex (Portugal). Dept. de Quimica

    1999-01-01

    The repair of the oxidised radicals of adenine and guanosine by several naturally occurring xanthines was studied. Each pair of DNA purine/xanthine was made to react with the sulphate radical and the decrease of the concentration of both compounds was measured by HPLC as a function of irradiation time. The results show that xanthine efficiently prevents the oxidation of the two DNA purines. Theophylline and para-xanthine repair the oxidizes radical of adenine but not the one from guanosine. Theobromine and caffeine to do not show any protecting effect. An order of the oxidation potentials of all the purines studied is proposed. (authors) 10 refs.

  7. Changes in physical and chemical characteristics of fermented cocoa(Theobroma cacaobeans with manual and semi-mechanized transfer, between fermentation boxes

    Directory of Open Access Journals (Sweden)

    Pedro. P. Peláez

    2016-01-01

    Full Text Available The aim of this study was to evaluate variation in the physical and chemical properties of fermented cocoa beans with cocoa beans transfer between wooden fermentation boxes manually (M and semi - mechanized (SM way. Mass temperature, moisture, pH, and total acidity of the cotyledon and pulp; the total polyphenol, anthocyanin, reducing sugar, theobromine, and caffeine content in fresh, fermented, and dried beans; and percentage of fermented beans and time required to move beans during fermentation were determined. The cocoa used grew in the Pachiza district of the San Martin region of Peru. Cocoa sampling w as each 0, 48, 72, 96, 120, 144, and 168 h of fermentation. The cocoa mass temperature was highest with both removal systems after 96 h of fermentation. M cotyledon and pulp samples had the highest moisture content and titratable acidity, while cotyledon a nd pulp pH with both systems were statistically equal. In contrast, fermented beans had a higher polyphenol, anthocyanin, reducing sugar, theobromine, and caffeine content with SM. SM produced the greatest amount of fermentation (91.67% and required the s hortest amount of time to move beans (78.56 min. In conclusion, the system of fermentation of cocoa beans with SM was faster and produced fermented grains with high chemical quality.

  8. Health Benefits of Methylxanthines in Cacao and Chocolate

    Science.gov (United States)

    Franco, Rafael; Oñatibia-Astibia, Ainhoa; Martínez-Pinilla, Eva

    2013-01-01

    One may wonder why methylxanthines are so abundant in beverages used by humans for centuries, or in cola-drinks that have been heavily consumed since their appearance. It is likely that humans have stuck to any brew containing compounds with psychoactive properties, resulting in a better daily life, i.e., more efficient thinking, exploring, hunting, etc., however, without the serious side effects of drugs of abuse. The physiological effects of methylxanthines have been known for a long time and they are mainly mediated by the so-called adenosine receptors. Caffeine and theobromine are the most abundant methylxanthines in cacao and their physiological effects are notable. Their health-promoting benefits are so remarkable that chocolate is explored as a functional food. The consequences of adenosine receptor blockade by natural compounds present in cacao/chocolate are here reviewed. Palatability and health benefits of methylxanthines, in general, and theobromine, in particular, have further contributed to sustain one of the most innocuous and pleasant habits: chocolate consumption. PMID:24145871

  9. Effects of high flavanol dark chocolate on cardiovascular function and platelet aggregation.

    Science.gov (United States)

    Rull, Gurvinder; Mohd-Zain, Zetty N; Shiel, Julian; Lundberg, Martina H; Collier, David J; Johnston, Atholl; Warner, Timothy D; Corder, Roger

    2015-08-01

    Regular consumption of chocolate and cocoa products has been linked to reduced cardiovascular mortality. This study compared the effects of high flavanol dark chocolate (HFDC; 1064mg flavanols/day for 6weeks) and low flavanol dark chocolate (LFDC; 88mg flavanols/day for 6weeks) on blood pressure, heart rate, vascular function and platelet aggregation in men with pre-hypertension or mild hypertension. Vascular function was assessed by pulse wave analysis using radial artery applanation tonometry in combination with inhaled salbutamol (0.4mg) to assess changes due to endothelium-dependent vasodilatation. HFDC did not significantly reduce blood pressure compared to baseline or LFDC. Heart rate was increased by LFDC compared to baseline, but not by HFDC. Vascular responses to salbutamol tended to be greater after HFDC. Platelet aggregation induced by collagen or the thromboxane analogue U46619 was unchanged after LFDC or HFDC, whereas both chocolates reduced responses to ADP and the thrombin receptor activator peptide, SFLLRNamide (TRAP6), relative to baseline. Pre-incubation of platelets with theobromine also attenuated platelet aggregation induced by ADP or TRAP6. We conclude that consumption of HFDC confers modest improvements in cardiovascular function. Platelet aggregation is modulated by a flavanol-independent mechanism that is likely due to theobromine. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. The Effect of Caffeine Ingestion during Evening Exercise on Subsequent Sleep Quality in Females.

    Science.gov (United States)

    Ali, A; O'Donnell, J M; Starck, C; Rutherfurd-Markwick, K J

    2015-06-01

    In a randomised, double-blind, placebo-controlled crossover design, 10 females taking monophasic oral contraceptives completed 90 min intermittent treadmill-running 45 min after ingestion of 6 mg∙kg(-1) body mass anhydrous caffeine or artificial sweetener (placebo). Water (3 mL∙kg(-1)) was provided every 15 min during exercise. Venous blood samples were taken before, during and after exercise, as well as after sleep (~15 h post-ingestion), and levels of caffeine, paraxanthine, theobromine and theophylline were measured using high-performance liquid chromatography. Sleep quality was assessed using the Leeds Sleep Evaluation Questionnaire. Plasma caffeine concentration peaked 100 min after ingestion. Caffeine clearance was 0.95±0.14 mL·min(-1)·kg(-1) while the elimination half-life of caffeine was 17.63±8.06 h. Paraxanthine and theophylline levels were significantly elevated at 15 h with no significant change in theobromine. Sleep latency and subsequent quality of sleep was impaired following caffeine supplementation (Psleep quality in female athletes taking a low-dose monophasic oral contraceptive steroid following an intermittent-exercise running protocol. The data shows that female athletes using monophasic oral contraceptive steroids will have impaired sleep quality following evening caffeine ingestion. © Georg Thieme Verlag KG Stuttgart · New York.

  11. Degradation of exogenous caffeine by Populus alba and its effects on endogenous caffeine metabolism.

    Science.gov (United States)

    Pierattini, Erika C; Francini, Alessandra; Raffaelli, Andrea; Sebastiani, Luca

    2016-04-01

    This is the first study reporting the presence of endogenous caffeine, theobromine, and theophylline in all organs of poplar plants. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was used in order to evaluate the uptake, translocation, and metabolism of caffeine-(trimethyl-(13)C) in Populus alba L. Villafranca clone grown in hydroponic conditions. We investigated the remediation of caffeine since it is one of the most widely consumed drugs and it is frequently detected in wastewater treatment plant effluents, surface water, and groundwater worldwide. Our results demonstrated that poplar can absorb and degrade exogenous caffeine without negative effects on plant health. Data showed that concentrations of all endogenous compounds varied depending on caffeine-(trimethyl-(13)C) treatments. In particular, in control conditions, endogenous caffeine, theobromine, and theophylline were mainly distributed in roots. On the other hand, once caffeine-(trimethyl-(13)C) was provided, this compound and its dimethy-(13)C metabolites are mainly localized at leaf level. In conclusion, our results support the possible use of Villafranca clone in association with other water treatment systems in order to complete the process of caffeine remediation.

  12. Forensic Analysis of Stains on Fabric Using Direct Analysis in Real-time Ionization with High-Resolution Accurate Mass-Mass Spectrometry.

    Science.gov (United States)

    Kern, Sara E; Crowe, John B; Litzau, Jonathan J; Heitkemper, Douglas T

    2017-06-12

    A rapid technique using direct analysis in real-time (DART) ambient ionization coupled to a high-resolution accurate mass-mass spectrometer (HRAM-MS) was employed to analyze stains on an individual's pants suspected to have been involved in a violent crime. The victim was consuming chocolate ice cream at the time of the attack, and investigators recovered the suspect's pants exhibiting splatter stains. Liquid chromatography with mass spectral detection (LC-MS) and stereoscopic light microscopy (SLM) were also utilized in this analysis. It was determined that the stains on the pants contained theobromine and caffeine, known components of chocolate. A shard from the ceramic bowl that contained the victim's ice cream and a control chocolate ice cream sample were also found to contain caffeine and theobromine. The use of DART-HRAM-MS was useful in this case due to its rapid analysis capability and because of the limited amount of sample present as a stain. Published 2017. This article is a U.S. Government work and is in the public domain in the USA.

  13. Health Benefits of Methylxanthines in Cacao and Chocolate

    Directory of Open Access Journals (Sweden)

    Rafael Franco

    2013-10-01

    Full Text Available One may wonder why methylxanthines are so abundant in beverages used by humans for centuries, or in cola-drinks that have been heavily consumed since their appearance. It is likely that humans have stuck to any brew containing compounds with psychoactive properties, resulting in a better daily life, i.e., more efficient thinking, exploring, hunting, etc., however, without the serious side effects of drugs of abuse. The physiological effects of methylxanthines have been known for a long time and they are mainly mediated by the so-called adenosine receptors. Caffeine and theobromine are the most abundant methylxanthines in cacao and their physiological effects are notable. Their health-promoting benefits are so remarkable that chocolate is explored as a functional food. The consequences of adenosine receptor blockade by natural compounds present in cacao/chocolate are here reviewed. Palatability and health benefits of methylxanthines, in general, and theobromine, in particular, have further contributed to sustain one of the most innocuous and pleasant habits: chocolate consumption.

  14. Health benefits of methylxanthines in cacao and chocolate.

    Science.gov (United States)

    Franco, Rafael; Oñatibia-Astibia, Ainhoa; Martínez-Pinilla, Eva

    2013-10-18

    One may wonder why methylxanthines are so abundant in beverages used by humans for centuries, or in cola-drinks that have been heavily consumed since their appearance. It is likely that humans have stuck to any brew containing compounds with psychoactive properties, resulting in a better daily life, i.e., more efficient thinking, exploring, hunting, etc., however, without the serious side effects of drugs of abuse. The physiological effects of methylxanthines have been known for a long time and they are mainly mediated by the so-called adenosine receptors. Caffeine and theobromine are the most abundant methylxanthines in cacao and their physiological effects are notable. Their health-promoting benefits are so remarkable that chocolate is explored as a functional food. The consequences of adenosine receptor blockade by natural compounds present in cacao/chocolate are here reviewed. Palatability and health benefits of methylxanthines, in general, and theobromine, in particular, have further contributed to sustain one of the most innocuous and pleasant habits: chocolate consumption.

  15. Polyphenol-Retaining Decaffeinated Cocoa Powder Obtained by Supercritical Carbon Dioxide Extraction and Its Antioxidant Activity

    Directory of Open Access Journals (Sweden)

    Kinji Kobori

    2013-10-01

    Full Text Available Cocoa beans contain many functional ingredients such as theobromine and polyphenols, but also contain a relatively high amount of caffeine, which can negatively impact human health. It is therefore desirable to reduce caffeine levels in cocoa powder used to make chocolate or cocoa beverages while retaining functional ingredients. We have established conditions for supercritical carbon dioxide (SCCO2 extraction that remove 80.1% of the caffeine from cocoa powder while retaining theobromine (94.1% and polyphenols (84.7%. The antioxidant activity of the decaffeinated cocoa powder (DCP made with this optimized SCCO2 extraction method was 85.3% that of non-processed cocoa powder. The total procyanidin and total polyphenol concentrations of the DCPs resulting from various SCCO2 extractions showed a significant positive correlation with oxygen radical absorbance capacity (ORAC. The correlation coefficient between total polyphenols and ORAC was higher than that between total procyanidins and ORAC; thus, the concentration of total polyphenols might be a greater factor in the antioxidant activity of DCP. These results indicate that we could remove large quantities of caffeine from conventional high-cocoa products while retaining the functional benefits of high polyphenol content. This SCCO2 extraction method is expected to be applicable high-cocoa products, such as dark chocolate.

  16. Changes in physical and chemical characteristics of fermented cocoa (Theobroma cacao beans with manual and semi-mechanized transfer, between fermentation boxes

    Directory of Open Access Journals (Sweden)

    Pedro. P. Peláez

    2016-06-01

    Full Text Available The aim of this study was to evaluate variation in the physical and chemical properties of fermented cocoa beans with cocoa beans transfer between wooden fermentation boxes manually (M and semi-mechanized (SM way. Mass temperature, moisture, pH, and total acidity of the cotyledon and pulp; the total polyphenol, anthocyanin, reducing sugar, theobromine, and caffeine content in fresh, fermented, and dried beans; and percentage of fermented beans and time required to move beans during fermentation were determined. The cocoa used grew in the Pachiza district of the San Martin region of Peru. Cocoa sampling was each 0, 48, 72, 96, 120, 144, and 168 h of fermentation. The cocoa mass temperature was highest with both removal systems after 96 h of fermentation. M cotyledon and pulp samples had the highest moisture content and titratable acidity, while cotyledon and pulp pH with both systems were statistically equal. In contrast, fermented beans had a higher polyphenol, anthocyanin, reducing sugar, theobromine, and caffeine content with SM. SM produced the greatest amount of fermentation (91.67% and required the shortest amount of time to move beans (78.56 min. In conclusion, the system of fermentation of cocoa beans with SM was faster and produced fermented grains with high chemical quality.

  17. Supercritical carbon dioxide extraction of methylxanthines from maté tea leaves

    Directory of Open Access Journals (Sweden)

    M.D.A. Saldaña

    2000-09-01

    Full Text Available Methylxanthines are alkaloids found in natural products such as tea, coffee and guaraná. These alkaloids are commonly used in cola drinks and pharmaceutical products due principally to their stimulant and diuretic effects on the human organism. In this work, experimental data on the supercritical CO2 extraction of caffeine, theophylline and theobromine from herbal maté tea, a beverage traditionally consumed by the gauchos of southern Brazil, the Argentine, Paraguay and Uruguay, were obtained using high pressure extraction equipment that allows adequate control of temperature and pressure. The continuous extraction/fractionation of maté tea leaves, Ilex paraguariensis in natura using carbon dioxide was carried out at 313.2 and 343.2 K and pressures of 13.8 and 25.5 MPa. Extraction/fractionation curves revealed the large influence of temperature and pressure on extraction yield. CO2 was also found to show a higher selectivity for caffeine than for theophylline and theobromine.

  18. Caffeine markedly sensitizes human mesothelioma cell lines to pemetrexed

    Science.gov (United States)

    Min, Sang Hee; Goldman, I. David; Zhao, Rongbao

    2013-01-01

    Pemetrexed is a new generation antifolate approved for the treatment of mesothelioma and non-small cell lung cancer. Caffeine is known to augment radiation or chemotherapeutic drug-induced cell killing. The current study addresses the impact of caffeine on the activity of pemetrexed in mesothelioma cell lines. Caffeine enhanced pemetrexed activity in all four mesothelioma cell lines tested (H2052, H2373, H28 and MSTO-211H). Caffeine sensitized H2052 cells in a dose- and schedule-dependent manner, and was associated with a markedly decreased clonogenic survival. Caffeine sensitization occurred only in cells subjected to pulse, but not continuous, exposure to pemetrexed. Similar pemetrexed sensitization was also observed with the clinically better tolerated caffeine analog, theobromine. Pemetrexed sensitization by caffeine was associated with an increase in pemetrexed-induced phosphorylation of ataxia-telangiectasia-mutated (ATM) and Chk1. These data indicate that caffeine and its analog, theobromine, may be a useful approach to enhance pemetrexed-based chemotherapy. PMID:17594092

  19. DETECTION AND QUANTIFICATION OF PHYTOCHEMICAL MARKERS OF Ilex paraguariensis BY LIQUID CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    Rodrigo M. C. Pinto

    2015-11-01

    Full Text Available Ilex paraguariensis (yerba-mate is used as a beverage, and its extract requires adequate quality control methods in order to guarantee quality and safe use. Strategies to develop and optimize a chromatographic method to quantify theobromine, caffeine, and chlorogenic acid in I. paraguariensis extracts were evaluated by applying a quality by design (QbD model and ultra high-performance liquid chromatography (UHPLC. The presence of these three phytochemical markers in the extracts was evaluated using UHPLC-MS and was confirmed by the chromatographic bands in the total ion current traces (m/z of 181.1 [M+H]+, 195.0 [M+H]+, and 353.0 [M−H]−, respectively. The developed method was then transferred to a high-performance liquid chromatography (HPLC platform, and the three phytochemical markers were used as external standards in the validation of a method for analyses of these compounds in extracts using a diode array detector (DAD. The validated method was applied to quantify the chlorogenic acid, caffeine, and theobromine in the samples. HPLC-DAD chromatographic fingerprinting was also used in a multivariate approach to process the entire data and to separate the I. paraguariensis extracts into two groups. The developed method is very useful for qualifying and quantifying I. paraguariensis extracts.

  20. [Louis-Marie Rousseau and the "Chocolat rationnel des pharmaciens français" (Rational Chocolate of French pharmacists)].

    Science.gov (United States)

    Raynal, Cécile

    2015-03-01

    In 1883, the chemist Louis-Marie Rousseau (1849-1930) creats the "Compagnie hygiénique française" (French Hygienic Company). The company manufactures and sells the "Poudre de viande Rousseau" (Rousseau meat powder) and the "Chocolat Rousseau" (Rousseau Chocolate) by methods developed and patented by the pharmacist. Ten years after a successful collaboration, L.-M. Rousseau separates from his associates and founds the "Chocolaterie spéciale d'Ermont" (Special Chocolate factory of Ermont) in the village of Ermont near Paris. Here is manufactured the "Chocolat Rationnel des pharmaciens français" (Rational Chocolate of French pharmacists), hygienic chocolate sold only in pharmacies. The factory is also a pharmaceutical laboratory where is extracted theobromine from waste vegetable substances of cocoa. It then produces the "Théobromine Rousseau cristallisée" (crystallized Rousseau's Theobromine) sold as tablets, then the "Théosol" that will be commercialized until the middle of 1930s.

  1. The cardiovascular benefits of dark chocolate.

    Science.gov (United States)

    Kerimi, Asimina; Williamson, Gary

    2015-08-01

    Dark chocolate contains many biologically active components, such as catechins, procyanidins and theobromine from cocoa, together with added sucrose and lipids. All of these can directly or indirectly affect the cardiovascular system by multiple mechanisms. Intervention studies on healthy and metabolically-dysfunctional volunteers have suggested that cocoa improves blood pressure, platelet aggregation and endothelial function. The effect of chocolate is more convoluted since the sucrose and lipid may transiently and negatively impact on endothelial function, partly through insulin signalling and nitric oxide bioavailability. However, few studies have attempted to dissect out the role of the individual components and have not explored their possible interactions. For intervention studies, the situation is complex since suitable placebos are often not available, and some benefits may only be observed in individuals showing mild metabolic dysfunction. For chocolate, the effects of some of the components, such as sugar and epicatechin on FMD, may oppose each other, or alternatively in some cases may act together, such as theobromine and epicatechin. Although clearly cocoa provides some cardiovascular benefits according to many human intervention studies, the exact components, their interactions and molecular mechanisms are still under debate. Copyright © 2015 Elsevier Inc. All rights reserved.

  2. Simultaneous quantification of caffeine and its three primary metabolites in rat plasma by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Choi, Eu Jin; Bae, Soo Hyeon; Park, Jung Bae; Kwon, Min Jo; Jang, Su Min; Zheng, Yu Fen; Lee, Young Sun; Lee, Su-Jun; Bae, Soo Kyung

    2013-12-01

    A rapid, sensitive, simple and accurate LC-MS/MS method for the simultaneous quantitation of caffeine, and its three primary metabolites, theobromine, paraxanthine, and theophylline, in rat plasma was developed and validated. Chromatographic separation was performed on an Agilent Poroshell 120 EC-C18 column using 1 μg/mL acetaminophen as an internal standard. Each sample was run at 0.5 mL/min for a total run time of 7 min/sample. Detection and quantification were performed using a mass spectrometer in selected reaction-monitoring mode with positive electrospray ionization. The lower limit of quantification was 5 ng/mL for all analytes with linear ranges up to 5000 ng/mL for caffeine and 1000 ng/mL for its metabolites. The coefficient of variation for assay precision was less than 12.6%, with an accuracy of 93.5-114%. The assay was successfully applied to determine plasma concentrations of caffeine, theobromine, paraxanthine, and theophylline in rat administered various energy drinks containing the same caffeine content. Various energy drinks exhibited considerable variability in the pharmacokinetic profiles of caffeine and its three primary metabolites, even containing the same caffeine. Different additives of energy drinks might contribute to these results. Copyright © 2013 Elsevier Ltd. All rights reserved.

  3. Associations of ambulatory blood pressure with urinary caffeine and caffeine metabolite excretions.

    Science.gov (United States)

    Guessous, Idris; Pruijm, Menno; Ponte, Belén; Ackermann, Daniel; Ehret, Georg; Ansermot, Nicolas; Vuistiner, Philippe; Staessen, Jan; Gu, Yumei; Paccaud, Fred; Mohaupt, Markus; Vogt, Bruno; Pechère-Bertschi, Antoinette; Pechère-Berstchi, Antoinette; Martin, Pierre-Yves; Burnier, Michel; Eap, Chin B; Bochud, Murielle

    2015-03-01

    Intake of caffeinated beverages might be associated with reduced cardiovascular mortality possibly via the lowering of blood pressure. We estimated the association of ambulatory blood pressure with urinary caffeine and caffeine metabolites in a population-based sample. Families were randomly selected from the general population of Swiss cities. Ambulatory blood pressure monitoring was conducted using validated devices. Urinary caffeine, paraxanthine, theophylline, and theobromine excretions were measured in 24 hours urine using ultrahigh performance liquid chromatography tandem mass spectrometry. We used mixed models to explore the associations of urinary excretions with blood pressure although adjusting for major confounders. The 836 participants (48.9% men) included in this analysis had mean age of 47.8 and mean 24-hour systolic and diastolic blood pressure of 120.1 and 78.0 mm Hg. For each doubling of caffeine excretion, 24-hour and night-time systolic blood pressure decreased by 0.642 and 1.107 mm Hg (both P values theobromine excretion was not associated with blood pressure. Anti-hypertensive therapy, diabetes mellitus, and alcohol consumption modify the association of caffeine urinary excretion with systolic blood pressure. Ambulatory systolic blood pressure was inversely associated with urinary excretions of caffeine and other caffeine metabolites. Our results are compatible with a potential protective effect of caffeine on blood pressure. © 2014 American Heart Association, Inc.

  4. Increases in cerebrospinal fluid caffeine concentration are associated with favorable outcome after severe traumatic brain injury in humans

    Science.gov (United States)

    Sachse, Kathleen T; Jackson, Edwin K; Wisniewski, Stephen R; Gillespie, Delbert G; Puccio, Ava M; Clark, Robert SB; Dixon, C Edward; Kochanek, Patrick M

    2013-01-01

    Caffeine, the most widely consumed psychoactive drug and a weak adenosine receptor antagonist, can be neuroprotective or neurotoxic depending on the experimental model or neurologic disorder. However, its contribution to pathophysiology and outcome in traumatic brain injury (TBI) in humans is undefined. We assessed serial cerebrospinal fluid (CSF) concentrations of caffeine and its metabolites (theobromine, paraxanthine, and theophylline) by high-pressure liquid chromatography/ultraviolet in 97 ventricular CSF samples from an established bank, from 30 adults with severe TBI. We prospectively selected a threshold caffeine level of ≥1 μmol/L (194 ng/mL) as clinically significant. Demographics, Glasgow Coma Scale (GCS) score, admission blood alcohol level, and 6-month dichotomized Glasgow Outcome Scale (GOS) score were assessed. Mean time from injury to initial CSF sampling was 10.77±3.13 h. On initial sampling, caffeine was detected in 24 of 30 patients, and the threshold was achieved in 9 patients. Favorable GOS was seen more often in patients with CSF caffeine concentration ≥ versus theobromine and paraxanthine were also associated with favorable outcome (P = 0.018 and 0.056, respectively). Caffeine and its metabolites are commonly detected in CSF in patients with severe TBI and in an exploratory assessment are associated with favorable outcome. We speculate that caffeine may be neuroprotective by long-term upregulation of adenosine A1 receptors or acute inhibition of A2a receptors. PMID:17684518

  5. Cloning, expression, crystallization and preliminary X-ray analysis of the XMT and DXMT N-methyltransferases from Coffea canephora (robusta)

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, Andrew A., E-mail: andrewmc@embl.fr [European Molecular Biology Laboratory, 6 Rue Jules Horowitz, BP 181, 38042 Grenoble (France); Biget, Laurent [Nestlé Research and Development, 101 Avenue Gustave Eiffel, Notre-Dame D’Oe, 37097 Tours (France); Lin, Chenwei [Department of Plant Breeding and Genetics, Department of Plant Biology, Cornell University, Ithaca, NY 14853 (United States); Petiard, Vincent [Nestlé Research and Development, 101 Avenue Gustave Eiffel, Notre-Dame D’Oe, 37097 Tours (France); Tanksley, Steve D. [Department of Plant Breeding and Genetics, Department of Plant Biology, Cornell University, Ithaca, NY 14853 (United States); McCarthy, James G. [Nestlé Research and Development, 101 Avenue Gustave Eiffel, Notre-Dame D’Oe, 37097 Tours (France); European Molecular Biology Laboratory, 6 Rue Jules Horowitz, BP 181, 38042 Grenoble (France)

    2007-04-01

    The genes encoding XMT and DXMT, the enzymes from Coffea canephora (robusta) that catalyse the three independent N-methyl transfer reactions in the caffeine-biosynthesis pathway, have been cloned and the proteins have been expressed in Escherichia coli. Both proteins have been crystallized in the presence of the demethylated cofactor S-adenosyl-l-cysteine (SAH) and substrate (xanthosine for XMT and theobromine for DXMT). Caffeine is a secondary metabolite produced by a variety of plants including Coffea canephora (robusta) and there is growing evidence that caffeine is part of a chemical defence strategy protecting young leaves and seeds from potential predators. The genes encoding XMT and DXMT, the enzymes from Coffea canephora (robusta) that catalyse the three independent N-methyl transfer reactions in the caffeine-biosynthesis pathway, have been cloned and the proteins have been expressed in Escherichia coli. Both proteins have been crystallized in the presence of the demethylated cofactor S-adenosyl-l-cysteine (SAH) and substrate (xanthosine for XMT and theobromine for DXMT). The crystals are orthorhombic, with space group P2{sub 1}2{sub 1}2{sub 1} for XMT and C222{sub 1} for DXMT. X-ray diffraction to 2.8 Å for XMT and to 2.5 Å for DXMT have been collected on beamline ID23-1 at the ESRF.

  6. Study on inclusion complex of cyclodextrin with methyl xanthine derivatives by fluorimetry

    Science.gov (United States)

    Wei, Yan-Li; Ding, Li-Hua; Dong, Chuan; Niu, Wei-Ping; Shuang, Shao-Min

    2003-10-01

    The inclusion complexes of β-cyclodextrin (β-CD) and HP-β-cyclodextrin (HP-β-CD) with caffeine, theophylline and theobromine were investigated by fluorimetry. Various factors affecting the formation of inclusion complexes were discussed in detail including forming time, pH effect and temperature. The results indicate that inclusion process was affected seriously by laying time and pH. The forming time of β-CD inclusion complexes is much longer than that of HP-β-CD. The optimum pH range is about 7-12 for caffeine, 8-10 for TP, 10.5-12 for TB. The intensities of their fluorescence increase with the decreasing of temperature. Their maximum excitation wavelengths are all in the range of 280-290 nm. The emission wavelength of caffeine and theophylline are both in the range of 340-360 nm, and that of theobromine is about 325 nm. The fluorescence signals are intensified with the increasing concentration of CD. The stoichiometry of the inclusion complexes of CD with these three methyl xanthine derivatives are all 1:1 and the formation constant are all calculated.

  7. Application of near-infrared reflectance spectroscopy to the simultaneous prediction of alkaloids and phenolic substances in green tea leaves.

    Science.gov (United States)

    Schulz, H; Engelhardt, U H; Wegent, A; Drews, H; Lapczynski, S

    1999-12-01

    A near-infrared reflectance spectroscopic (NIRS) method for the prediction of polyphenol and alkaloid compounds in the leaves of green tea [Camellia sinensis (L.) O. Kuntze] was developed. Reference measurements of the individual catechins, gallic acid, caffeine, and theobromine were performed by reversed-phase HPLC. The total polyphenols were determined according to the colorimetric Folin-Ciocalteu assay. Using the partial least-squares algorithm, very good calibration statistics were obtained for the prediction of gallic acid, (-)-epicatechin, (-)-epigallocatechin, (-)-epicatechin gallate, (-)-epigallocatechin gallate, caffeine, and theobromine (R(2) > 0.85) with standard deviation/standard error of cross-validation (SD/SECV) ratio ranging from 2.00 to 6.27. Simultaneously, the dry matter content of the tea leaves can be analyzed very precisely (R(2) = 0.94; SD/SECV = 4.12). Furthermore, it is possible to discriminate tea leaves of different age by principal component analysis on the basis of the received NIR spectra. Prediction of the total polyphenol content is performed with a lower accuracy, which might be due to the lack of specificity in the colorimetric reference method. The study demonstrates that NIRS technology can be successfully applied as a rapid method not only for breeding and cultivation purposes but also to estimate the quality and taste of green tea and to control industrial processes, for example, decaffeination.

  8. Analysis of xanthines in beverages using a fully automated SPE-SPC-DAD hyphenated system

    Energy Technology Data Exchange (ETDEWEB)

    Medvedovici, A. [Bucarest Univ., Bucarest (Romania). Faculty of Chemistry, Dept. of Analytical Chemistry; David, F.; David, V.; Sandra, P. [Research Institute of Chromatography, Kortrijk (Belgium)

    2000-08-01

    Analysis of some xanthines (caffeine, theophylline and theobromine) in beverages has been achieved by a fully automated on-line Solid Phase Extraction - Supercritical Fluid Chromatography - Diode Array Detection (Spe - Sofc - Dad). Three adsorbents have been tested for the Spe procedure: octadecyl modified silicagel (ODS) and two types of styrene-divinylbenzen copolymer based materials, from which Porapack proved to be the most suitable adsorbent. Optimisation and correlation of both Spe and Sofc operational parameters are also discussed. By this technique, caffeine was determined in ice tea and Coca-Cola in a concentration of 0.15 ppm, theobromine - 1.5 ppb, and theophylline - 0.15 ppb. [Italian] Si e' realizzata l'analis di alcune xantine (caffeina, teofillina e teobromina) mediante un sistema, in linea, completamente automatizzato basato su Estrazione in Fase Solida - Cromatografia in Fase Supercritica - Rivelazione con Diode Array (Spe - Sfc - Dad). Per la procedura Spe sono stati valutati tre substrati: silice ottadecilica (ODS) e due tipi di materiali polimerici a base stirene-divinilbenzene, di cui, quello denominato PRP-1, e' risultato essere il piu' efficiente. Sono discusse sia l'ottimizzazione che la correlazione dei parametri operazionali per la Spe e la Sfc. Con questa tecnica sono state determinate, in te' ghiacciato e Coca-Cola, la caffeina, la teobromina e la teofillina alle concentrazini di 0.15, 1.5 e 0.15 ppm.

  9. A rapid simultaneous determination of methylxanthines and proanthocyanidins in Brazilian guaraná (Paullinia cupana Kunth.).

    Science.gov (United States)

    Machado, Kamilla Nunes; Freitas, Aline Alves de; Cunha, Luzia Helena; Faraco, André Augusto Gomes; Pádua, Rodrigo Maia de; Braga, Fernão Castro; Vianna-Soares, Cristina Duarte; Castilho, Rachel Oliveira

    2018-01-15

    Paullinia cupana is a plant native to Brazil that is widely used in traditional medicine as a physical and mental stimulant. It is also used worldwide to produce soft drinks. A method for the simultaneous quantitation of seven markers in guaraná by HPLC-PDA was developed, and extraction methods for the determination of methylxanthines and tannins were investigated. Quantified substances were theobromine, theophylline, caffeine, catechin, epicatechin, procyanidins A2 and B2. Results confirmed the satisfactory selectivity and linearity (r2≥0.99) within the mass ranges. Repeatability (RSD≤2.80%), intermediate precision (RSD≤4.47%), accuracy (recoveries from 90.59%-104.67%), and robustness were demonstrated. Extract 1 presented the contents: 0.0177% (±1.02%) for theobromine, 0.0131% (±1.14%) for theophylline, 2.9429% (±1.27%) for caffeine, 0.4563% (±1.02%) for catechin, 0.5515% (±1.05%) for epicatechin, 0.0607% (±2.80%) for A2 and 0.1035% (±1.39%) for B2. The method for simultaneous quantitation of seven chemical markers in guaraná proved to be reliable using a simple and convenient HPLC setup. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Polyethylene Glycols as Efficient Catalysts for the Oxidation of Xanthine Alkaloids by Ceric Ammonium Nitrate in Acetonitrile: A Kinetic and Mechanistic Approach

    Directory of Open Access Journals (Sweden)

    S. Shylaja

    2013-01-01

    Full Text Available Kinetics of oxidation of xanthine alkaloids, such as Xanthine (XAN, hypoxanthine (HXAN, caffeine (CAF, theophylline (TPL, and theobromine (TBR, have been studied with ceric ammonium nitrate (CAN using poly ethylene glycols (PEG as catalysts. Reaction obeyed first order kinetics in both [CAN] and [Xanthine alkaloid]. Highly sluggish CAN-xanthine alkaloid reactions (in acetonitrile media even at elevated temperatures are enhanced in presence PEGs (PEG-200, -300, -400, -600. An increase in [PEG] increased the rate of oxidation linearly. This observation coupled with a change in absorption of CAN in presence of PEG, [H–(OCH2–CH2n–O–NH4Ce(NO34(CH3CN] (PEG bound CAN species, is considered to be more reactive than CAN. The mechanism of oxidation in PEG media has been explained by Menger-Portnoy’s enzymatic model.

  11. Physicochemical properties and antioxidant capacity of raw, roasted and puffed cacao beans.

    Science.gov (United States)

    Hu, SuJung; Kim, Byung-Yong; Baik, Moo-Yeol

    2016-03-01

    The antioxidant capacity and attributable bioactive compounds of puffed cacao beans were investigated. Roasting was carried out at 190°C for 15min and puffing was performed at 4-7kgf/cm(2). Cacao beans puffed at 4kgf/cm(2) showed the highest total polyphenols (23.16mgGAE/gsample) and total flavonoids (10.65mgCE/gsample) (pcacao beans reflected the total polyphenols and flavonoids measured. The quantities of theobromine, catechin, epicatechin, and procyanidin B2 were higher in cacao beans puffed at 4kgf/cm(2) than in roasted cacao beans. Puffed cacao beans received a good sensory score in flavor, but sourness increased as puffing pressure increased. Thus, these results suggest that, in cacao bean processing, puffing could be an alternative to roasting, which provide a rich taste and high antioxidant capacity. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. A TLC visualisation reagent for dimethylamphetamine and other abused tertiary amines.

    Science.gov (United States)

    Kato, N; Ogamo, A

    2001-01-01

    Application of citric acid/acetic anhydride reagent (CAR), a colour reagent selective for tertiary amines in solution, improves detection of abused tertiary amino drugs on the TLC plate. The plate is pretreated by a brief immersion in phosphoric acid/acetone solution to suppress colouration. After suppressing, the plate is sprayed with CAR and heated at 100 degrees C, causing tertiary amines to turn red purple within 3 minutes. The sensitivity of this new CAR method is 2.5 to 15-times greater than that of conventional detection with Dragendorff reagent for some of the tertiary amines dimethylamphetamine, methylephedrine, levomepromazine, chlorpromazine, caffeine, theophylline, theobromine and nicotine. This present method provides rapid TLC detection of abused tertiary amino drugs such as phenethylamine, phenothiazine, xanthine derivative, nicotine and narcotics.

  13. Cloning, expression, crystallization and preliminary X-ray analysis of the XMT and DXMT N-methyltransferases from Coffea canephora (robusta).

    Science.gov (United States)

    McCarthy, Andrew A; Biget, Laurent; Lin, Chenwei; Petiard, Vincent; Tanksley, Steve D; McCarthy, James G

    2007-04-01

    Caffeine is a secondary metabolite produced by a variety of plants including Coffea canephora (robusta) and there is growing evidence that caffeine is part of a chemical defence strategy protecting young leaves and seeds from potential predators. The genes encoding XMT and DXMT, the enzymes from Coffea canephora (robusta) that catalyse the three independent N-methyl transfer reactions in the caffeine-biosynthesis pathway, have been cloned and the proteins have been expressed in Escherichia coli. Both proteins have been crystallized in the presence of the demethylated cofactor S-adenosyl-L-cysteine (SAH) and substrate (xanthosine for XMT and theobromine for DXMT). The crystals are orthorhombic, with space group P2(1)2(1)2(1) for XMT and C222(1) for DXMT. X-ray diffraction to 2.8 A for XMT and to 2.5 A for DXMT have been collected on beamline ID23-1 at the ESRF.

  14. [Determination of six main components in compound theophylline tablet by convolution curve method after prior separation by column partition chromatography

    Science.gov (United States)

    Zhang, S. Y.; Wang, G. F.; Wu, Y. T.; Baldwin, K. M. (Principal Investigator)

    1993-01-01

    On a partition chromatographic column in which the support is Kieselguhr and the stationary phase is sulfuric acid solution (2 mol/L), three components of compound theophylline tablet were simultaneously eluted by chloroform and three other components were simultaneously eluted by ammonia-saturated chloroform. The two mixtures were determined by computer-aided convolution curve method separately. The corresponding average recovery and relative standard deviation of the six components were as follows: 101.6, 1.46% for caffeine; 99.7, 0.10% for phenacetin; 100.9, 1.31% for phenobarbitone; 100.2, 0.81% for theophylline; 99.9, 0.81% for theobromine and 100.8, 0.48% for aminopyrine.

  15. EFFECT OF NATURALLY OCCURRING XANTHINES ON BACTERIA. I. ANTIMICROBIAL ACTION AND POTENTIATING EFFECT ON ANTIBIOTIC SPECTRA.

    Science.gov (United States)

    RAJ, C V; DHALA, S

    1965-05-01

    The effect of xanthines on various microorganisms was studied. The antibacterial effect was not high; most of the test organisms could easily withstand a concentration of 2,500 mug/ml. Caffeine was more antibacterial than theophylline, and the latter more than theobromine. Caffeine citrate exhibited greater inhibitory effect than did pure caffeine. The effect was both bacteriostatic and bactericidal against susceptible organisms. The susceptibility of organisms to xanthines differed greatly even in related species. The morphology of Aerobacter aerogenes and A. cloacae was affected under the influence of caffeine; filamentation of cells followed sublethal doses. Potentiation was seen with antibiotics and caffeine; resistant strains were killed with a lower dose of drug in the presence of caffeine. This potentiating effect was pronounced with the tetracyclines; with streptomycin, the effect was the contrary.

  16. The use of ultrasound in the extraction of Ilex paraguariensis leaves: a comparison with maceration.

    Science.gov (United States)

    Jacques, Rosângela Assis; dos Santos Freitas, Lisiane; Pérez, Valéria Flores; Dariva, Cláudio; de Oliveira, Ana Paula; de Oliveira, José Vladimir; Caramão, Elina Bastos

    2007-01-01

    The objective of this work is to discuss the main parameters that influence the sonication extraction of Ilex paraguariensis leaves. For this purpose, the extraction time, solvent polarity, solvent volume, sample mass and particle size were evaluated. Results showed that the main variable affecting the extraction process was the solvent polarity. Though in a less extent, temperature and extraction time also demonstrated to be important parameters, while the other variables did not present a significant influence on the extraction yield. The extracts at the optimized condition were compared with those obtained by maceration, in terms of mass yield and chemical composition. The major compounds identified in the extracts were caffeine and palmitic acid. Some saturated hydrocarbons such as fatty acids, fatty acid methyl esters, phytosterols, and theobromine were also identified in the fractions.

  17. The measurement of theophylline in human serum or plasma using gas chromatography and isotope dilution-mass spectrometry (GC-IDMS) taking other substituted xanthines into consideration.

    Science.gov (United States)

    Kress, Michael; Meissner, Dieane; Kaiser, Patricia; Hanke, Rainer; Wood, William Graham

    2002-01-01

    A method is described which uses a combination of gas chromatography and isotope dilution-mass spectrometry (GC-IDMS) to determine the concentration of theophylline (1,3-dimethyl xanthine) in human plasma or serum samples. The effects of similar substituted xanthines - namely theobromine (3,7-dimethyl xanthine), paraxanthine (1,7-dimethyl xanthine) 1,3-dimethyl-7-(2-hydroxyethyl) xanthine (internal standard HPLC) and caffeine (1,3,7-trimethyl xanthine) were tested to confirm the specificity of the method. The derivatisation of all xanthines was performed with N-methyl-N-trimethylsilyl trifluroacetamide (MSTFA). The internal standard used was 2-(13)C ,1,3-(15)N2-theophylline. The extraction and derivatisation procedures were examined in detail and optimised stepwise during the development of the method. High-performance liquid chromatography (HPLC) was used for comparison.

  18. Determination of caffeine and associated compounds in food, beverages, natural products, pharmaceuticals, and cosmetics by micellar electrokinetic capillary chromatography.

    Science.gov (United States)

    Injac, Rade; Srdjenovic, Branislava; Prijatelj, Matevz; Boskovic, Marija; Karljikovic-Rajic, Katarina; Strukelj, Borut

    2008-02-01

    A method is described for quantitating caffeine, theobromine, theophylline, paracetamol, propyphenazone, acetylsalicylic acid, salicylic acid, and codeine phosphate in corresponding real samples of food, beverages, natural products, pharmaceuticals, and cosmetic preparations by micellar electrokinetic capillary chromatography. The separation is carried out at 25 degrees C and 25 kV, using a 20 mM phosphate buffer (pH 9.0), 80 mM sodium dodecyl sulfate, and 7.5% (v/v) acetonitrile. UV detection is at 210 nm. The method is shown to be specific, accurate (recoveries over the range 98.9-101.2%), linear over the tested range (correlation coefficients>or=0.9993), and precise (relative standard deviation below 2.1%). The method is applied for the quantitative analysis of these compounds in different foods, beverages, natural products, pharmaceuticals, and cosmetic products.

  19. Composição química de amêndoas fermentadas de cacau Chemical, composition of fermented cacao beans

    Directory of Open Access Journals (Sweden)

    Dayse Soave Spoladore

    1983-01-01

    Full Text Available During the manufacturing process 10-12% in weight of cacao beans, which consist of seed coat and embryo, are discarded. The above mentioned material has a potential for utilization as animal food and purine base source. Therefore the chemical composition was determined in each part of the beans (seed coat, embryo and cotyledons after fermentation and drying. The principal compounds of the cotyledons are lipids (48% of which 57% are saturated fatty acids. Total nitrogen, total sugar, and starch content in the seed coat and embryo varied between 3-5%, 5-6%, 11-24%, whereas crude fiber content was 20%. The theobromine and caffeine content varied between 0.6-0.9% and in the cotyledons around 0.5%.

  20. Cacao use and the San Lorenzo Olmec

    Science.gov (United States)

    Powis, Terry G.; Cyphers, Ann; Gaikwad, Nilesh W.; Grivetti, Louis; Cheong, Kong

    2011-01-01

    Mesoamerican peoples had a long history of cacao use—spanning more than 34 centuries—as confirmed by previous identification of cacao residues on archaeological pottery from Paso de la Amada on the Pacific Coast and the Olmec site of El Manatí on the Gulf Coast. Until now, comparable evidence from San Lorenzo, the premier Olmec capital, was lacking. The present study of theobromine residues confirms the continuous presence and use of cacao products at San Lorenzo between 1800 and 1000 BCE, and documents assorted vessels forms used in its preparation and consumption. One elite context reveals cacao use as part of a mortuary ritual for sacrificial victims, an event that occurred during the height of San Lorenzo's power. PMID:21555564

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

    Directory of Open Access Journals (Sweden)

    Mirian Perez Maluf

    2009-01-01

    Full Text Available 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.

  2. Inhibition of gamma-ray dose-rate effects by D/sup 2/O and inhibitors of poly(ADP-ribose) synthetase in cultured mammalian L5178Y cells

    Energy Technology Data Exchange (ETDEWEB)

    Ueno, A.M.; Tanaka, O.; Matsudaira, H.

    1984-06-01

    Effects of deuterium oxide (D/sub 2/O) and 3-aminobenzamide, an inhibitor of poly(ADP-ribose) synthetase, on cell proliferation and survival were studied in cultured mammalian L5178Y cells under growing conditions and after acute and low-dose-rate irradiation at about 0.1 to 0.4 Gy/hr of ..gamma.. rays. Growth of irradiated and unirradiated cells was inhibited by 45% D/sub 2/O but not by 3-aminobenzamide at 10mM, except for treatments longer than 30 hr. The presence of these agents either alone or in combination during irradiation at low dose rates suppressed almost totally the decrease in cell killing due to the decrease in dose rate. Among other inhibitors tested, theobromine and theophylline were found to be effective in eliminating the dose-rate effects of ..gamma.. rays. Possible mechanisms underlying the inhibition are discussed.

  3. Ritual Black Drink consumption at Cahokia

    Science.gov (United States)

    Crown, Patricia L.; Emerson, Thomas E.; Gu, Jiyan; Hurst, W. Jeffrey; Pauketat, Timothy R.; Ward, Timothy

    2012-01-01

    Chemical analyses of organic residues in fragments of pottery from the large site of Cahokia and surrounding smaller sites in Illinois reveal theobromine, caffeine, and ursolic acid, biomarkers for species of Ilex (holly) used to prepare the ritually important Black Drink. As recorded during the historic period, men consumed Black Drink in portions of the American Southeast for ritual purification. This first demonstrated discovery of biomarkers for Ilex occurs in beaker vessels dating between A.D. 1050 and 1250 from Cahokia, located far north of the known range of the holly species used to prepare Black Drink during historic times. The association of Ilex and beaker vessels indicates a sustained ritual consumption of a caffeine-laced drink made from the leaves of plants grown in the southern United States. PMID:22869743

  4. Study for luminescence performance of three methyl xanthine derivatives

    Science.gov (United States)

    Wei, Yan-Li; Dong, Chuan; Shuang, Shao-Min; Liu, Dian-Sheng

    2005-09-01

    In this paper, the low-temperature phosphorescence (LTP), the low-temperature fluorescence (LTF), the paper substrate room-temperature phosphorescence (PS-RTP) and the room fluorescence (RTF) properties of caffeine (CF), theophylline (TP), and theobromine (TB) are investigated and compared, and some rules are found out: their maximal excitation wavelength and emission wavelength are in the range of 270-300 nm and 395-445 nm, respectively. And the PS-RTP characters of lifetime, polarization and quanta yield are also investigated and compared. It is found that their lifetimes of PS-RTP are all in the level of 0.1 s. They belong to long-life phosphorescence and their PS-RTP spectra are incompletely polarized.

  5. Production and characterization of a genetically engineered anti-caffeine camelid antibody and its use in immunoaffinity chromatography.

    Science.gov (United States)

    Franco, Elliott J; Sonneson, Gregory J; DeLegge, Thomas J; Hofstetter, Heike; Horn, James R; Hofstetter, Oliver

    2010-01-15

    This work demonstrates the feasibility of using a camelid single domain antibody for immunoaffinity chromatographic separation of small molecules. An anti-caffeine VHH antibody was produced by grafting the complementarity determining sequences of a previously generated antibody onto an anti-RNase A antibody scaffold, followed by expression in E. coli. Analysis of the binding properties of the antibody by ELISA and fluorescence-based thermal shift assays showed that it recognizes not only caffeine, but also theophylline, theobromine, and paraxanthine, albeit with lower affinity. Further investigation of the effect of environmental conditions, i.e., temperature, pH, and ionic strength, on the antibody using these methods provided useful information about potential elution conditions to be used in chromatographic applications. Immobilization of the VHH onto a high flow-through synthetic support material resulted in a stationary phase capable of separating caffeine and its metabolites. 2009 Elsevier B.V. All rights reserved.

  6. Nanoparticles in plant extracts--factors which influence the formation of nanoparticles in black tea infusions.

    Science.gov (United States)

    Gröning, R; Breitkreutz, J; Baroth, V; Müller, R S

    2001-10-01

    The influence of different factors on the formation of nanoparticles in freshly brewed tea extracts was investigated. A black tea infusion was observed during cooling using photon correlation spectroscopy (PCS). The mean particle size and the number of the nanoparticles increase with decreasing temperature. In the presence of caffeine more particles are formed within the infusion. To study the influence of slight structural differences between methylxanthines, the effect of the addition of caffeine to solutions of freshly prepared decaffeinated tea was compared to that of theophylline and theobromine. In the case of theophylline fewer nanoparticles were formed. Molecular modelling calculations were performed to evaluate the most probable geometries for caffeine-polyphenol complexes. A parallel position and a congruent orientation of the 6-membered ring of caffeine and the aromatic galloyl group is the most probable geometry.

  7. Fluorescence quenching of human serum albumin by xanthines.

    Science.gov (United States)

    González-Jimènez, J; Frutos, G; Cayre, I

    1992-08-18

    A study of the fluorescence quenching of human serum albumin (HSA) by caffeine, theophylline and theobromine, based on temperature dependence, has shown that it is predominantly static. This quenching mechanism is due to the formation of a xanthine-HSA non-fluorescent complex. The Stern-Volmer equation let us determine the association constants. It seems that the quenching of the protein fluorescence depends on the number and position of the methyl groups. The temperature dependence of the association constant is used to estimate the values of the thermodynamic parameters involved in the interaction of the drugs with HSA. All three binding processes are exothermic and probably hydrophobic, and hydrogen bonds play a significant role in the stabilization of such complexes. The enthalpy and entropy changes observed appear to compensate each other to produce a relatively small Gibbs free energy.

  8. Effects of ephedrine/xanthines on thermogenesis and cold tolerance.

    Science.gov (United States)

    Vallerand, A L

    1993-02-01

    This paper reviews the use of ephedrine (E) and xanthines (X) to improve thermogenesis and cold tolerance. Recent experiments in cold-exposed subjects have shown that the beneficial effects of ingesting an E/caffeine (C) capsule on metabolic rate (M), heat debt, deep body core temperature (Tre) (P theobromine-based commercial sports bar (Cold Buster) has been similarly shown to reduce the drop in Tre. However, such a claim could not be confirmed in our lab, even in two studies performed under different environmental conditions. Despite an increase in M in some studies, C had no effect on Tre in the cold. It is concluded that E/X represent, at the moment, the best pharmacological agents to enhance cold thermogenesis and to delay the onset of hypothermia in humans.

  9. Pressurized liquid extraction of mate tea leaves.

    Science.gov (United States)

    Jacques, Rosângela Assis; Dariva, Cláudio; de Oliveira, José Vladimir; Caramão, Elina Bastos

    2008-09-05

    The objective of this work is to investigate the influence of process parameters on the pressurized liquid extraction (PLE) of Ilex paraguariensis leaves. A factorial 2(6-2) experimental design was employed using responses as the extraction yield and the chromatographic profile of the extracts. The extraction time, polarity of solvent, amount of sample, numbers of PLE cycles, flushing volume and extraction temperature were selected as independent variables (factors). Results obtained indicated that the solvent polarity was the most significant variable in the study, while the amount of sample and extraction temperature also showed significant effect. The other variables did not present significant influence in the yield of extraction. GC/MS analysis of the extract enabled the identification of saturated hydrocarbons, fatty acids, fatty acid methyl esters, phytosterols and theobromine in the extracts. Quantitative analysis of four compounds presented in the extracts (caffeine, phytol, vitamin E and squalene) was performed by the GC/MS in the SIM mode.

  10. Development and validation of a rapid ultra high pressure liquid chromatographic method for the determination of methylxanthines in herbal infusions.

    Science.gov (United States)

    Zacharis, Constantinos K; Kika, Fotini S; Tzanavaras, Paraskevas D; Fytianos, Konstantinos

    2013-05-15

    An ultra high pressure liquid chromatographic method coupled with diode array detector (UHPLC-DAD) has been developed and validated for the fast separation and determination of three major methylxanthines, i.e., caffeine, theophylline and theobromine, in various herbal beverages. Isocratic elution using 0.1vol% formic acid/CH3OH (92.5:7.5, v/v) enabled the completion of the separation cycle in less than 3min using a flow rate of 0.7mL/min and a column temperature of 50°C. Validation of the method included linearity (0.5-50mg/L), limits of detection (12-35μg/L) and quantification (40-120μg/L), precision, matrix effect and accuracy. The percent recoveries ranged between 90 and 108%. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Variation of theanine, phenolic, and methylxanthine compounds in 21 cultivars of Camellia sinensis harvested in different seasons.

    Science.gov (United States)

    Fang, Rui; Redfern, Sally P; Kirkup, Don; Porter, Elaine A; Kite, Geoffrey C; Terry, Leon A; Berry, Mark J; Simmonds, Monique S J

    2017-04-01

    This is the first study to use chemometric methods to differentiate among 21 cultivars of Camellia sinensis from China and between leaves harvested at different times of the year using 30 compounds implicated in the taste and quality of tea. Unique patterns of catechin derivatives were observed among cultivars and across harvest seasons. C. sinensis var. pubilimba (You 510) differed from the cultivars of C. sinensis var. sinensis, with higher levels of theobromine, (+)-catechin, gallocatechin, gallocatechin gallate and theasinensin B, and lower levels of (-)-epicatechin, (-)-epigallocatechin (EGC) and (-)-epigallocatechin gallate (EGCG), respectively. Three cultivars of C. sinensis var. sinensis, Fuyun 7, Qiancha 7 and Zijuan contained significantly more caffeoylquinic acids than others cultivars. A Linear Discriminant Analysis model based on the abundance of 12 compounds was able to discriminate amongst all 21 tea cultivars. Harvest time impacted the abundance of EGC, theanine and afzelechin gallate. Copyright © 2016. Published by Elsevier Ltd.

  12. Long-term functional side-effects of stimulants and sedatives in Drosophila melanogaster.

    Directory of Open Access Journals (Sweden)

    Kennedy Matsagas

    Full Text Available BACKGROUND: Small invertebrate animals, such as nematodes and fruit flies, are increasingly being used to test candidate drugs both for specific therapeutic purposes and for long-term health effects. Some of the protocols used in these experiments feature such experimental design features as lifelong virginity and very low densities. By contrast, the ability of both fruit flies and nematodes to resist stress is frequently correlated with their longevity and other functional measures, suggesting that low-stress assays are not necessarily the only useful protocol for testing the long-term effects of drugs. METHODOLOGY/PRINCIPAL FINDINGS: Here we report an alternative protocol for fruit fly drug-testing that maximizes reproductive opportunities and other types of interaction, with moderately high population densities. We validate this protocol using two types of experimental tests: 1. We show that this protocol detects previously well-established genetic differences between outbred fruit fly populations. 2. We show that this protocol is able to distinguish among the long-term effects of similar types of drugs within two broad categories, stimulants and tranquilizers. CONCLUSIONS: Large-scale fly drug testing can be conducted using mixed-sex high-density cage assays. We find that the commonly-used stimulants caffeine and theobromine differ dramatically in their chronic functional effects, theobromine being more benign. Likewise, we find that two generic pharmaceutical tranquilizers, lithium carbonate and valproic acid, differ dramatically in their chronic effects, lithium being more benign. However, these findings do not necessarily apply to human subjects, and we thus do not recommend the use of any one substance over any other.

  13. Relation of 24-hour urinary caffeine and caffeine metabolite excretions with self-reported consumption of coffee and other caffeinated beverages in the general population.

    Science.gov (United States)

    Petrovic, Dusan; Estoppey Younes, Sandrine; Pruijm, Menno; Ponte, Belén; Ackermann, Daniel; Ehret, Georg; Ansermot, Nicolas; Mohaupt, Markus; Paccaud, Fred; Vogt, Bruno; Pechère-Bertschi, Antoinette; Martin, Pierre-Yves; Burnier, Michel; Eap, Chin B; Bochud, Murielle; Guessous, Idris

    2016-01-01

    Caffeine intake is generally estimated by self-reported consumption, but it remains unclear how well self-report associates with metabolite urinary excretion. We investigated the associations of self-reported consumption of caffeinated drinks with urinary excretion of caffeine and its major metabolites in an adult population. We used data from the population-based Swiss Kidney Project on Genes in Hypertension (SKIPOGH) study. Consumption of caffeinated coffee, decaffeinated coffee and other caffeinated beverages was assessed by self-administered questionnaire. Quantification of caffeine, paraxanthine, theobromine and theophylline was performed by ultra-high performance liquid chromatography tandem mass spectrometry in 24-h urine. Association of reported consumption of caffeinated drinks with urinary caffeine derived metabolites was determined by quantile regression. We then explored the association between urinary metabolite excretion and dichotomized weekly consumption frequency of caffeinated coffee, with Receiver Operator Characteristic (ROC) analysis. In the present analysis, we included 598 individuals (52% women, mean age =46 ± 17 years). Self-reported caffeinated coffee intake was positively associated with 24-h urinary excretions of paraxanthine, theophylline and caffeine (p intakes of decaffeinated coffee and other caffeinated beverages showed no association. In ROC analysis, optimal discrimination between individuals consuming less than one caffeinated coffee/week, vs. at least one coffee, was obtained for 24-h urinary paraxanthine (Area Under Curve (AUC) = 0.868, 95% Confidence Interval (CI) [0.830;0.906]), with slightly lower performance for theophylline and caffeine, whereas theobromine did not allow any discrimination. Our results suggest that reported consumption of caffeinated coffee is positively associated with 24-h urinary excretion of caffeine, paraxanthine, and theophylline, and may be used as a marker of caffeine intake for

  14. Methylxanthine and catechin content of fresh and fermented cocoa beans, dried cocoa beans, and cocoa liquor

    Directory of Open Access Journals (Sweden)

    Pedro P. Peláez

    2016-01-01

    Full Text Available The theobromine and catechin content can affect the quality of cocoa liquor and is influenced by cacao variety, production area (PA, and fermentation, as well as the method of drying beans (FDB and cocoa liquor production (CLP. This study examined variationsin methylxanthine and catechin levels in fresh and fermented cocoa beans, dried cocoa grains, and in cocoa liquor from Trinitario, Criollo, and Forastero cacao varieties. A total of 123 cocoa bean samples from three Peruvian PAs at different altitudes, Tingo María (TM, San Alejandro (SA, and Curimana (CU, were evaluated. The theobromine (Tb and caffeine (Cf contents in fresh cocoa beans were affected by both cocoa type and PA. The caffeine content was higher in Trinitario cacao than in Criollo and Forastero varieties (p ≤ 0.05. The Tb and CF contents decreased in dry cocoa grain and was affected by FDB (p ≤ 0.05 (1.449 ± 0.004 to 1.140 ± 0.010 and 0.410 ± 0.03 to 0.165 ± 0.02 g Tb and C, respectively, per 100 g dry weight. Cocoa beans from Tingo María, which has thehighest altitude, had higher Tb and CF contents than those from other PAs. The catechin (C and epicatechin (EC contents were affected by the FDB and CLP, and were highestin fresh cocoa beans from the Tingo María area (range: 0.065 ± 0.01 to 0.020 ± 0.00 g C/100 g. The C and EC contents decreased during FDB and CLP (0.001 g C/100 g of cocoa liquor. Taken together, these results show that higher concentrations of Tb, Cf, C,and EC are present in fresh cocoa beans. Moreover, the cocoa variety influenced cocoa liquor quality. Overall, cocoa from the Tingo María PA had the most desirable chemical composition.

  15. Methylxanthines enhance the effects of cocoa flavanols on cardiovascular function: randomized, double-masked controlled studies.

    Science.gov (United States)

    Sansone, Roberto; Ottaviani, Javier I; Rodriguez-Mateos, Ana; Heinen, Yvonne; Noske, Dorina; Spencer, Jeremy P; Crozier, Alan; Merx, Marc W; Kelm, Malte; Schroeter, Hagen; Heiss, Christian

    2017-02-01

    Cocoa flavanol intake, especially that of (-)-epicatechin, has been linked to beneficial effects on human cardiovascular function. However, cocoa also contains the methylxanthines theobromine and caffeine, which may also affect vascular function. We sought to determine whether an interaction between cocoa flavanols and methylxanthines exists that influences cocoa flavanol-dependent vascular effects. Test drinks that contained various amounts of cocoa flavanols (0-820 mg) and methylxanthines (0-220 mg), either together or individually, were consumed by healthy volunteers (n = 47) in 4 different clinical studies-3 with a randomized, double-masked crossover design and 1 with 4 parallel crossover studies. Vascular status was assessed by measuring flow-mediated vasodilation (FMD), brachial pulse wave velocity (bPWV), circulating angiogenic cells (CACs), and blood pressure before and 2 h after the ingestion of test drinks. Although cocoa flavanol intake increased FMD 2 h after intake, the consumption of cocoa flavanols with methylxanthines resulted in a greater enhancement of FMD. Methylxanthine intake alone did not result in statistically significant changes in FMD. Cocoa flavanol ingestion alone decreased bPWV and diastolic blood pressure and increased CACs. Each of these changes was more pronounced when cocoa flavanols and methylxanthines were ingested together. It is important to note that the area under the curve of the plasma concentration of (-)-epicatechin metabolites over time was higher after the co-ingestion of cocoa flavanols and methylxanthines than after the intake of cocoa flavanols alone. Similar results were obtained when pure (-)-epicatechin and the methylxanthines theobromine and caffeine were consumed together. A substantial interaction between cocoa flavanols and methylxanthines exists at the level of absorption, in which the methylxanthines mediate an increased plasma concentration of (-)-epicatechin metabolites that coincides with enhanced vascular

  16. Promotional effects of new types of additives on fat crystallization.

    Science.gov (United States)

    Yoshikawa, Shinichi; Kida, Haruyasu; Sato, Kiyotaka

    2014-01-01

    We examined the promotional effects of additives on fat crystallization, such as inorganic (talc, carbon nanotube (CNT), and graphite) and organic (theobromine, ellagic acid dihydrate (EAD), and terephthalic acid) materials. The triacylglycerols (TAGs) of trilauroylglycerol (LLL), trimyristoylglycerol (MMM), and tripalmitoylglycerol (PPP) were employed as the fats. The additives (1 wt%) were added to the molten TAGs, and then the mixtures were cooled at a rate of 1°C/min followed by heating at a rate of 5°C/min. The crystallization and melting properties were observed using differential scanning calorimetry, X-ray diffraction, and polarized optical microscope (POM). Consequently, we found that the above six additives remarkably increased the initial temperatures of crystallization (Ti) on cooling without changing the melting temperatures. For example, in the case of LLL, the increases in Ti were 2.6°C (talc), 3.9°C (CNT), 8.1°C (graphite), 1.1°C (theobromine), 2.0°C (EAD), and 6.8°C (terephthalic acid). Very similar effects were observed for the crystallization of MMM and PPP with the six additives. Furthermore, the polymorphs of the first occurring crystals were changed from metastable to more stable forms by many of these additives. The POM observation revealed that the crystallization was initiated at the surfaces of additive particles. This study has shown for the first time that the heterogeneous nucleation of fat crystals can be greatly promoted by new types of additives. Such additives have great potential to promote fat crystallization by not only hydrophobic but also hydrophilic molecular interactions between the fats and additives.

  17. Differential effects of bitter compounds on the taste transduction channels TRPM5 and IP3 receptor type 3.

    Science.gov (United States)

    Gees, Maarten; Alpizar, Yeranddy A; Luyten, Tomas; Parys, Jan B; Nilius, Bernd; Bultynck, Geert; Voets, Thomas; Talavera, Karel

    2014-05-01

    Transient receptor potential cation channel subfamily M member 5 (TRPM5) is a Ca(2+)-activated nonselective cation channel involved in the transduction of sweet, bitter, and umami tastes. We previously showed that TRPM5 is a locus for the modulation of taste perception by temperature changes, and by quinine and quinidine, 2 bitter compounds that suppress gustatory responses. Here, we determined whether other bitter compounds known to modulate taste perception also affect TRPM5. We found that nicotine inhibits TRPM5 currents with an effective inhibitory concentration of ~1.3mM at -50 mV. This effect may contribute to the inhibitory effect of nicotine on gustatory responses in therapeutic and experimental settings, where nicotine is often employed at millimolar concentrations. In addition, it implies the existence of a TRPM5-independent pathway for the detection of nicotine bitterness. Nicotine seems to act from the extracellular side of the channel, reducing the maximal whole-cell conductance and inducing an acceleration of channel closure that leads to a negative shift of the activation curve. TRPM5 currents were unaffected by nicotine's metabolite cotinine, the intensive sweetener saccharin or by the bitter xanthines caffeine, theobromine, and theophylline. We also tested the effects of bitter compounds on another essential element of the sweet taste transduction pathway, the type 3 IP3 receptor (IP3R3). We found that IP3R3-mediated Ca(2+) flux is slightly enhanced by nicotine, not affected by saccharin, modestly inhibited by caffeine, theobromine, and theophylline, and strongly inhibited by quinine. Our results demonstrate that bitter compounds have differential effects on key elements of the sweet taste transduction pathway, suggesting for heterogeneous mechanisms of bitter-sweet taste interactions.

  18. The comparison of biological activity of chocolates made by different technological procedures

    Directory of Open Access Journals (Sweden)

    Lucia Godočiková

    2016-06-01

    Full Text Available 12.00 Chocolate is one of the most consumed delicacies in the world. Nowadays, raw chocolates without vanilla or allergens are getting more attention. The aim of this study was to evaluate and compare the biological activity of different types of chocolate - cold processed chocolate and chocolate made by traditional way. Total content of fat, crude fibre, polyphenols, flavonoids, phenolic acids and metylxantines - theobromine and caffeine was evaluated. The antioxidant activity was determined by a method using DPPH radical, reducing power method and phosphomolybdenum method. Both evaluated chocolates had similar content of fat and crude fibre, but sample of chocolate made by traditional way probably due to the higher content of cocoa mass had almost two times higher content of total polyphenols, flavonoids and phenolic acids as cold processed chocolate. Also the content of theobromine and caffeine was slightly higher in chocolate made by traditional way. This sample had the highest antioxidant activity - 93.68 mg TEAC.g-1 determined by phosphomolybdenum method, while in the sample of chocolate made by cold processed way was measeured value 50.82 mg TEAC.g-1. Similarly, reducing power of chocolate made by traditional way was almost two times higher, but antioxidant activity determined with DPPH method was similar in both samples (3.58 and 3.62 mg TEAC.g-1. The antioxidants and metylxantines in chocolates determine its potential to be a significant source of biologicaly active compounds with favorable effects to human health. It can be concluded in this study, that chocolate produced by conventional production technology can have more health-promoting ingredients reserved, but more extensive researches are still needed.  12.00 Normal 0 21 false false false SK X-NONE X-NONE Normal 0 21 false false false SK X-NONE X-NONE EKSTRAKSI SELULOSA DARI POD HUSK KAKAO MENGGUNAKAN SODIUM HIDROKSIDA

    Directory of Open Access Journals (Sweden)

    Gatot Siswo Hutomo

    2013-03-01

    Full Text Available EKSTRAKSI SELULOSA DARI POD HUSK KAKAO MENGGUNAKAN SODIUMHIDROKSIDA Cellulose Extraction from Cacao Pod Husk Using Sodium Hydroxide Gatot Siswo Hutomo, Djagal Wiseso Marseno, Sri Anggrahini, Supriyanto ABSTRAK Pod husk kakao banyak mengandung komponen kimia seperti pektin, lignin, hemiselulosa dan selulosa serta beberapakomponen yang lain yaitu caffein dan theobromine. Khusus selulosa dapat dilakukan modiÞ kasi sebagai turunanselulosa yang mempunyai banyak fungsi serta dapat diaplikasikan untuk pangan. Penelitian ekstraksi selulosa dari podhusk kakao telah dilakukan. Tujuan penelitian ini adalah untuk memperoleh konsentrasi sodium hidroksida sebagaibahan untuk ekstraksi selulosa dari pod husk kakao. Selulosa pada pod husk kakao terikat sangat kuat dengan lignin,sodium hidroksida akan memutus dengan baik ikatan antara selulosa dengan lignin. Bleaching pada selulosa jugadilakukan dengan menggunakan sodium hipoklorida 3% dan sodium bisulÞ t 3% untuk meningkatkan lightness. SpektraFT-IR dan X-ray juga dilakukan untuk pendeteksian pada selulosa hasil ekstraksi. Hasil ekstraksi selulosa dari pod huskkakao menggunakan sodiumhidroksida 12% menghasilkan rendemen sekitar 26,09% (db dengan kristalinitas 27,14%,kadar abu 6,56% (db, WHC 5,87 g/g dan OHC 2,74 g/g. Dapat disimpulkan bahwa sodium hidroksida 12% adalahkonsentrasi yang paling baik untuk mengekstraksi selulosa dari pod husk kakao.Kata kunci: Selulosa, pod husk, kakao, ekstraksi ABSTRACT Cacao pod husk contains some compounds like pectin, lignin, hemicelluloses and cellulose, and other compounds suchas caffeine and theobromine. Especially for cellulose should be modiÞ ed as derivates which it have multi functionsin food application. Extraction cellulose from pod husk cacao was investigated. The aim of the research was to Þ ndthe concentration of sodium hydroxide for cellulose extraction from pod husk cacao. Bleaching for cellulose werecarried out twice using sodium hypochlorite 3% (oxydator and

  19. Learning Biochemistry by Chocolate

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    M.C Guedes

    2006-07-01

    Full Text Available Both sensations and biochemical reactions taken place or promoted during ingestion of chocolate were the motivation for  investigating  the  organic  compounds  present  in  this  source.  Cocoa  and  chocolate  are  composed  by  several substances , among them, aminoacids and alkaloids.The objective of this investigation was to purpose a contextured approach  of  biochemistry  through  the  sensations  and  reactions  involving  aminoacids,  theobromine  and  hormones. Methodology: 1. Theoretical part:  constituted  by theoretical  and tutorial classes  about aminoacids, theobromine and hormones  involved  at  the  metabolism;  2.  Questionary:  ten  questions  based  upon  theoretical  classes,  personal sensations  and  general  aspects  of chocolate;  3.Lecture:  Cientific  articles  searched  in  periodics  by  own  students  as well  as  newspaper  reports;  4.  Experimental:  Laboratory  experiments  including  extraction,  characterization, spectrometric quantification  after  specific reactions  and identification by  Rf  comparison with  standards  on TLC  from cocoa  almonds  and  both  powder  cocoa  and  chocolate.  The  study  was  applied  in  30  students  from  a  chemistry college. Results: The results pointed out to a higher frequency of the students and to a increased interest  from them by   biochemistry  issues  and  cientific  lectures,  as  well  as  a  satisfactory  acquirement  of  theoretical  and  practice knowledge of aminoacids and hormones, spectrometry and chromatography. Conclusion: A contextured approach is quite positive for learning biochemistry to chemists.

  1. Direct conversion of theophylline to 3-methylxanthine by metabolically engineered E. coli.

    Science.gov (United States)

    Algharrawi, Khalid H R; Summers, Ryan M; Gopishetty, Sridhar; Subramanian, Mani

    2015-12-21

    Methylxanthines are natural and synthetic compounds found in many foods, drinks, pharmaceuticals, and cosmetics. Aside from caffeine, production of many methylxanthines is currently performed by chemical synthesis. This process utilizes many chemicals, multiple reactions, and different reaction conditions, making it complicated, environmentally dissatisfactory, and expensive, especially for monomethylxanthines and paraxanthine. A microbial platform could provide an economical, environmentally friendly approach to produce these chemicals in large quantities. The recently discovered genes in our laboratory from Pseudomonas putida, ndmA, ndmB, and ndmD, provide an excellent starting point for precisely engineering Escherichia coli with various gene combinations to produce specific high-value paraxanthine and 1-, 3-, and 7-methylxanthines from any of the economical feedstocks including caffeine, theobromine or theophylline. Here, we show the first example of direct conversion of theophylline to 3-methylxanthine by a metabolically engineered strain of E. coli. Here we report the construction of E. coli strains with ndmA and ndmD, capable of producing 3-methylxanthine from exogenously fed theophylline. The strains were engineered with various dosages of the ndmA and ndmD genes, screened, and the best strain was selected for large-scale conversion of theophylline to 3-methylxanthine. Strain pDdA grown in super broth was the most efficient strain; 15 mg/mL cells produced 135 mg/L (0.81 mM) 3-methylxanthine from 1 mM theophylline. An additional 21.6 mg/L (0.13 mM) 1-methylxanthine were also produced, attributed to slight activity of NdmA at the N 3 -position of theophylline. The 1- and 3-methylxanthine products were separated by preparative chromatography with less than 5% loss during purification and were identical to commercially available standards. Purity of the isolated 3-methylxanthine was comparable to a commercially available standard, with no contaminant peaks as

  2. Synthesis, Characterization, and Thermal and Antimicrobial Activities of Some Novel Organotin(IV: Purine Base Complexes

    Directory of Open Access Journals (Sweden)

    Reena Jain

    2013-01-01

    Full Text Available A new series of organotin(IV complexes with purine bases theophylline (HL1 and theobromine (L2 of the types R3Sn(L1, R2Sn(L1Cl, R3Sn(L2Cl, and R2Sn(L2Cl2 (R = C6H5CH2–; p-ClC6H4CH2– have been synthesized in anhydrous THF. The complexes were characterized by elemental analysis, conductance measurements, molecular weight determinations, UV-vis, IR, 1H, 13C NMR, and mass spectral studies. Various kinetic and thermodynamic parameters of these complexes have also been determined using TG/DTA technique. The thermal decomposition techniques indicate the formation of SnO2 as a residue. The results show that the ligands act as bidentate, forming a five-member chelate ring. All the complexes are 1 : 1 metal-ligand complexes. In order to assess their antimicrobial activity, the ligands and their corresponding complexes have also been tested in vitro against bacteria (E. coli, S. aureus, and P. pyocyanea and fungi (Rhizopus oryzae and Aspergillus flavus. All the complexes exhibit remarkable activity, and the results provide evidence that the studied complexes might indeed be a potential source of antimicrobial agents.

  3. Supraventricular tachycardia induced by chocolate: is chocolate too sweet for the heart?

    Science.gov (United States)

    Parasramka, Saurabh; Dufresne, Alix

    2012-09-01

    Conflicting studies have been published concerning the association between chocolate and cardiovascular diseases. Fewer articles have described the potential arrhythmogenic risk related to chocolate intake. We present a case of paroxysmal supraventricular tachycardia in a woman after consumption of large quantity of chocolate. A 53-year-old woman with no significant medical history presented to us with complaints of palpitations and shortness of breath after consuming large amounts of chocolate. Electrocardiogram showed supraventricular tachycardia at 165 beats per minute, which was restored to sinus rhythm after adenosine bolus injection. Electrophysiology studies showed atrioventricular nodal reentry tachycardia, which was treated with radiofrequency ablation. Chocolate contains caffeine and theobromine, which are methylxanthines and are competitive antagonists of adenosine and can have arrhythmogenic potential. Our case very well describes an episode of tachycardia precipitated by large amount of chocolate consumption in a patient with underlying substrate. There are occasional case reports describing association between chocolate, caffeine, and arrhythmias. A large Danish study, however, did not find any association between amount of daily caffeine consumption and risk of arrhythmia.

  4. Validated high-performance liquid chromatographic method for the standardisation of Ptychopetalum olacoides Benth., Olacaceae, commercial extracts

    Directory of Open Access Journals (Sweden)

    Renata Colombo

    2010-10-01

    Full Text Available Ptychopetalum olacoides Benth., Olacaceae, popularly known as marapuama or muirapuama or miriantã, is a species native to the Amazonian region of Brazil. Extracts of the bark of the plant have been used traditionally for its stimulating and aphrodisiac properties and currently commercialised by the herbal industry as constituents in a wide range of phytomedicines. Fractionation by open column chromatography followed by preparative HPLC-UV/PAD of the stem bark and of three commercial extracts of P. olacoides allowed the isolation of three components that were common to all extracts analysed, and these were identified by NMR to be vanillic acid, protocatechuic acid and theobromine. Vanillic acid, which has been proposed as a phytochemical marker for P. olacoides, was employed as an external standard in the development and validation of a rapid qualitative and quantitative HPLC assay for the analyte. The recoveries values of the developed method were 99.02% and the LOD and LOQ values were 0.033 and 0.11 mg.L-1, respectively. The described method may be applied to the standardisation of herbs, extracts or phytomedicines commercialised as marapuama.

  5. Alkaloids and athlete immune function: caffeine, theophylline, gingerol, ephedrine, and their congeners.

    Science.gov (United States)

    Senchina, David S; Hallam, Justus E; Kohut, Marian L; Nguyen, Norah A; Perera, M Ann d N

    2014-01-01

    Plant alkaloids are found in foods, beverages, and supplements consumed by athletes for daily nutrition, performance enhancement, and immune function improvement. This paper examined possible immunomodulatory roles of alkaloids in exercise contexts, with a focus on human studies. Four representative groups were scrutinized: (a) caffeine (guaranine, mateine); (b) theophylline and its isomers, theobromine and paraxanthine; (c) ginger alkaloids including gingerols and shogaol; and (d) ephedra alkaloids such as ephedrine and pseudoephedrine. Emerging or prospective alkaloid sources (Goji berry, Noni berry, and bloodroot) were also considered. Human in vitro and in vivo studies on alkaloids and immune function were often conflicting. Caffeine may be immunomodulatory in vivo depending on subject characteristics, exercise characteristics, and immune parameters measured. Caffeine may exhibit antioxidant capacities. Ginger may exert in vivo anti-inflammatory effects in certain populations, but it is unclear whether these effects are due to alkaloids or other biochemicals. Evidence for an immunomodulatory role of alkaloids in energy drinks, cocoa, or ephedra products in vivo is weak to nonexistent. For alkaloid sources derived from plants, variability in the reviewed studies may be due to the presence of unrecognized alkaloids or non-alkaloid compounds (which may themselves be immunomodulatory), and pre-experimental factors such as agricultural or manufacturing differences. Athletes should not look to alkaloids or alkaloid-rich sources as a means of improving immune function given their inconsistent activities, safety concerns, and lack of commercial regulation.

  6. Does the chocolate interfere with the radiolabelling of erythrocytes with 99 mTc?; Le chocolat interfere-t-il avec le radiomarquage des erythrocytes au 99 mTc?

    Energy Technology Data Exchange (ETDEWEB)

    Bustani, H.; Colavolpe, C.; Imbert-Joscht, I.; Moubarik, C.; Havlik, P.; Pisano, P.; Guillet, B. [Service de medecine nucleaire, CHU Nord, Marseille, (France)

    2009-05-15

    We present in this work the case of a failure of erythrocytes labelling with {sup 99m}Tc for a twenty five years old patient making the examination not interpretable. The patient reported that she drank a chocolate drink the morning before the examination. it is the first observation of a such interaction between the chocolate consumption. We do not have any explanation at this date, some compounds in the cocoa can be considered with caution. Some flavonoids (catechins and pro-cyanidins) modify the plasmatic and intra-erythrocyte oxido-reducer statute and could interact with the labelling (specific reduction of the {sup 99m}Tc pertechnetate in the middle of erythrocyte by the tin pyrophosphate. These compounds, as well as the methyl-xanthine-theobromine, seem modify the membrane permeability of erythrocytes and could interfere with the input of pyrophosphate or {sup 99m}Tc-pertechnetate in the middle of erythrocyte. these observations, in spite of preliminary ones, lead us to recommend near the patients to avoid the chocolate foods in the twenty four hours before this type of examinations. (N.C.)

  7. Development of urine standard reference materials for metabolites of organic chemicals including polycyclic aromatic hydrocarbons, phthalates, phenols, parabens, and volatile organic compounds.

    Science.gov (United States)

    Schantz, Michele M; Benner, Bruce A; Heckert, N Alan; Sander, Lane C; Sharpless, Katherine E; Vander Pol, Stacy S; Vasquez, Y; Villegas, M; Wise, Stephen A; Alwis, K Udeni; Blount, Benjamin C; Calafat, Antonia M; Li, Zheng; Silva, Manori J; Ye, Xiaoyun; Gaudreau, Éric; Patterson, Donald G; Sjödin, Andreas

    2015-04-01

    Two new Standard Reference Materials (SRMs), SRM 3672 Organic Contaminants in Smokers' Urine (Frozen) and SRM 3673 Organic Contaminants in Non-Smokers' Urine (Frozen), have been developed in support of studies for assessment of human exposure to select organic environmental contaminants. Collaborations among three organizations resulted in certified values for 11 hydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) and reference values for 11 phthalate metabolites, 8 environmental phenols and parabens, and 24 volatile organic compound (VOC) metabolites. Reference values are also available for creatinine and the free forms of caffeine, theobromine, ibuprofen, nicotine, cotinine, and 3-hydroxycotinine. These are the first urine Certified Reference Materials characterized for metabolites of organic environmental contaminants. Noteworthy, the mass fractions of the environmental organic contaminants in the two SRMs are within the ranges reported in population survey studies such as the National Health and Nutrition Examination Survey (NHANES) and the Canadian Health Measures Survey (CHMS). These SRMs will be useful as quality control samples for ensuring compatibility of results among population survey studies and will fill a void to assess the accuracy of analytical methods used in studies monitoring human exposure to these organic environmental contaminants.

  8. EFFECT OF FERMENTED CACAO POD SUPPLEMENTATION ON SHEEP RUMEN MICROBIAL FERMENTATION

    Directory of Open Access Journals (Sweden)

    S. Wulandari

    2015-09-01

    Full Text Available The objective of this research was to improve beneficial value of cacao pod as sheep feedingredients comprising up to 50% total feed. This research was conducted in two stages. Stage 1 wascacao pod fermentation. Completely randomized design with 3x3 factorial patterns was used in thisstage, in which factor I was microbial inoculum dosage of 0%, 0.05% and 0.1% and factor II wasincubation period of 0, 3 and 6 days. Result demonstrated that six-day fermentation with 0.05%microbial inoculum could lower cacao NDF, ADF and theobromine. The optimum inoculum dosage andfermentation time from stage 1 was applied to stage 2. Stage 2 was rumen microbial fermentation test.This research administrated 3x3 of latin square design. In period I sheep were fed with CF0 (nonfermentedcomplete feed, in period II sheep were given CF 1 (complete feed containing fermentedcacao pod and in period III sheep were given CF2 (fermented complete feed based cacao pod. Resultdemonstrated that pH value of sheep microbial liquid in treatment of CF0, CF1 and CF2 was in normalpH range and did not affect volatile fatty acids (VFA and ammonia. In conclusion, supplementing up to 50% of feed with complete feed containing fermented or non-fermented cacao pod did not affect theprocess of rumen microbial fermentation.

  9. The effect of flavanol-rich cocoa on cerebral perfusion in healthy older adults during conscious resting state: a placebo controlled, crossover, acute trial.

    Science.gov (United States)

    Lamport, Daniel J; Pal, Deepa; Moutsiana, Christina; Field, David T; Williams, Claire M; Spencer, Jeremy P E; Butler, Laurie T

    2015-09-01

    There has recently been increasing interest in the potential of flavanols, plant-derived compounds found in foods such as fruit and vegetables, to ameliorate age-related cognitive decline. Research suggests that cocoa flavanols improve memory and learning, possibly as a result of their anti-inflammatory and neuroprotective effects. These effects may be mediated by increased cerebral blood flow (CBF), thus, stimulating neuronal function. The present study employed arterial spin labelling functional magnetic resonance imaging to explore the effect of a single acute dose of cocoa flavanols on regional CBF. CBF was measured pre- and post-consumption of low (23 mg) or high (494 mg) 330 ml equicaloric flavanol drinks matched for caffeine, theobromine, taste and appearance according to a randomized counterbalanced crossover double-blind design in eight males and ten females, aged 50-65 years. Changes in perfusion from pre- to post-consumption were calculated as a function of each drink. Significant increases in regional perfusion across the brain were observed following consumption of the high flavanol drink relative to the low flavanol drink, particularly in the anterior cingulate cortex and the central opercular cortex of the parietal lobe. Consumption of cocoa flavanol improves regional cerebral perfusion in older adults. This provides evidence for a possible acute mechanism by which cocoa flavanols are associated with benefits for cognitive performance.

  10. Identification and determination of synthetic pharmaceuticals as adulterants in eight common herbal weight loss supplements.

    Science.gov (United States)

    Khazan, Marjan; Hedayati, Mehdi; Kobarfard, Farzad; Askari, Sahar; Azizi, Fereidoun

    2014-03-01

    Adulterated herbal weight loss products with containing undeclared synthetic drugs are common and responsible for many serious health damages. The purpose of the study was to determine five synthetic adulterants in eight common herbal weight loss supplements, which are currently sold in Iran markets, to verify their presence in supplements, without mentioning on the labels. Eight common herbal weight loss samples were obtained from the Iran pharmaceutical market after advertising in the Persian language on satellite channels and internet. Five pharmacological classes of drugs used for weight loss, namely sibutramine, phenolphthalein, phenytoin, bumetanide and rimonabant, were investigated and quantified by GC-MS for the first three and LC-MS for the last two medications. The most undeclared ingredients, which were illegally added include sibutramine, phenolphthalein, bumetanide, and phenytoin in the original super slim, herbaceous essence, super slim green lean, and fat loss, supplements, respectively. Rimonabant was not found. Caffeine, pseudoephedrine, theobromine and amfepramone were also found in the supplements using GC-MS assay. Adulterated synthetic substances were detected in the herbal weight loss products. Health care professionals should make people aware of the risks of taking herbal weight-loss supplements.

  11. Application of Laser Mass Spectrometry to Art and Archaeology

    Science.gov (United States)

    Gulian, Lase Lisa E.; Callahan, Michael P.; Muliadi, Sarah; Owens, Shawn; McGovern, Patrick E.; Schmidt, Catherine M.; Trentelman, Karen A.; deVries, Mattanjah S.

    2011-01-01

    REMPI laser mass spectrometry is a combination of resonance enhanced multiphoton ionization spectroscopy and time of flight mass spectrometry, This technique enables the collection of mass specific optical spectra as well as of optically selected mass spectra. Analytes are jet-cooled by entrainment in a molecular beam, and this low temperature gas phase analysis has the benefit of excellent vibronic resolution. Utilizing this method, mass spectrometric analysis of historically relevant samples can be simplified and improved; Optical selection of targets eliminates the need for chromatography while knowledge of a target's gas phase spectroscopy allows for facile differentiation of molecules that are in the aqueous phase considered spectroscopically indistinguishable. These two factors allow smaller sample sizes than commercial MS instruments, which in turn will require less damage to objects of antiquity. We have explored methods to optimize REMPI laser mass spectrometry as an analytical tool to archaeology using theobromine and caffeine as molecular markers in Mesoamerican pottery, and are expanding this approach to the field of art to examine laccaic acid in shellacs.

  12. The effects of maternal caffeine and chocolate intake on fetal heart rate.

    Science.gov (United States)

    Buscicchio, Giorgia; Piemontese, Mariangela; Gentilucci, Lucia; Ferretti, Filippo; Tranquilli, Andrea L

    2012-05-01

    The aim of our study was to analyze the effects of caffeine and chocolate (70% cocoa) on fetal heart rate (FHR). Fifty pregnant women with uncomplicated gestation, matched for age and parity, underwent computerized FHR recording before and after the consumption of caffeine and then, after one week, before and after 70% cocoa chocolate intake. Computerized cardiotocography (cCTG) parameters were expressed as mean and SD. The differences were tested for statistical significance using the paired t-test, with significance at p intake. The number of large accelerations, the duration of episodes of high variation and the short-term FHR variation were significantly higher (p consumption of chocolate, whilst no effect of cocoa was found during contractions. Our results suggest that maternal intake of both caffeine and 70% cocoa have a stimulating action on fetal reactivity. This finding is likely due to the pharmacological action of theobromine, a methilxanthine present in coffee and in chocolate. The correlation between maternal caffeine intake and increased uterine contraction peaks is likely due to the effect of caffeine on the uterine muscle.

  13. Genome-wide association study of caffeine metabolites provides new insights to caffeine metabolism and dietary caffeine-consumption behavior.

    Science.gov (United States)

    Cornelis, Marilyn C; Kacprowski, Tim; Menni, Cristina; Gustafsson, Stefan; Pivin, Edward; Adamski, Jerzy; Artati, Anna; Eap, Chin B; Ehret, Georg; Friedrich, Nele; Ganna, Andrea; Guessous, Idris; Homuth, Georg; Lind, Lars; Magnusson, Patrik K; Mangino, Massimo; Pedersen, Nancy L; Pietzner, Maik; Suhre, Karsten; Völzke, Henry; Bochud, Murielle; Spector, Tim D; Grabe, Hans J; Ingelsson, Erik

    2016-12-15

    Caffeine is the most widely consumed psychoactive substance in the world and presents with wide interindividual variation in metabolism. This variation may modify potential adverse or beneficial effects of caffeine on health. We conducted a genome-wide association study (GWAS) of plasma caffeine, paraxanthine, theophylline, theobromine and paraxanthine/caffeine ratio among up to 9,876 individuals of European ancestry from six population-based studies. A single SNP at 6p23 (near CD83) and several SNPs at 7p21 (near AHR), 15q24 (near CYP1A2) and 19q13.2 (near CYP2A6) met GW-significance (P caffeine and lower plasma paraxanthine/caffeine (slow caffeine metabolism) were previously associated with lower coffee and caffeine consumption behavior in GWAS. Variants at 19q13.2 associated with higher plasma paraxanthine/caffeine (slow paraxanthine metabolism) were also associated with lower coffee consumption in the UK Biobank (n = 94 343, P consumption in GWAS were nominally associated with plasma caffeine or its metabolites. Taken together, we have identified genetic factors contributing to variation in caffeine metabolism and confirm an important modulating role of systemic caffeine levels in dietary caffeine consumption behavior. Moreover, candidate genes identified encode proteins with important clinical functions that extend beyond caffeine metabolism. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  14. Erythema multiforme-like eruption from a slimming drug preparation cutaneous adverse drug reaction

    Directory of Open Access Journals (Sweden)

    Linda Tognetti

    2011-01-01

    Full Text Available We report a case of a 34-year-old woman presenting with an erythema multiforme (EM-like eruption. Lesions developed after a 12-day treatment with a slimming drug preparation (food integrator with thermogenic activity and a herbal remedy (pilosella tincture. Serological investigations excluded viral or bacterial infections. Patch testing with galenic preparations of both drugs demonstrated sensitization to the slimming drug preparation. According to literature reports and immune-chemical properties, those components that are likely to have triggered the skin eruption are clorazepate dipotassium and theobromine. Their interaction with other two constituents such as pseudoephedrine hydrochloride and dehydrocholic acid may have caused the adverse reaction by means of a summation effect. There are no reports specifically about EM caused by a slimming drug preparation and no studies have identified thermogenic pills as cause of EM/EM-like eruption. Weight-loss compounds in slimming preparations should be kept in mind as a possible cause of drug-induced EM-like eruption.

  15. The effects of different concentrations of cocoa in the chocolate intaken by the mother on fetal heart rate.

    Science.gov (United States)

    Buscicchio, Giorgia; Lorenzi, Sara; Tranquilli, Andrea Luigi

    2013-10-01

    To analyze the effects of different concentrations (30% and 80%) of cocoa on fetal heart rate (FHR). One hundred pregnant women with uncomplicated gestation, matched for age and parity, underwent computerized FHR recording before and after the consumption of 30 g of 30% and 80% cocoa chocolate. After 1 week, those who had received 30% were shifted to 80% and vice versa to have a crossover. Computerized cardiotocography parameters (contractions, fetal movements, baseline FHR, accelerations greater than 15 bpm for 15 s, number of decelerations, minutes of high variability, short term variability in ms) were recorded and expressed as mean and SD. The differences were tested for statistical significance using the paired t test, with the significance at p cocoa intake. After 30% cocoa chocolate intake, only the number of accelerations was significantly increased. The percent change of the number of accelerations and the short-term FHR variation were significantly higher after 80% cocoa chocolate maternal intake. Maternal intake of dark chocolate has a stimulating action on fetal reactivity. The effect is more marked with high concentrations (80%) of cocoa. This finding is likely due to the pharmacological action of theobromine, a methilxanthine present in cocoa.

  16. Multi-element, multi-compound isotope profiling as a means to distinguish the geographical and varietal origin of fermented cocoa (Theobroma cacao L.) beans.

    Science.gov (United States)

    Diomande, Didier; Antheaume, Ingrid; Leroux, Maël; Lalande, Julie; Balayssac, Stéphane; Remaud, Gérald S; Tea, Illa

    2015-12-01

    Multi-element stable isotope ratios have been assessed as a means to distinguish between fermented cocoa beans from different geographical and varietal origins. Isotope ratios and percentage composition for C and N were measured in different tissues (cotyledons, shells) and extracts (pure theobromine, defatted cocoa solids, protein, lipids) obtained from fermented cocoa bean samples. Sixty-one samples from 24 different geographical origins covering all four continental areas producing cocoa were analyzed. Treatment of the data with unsupervised (Principal Component Analysis) and supervised (Partial Least Squares Discriminant Analysis) multiparametric statistical methods allowed the cocoa beans from different origins to be distinguished. The most discriminant variables identified as responsible for geographical and varietal differences were the δ(15)N and δ(13)C values of cocoa beans and some extracts and tissues. It can be shown that the isotope ratios are correlated with the altitude and precipitation conditions found in the different cocoa-growing regions. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Involvement of a novel intronic microRNA in cross regulation of N-methyltransferase genes involved in caffeine biosynthesis in Coffea canephora.

    Science.gov (United States)

    Mohanan, Shibin; Gowda, Kalpashree; Kandukuri, Satyanarayana V; Chandrashekar, Arun

    2013-04-25

    There are numerous reports on intronic miRNAs from plants, most of which are involved in the regulation of unrelated genes. Some of the target genes are antagonistic to the host genes. Intronic miRNAs in animal systems, however, are known to have synergistic effects. This article is the first to report a similar regulatory effect of a miRNA originating from an intron in plants. NMT genes involved in caffeine biosynthesis were silenced to obtain transformants with reduced caffeine. Transcript analysis revealed the accumulation of transcripts for a related NMT gene (CaMTL1) in transformants bearing either antisense or RNAi constructs. The altered expression was assumed to relate to the silencing of the NMT genes. Bioinformatics analysis of the genes involved in biosynthesis revealed the presence of an intronic miRNA originating from the intron of the theobromine synthase gene targeting CaMTL1. The putative miRNA was cloned and sequenced. Modified 5'-RLM-RACE mapping of the cleavage site and subsequent Northern blotting experimentally demonstrated the presence and activity of such a miRNA in Coffea canephora. This novel regulatory mechanism previously unreported in plants will shed more light onto the evolution of multigene families and the role of introns in this process. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. General and Genetic Toxicology of Guayusa Concentrate (Ilex guayusa).

    Science.gov (United States)

    Kapp, Robert W; Mendes, Odete; Roy, Shambhu; McQuate, Robert S; Kraska, Richard

    2016-01-01

    Tea from the leaves of guayusa (Ilex guayusa) has a long history of consumption by Ecuadorian natives in regions where the plant is indigenous. The tea contains the methylxanthines caffeine and theobromine as well as chlorogenic acids, flavonoids, and sugars. Various studies were performed to evaluate the general and genetic toxicology of a standardized liquid concentrate of guayusa (GC). Guayusa concentrate was found to be negative in in vitro genotoxicity tests including the Ames test and a chromosome aberration study in human lymphocytes. The oral median lethal dose (LD50) of GC was >5,000 mg/kg for female rats. Guayusa concentrate was administered to male and female rats in a 90-day subchronic study at 1,200, 2,500, and 5,000 mg/kg/d of GC and a caffeine-positive control at 150 mg/kg/d corresponding to the amount of caffeine in the high-dose GC group. Effects observed in the GC-treated groups were comparable to those in the caffeine control group and included reductions in body weights, food efficiency, triglycerides values, and fat pad weights and increases in blood chemistry values for serum aspartate aminotransferase, serum alanine aminotransferase, and cholesterol and adaptive salivary gland hypertrophy. No signs of incremental toxicity due to any other components of guayusa were observed. The studies indicate no harmful effects of GC in these test systems. © The Author(s) 2016.

  19. Energy drinks: a review of use and safety for athletes.

    Science.gov (United States)

    Duchan, Erin; Patel, Neil D; Feucht, Cynthia

    2010-06-01

    Energy drinks have increased in popularity in adolescents and young adults; however, concerns have been raised regarding the ingredients in energy drinks and their potential negative effects on health. Caffeine, the most physiologically active ingredient in energy drinks, is generally considered safe by the US Food and Drug Administration (FDA), although adverse effects can occur at varying amounts. Guarana, which contains caffeine in addition to small amounts of theobromine, theophylline, and tannins, is also recognized as safe by the FDA, although it may lead to caffeine toxicity when combined with caffeine. The amount of ginseng in energy drinks is typically far below the amount used as a dietary supplement, and is generally considered safe. Taurine, an intracellular amino acid, has been reported to have positive inotropic effects; however, this claim is not supported by research. Most energy drinks also contain sugar in an amount that exceeds the maximum recommended daily amount. Young athletes are increasingly using energy drinks because of the ergogenic effects of caffeine and the other ingredients found in these beverages. Energy drinks combined with alcohol are also gaining popularity in young adults, which poses significant concerns about health risks. Other health concerns related to consumption of energy drinks include case reports of seizures and cardiac arrest following energy drink consumption and dental enamel erosion resulting from the acidity of energy drinks.

  20. Is Failure of Cocrystallization Actually a Failure? Eutectic Formation in Cocrystal Screening of Hesperetin.

    Science.gov (United States)

    Chadha, Kunal; Karan, Maninder; Chadha, Renu; Bhalla, Yashika; Vasisht, Karan

    2017-08-01

    Cocrystal screening of hesperetin with certain countermolecules generated highly soluble noncovalent derivatives in the form of eutectics, instead of expected cocrystals. As adhesive forces established by complimentary functional groups on hesperetin and coformers were unable to overcome the stress due to size shape mismatch of component molecules, thus, eutectics were formed. Hesperetin, a polyphenolic antioxidant with potent anticancer and cardioprotective effects, has an underdeveloped role in modern therapeutics on account of its critically low aqueous solubility resulting in stunted bioavailability. The liquid-assisted cogrinding of hesperetin and coformers generated binary-phase eutectics in fixed stoichiometry with theophylline (1:1.5), adenine (2:1), gallic acid (1.5:1), and theobromine (2:1). Primarily characterized by lower melting endotherm in differential scanning calorimetry, the eutectics showed complete melting in hot-stage microscopy. Apart from characteristic V-shaped binary-phase diagram, no discernible changes in the FTIR and powder X-ray diffraction spectra further confirm eutectic formation. The morphological differences were analyzed by SEM measurements. A 2 to 4 times enhanced dissolution profile of the eutectics measured in pH 7.4 aqueous buffer was coupled with the in vitro (1,1-diphenyl-2-picryl hydroxyl free radical antioxidant assay and RBC antihemolytic assay) studies to present a complete preliminary data on the improved bioavailability of hesperetin eutectics. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  1. Erythema multiforme-like eruption from a slimming drug preparation cutaneous adverse drug reaction.

    Science.gov (United States)

    Tognetti, Linda; Giorgini, Simonetta; Lotti, Torello

    2011-07-01

    We report a case of a 34-year-old woman presenting with an erythema multiforme (EM)-like eruption. Lesions developed after a 12-day treatment with a slimming drug preparation (food integrator with thermogenic activity) and a herbal remedy (pilosella tincture). Serological investigations excluded viral or bacterial infections. Patch testing with galenic preparations of both drugs demonstrated sensitization to the slimming drug preparation. According to literature reports and immune-chemical properties, those components that are likely to have triggered the skin eruption are clorazepate dipotassium and theobromine. Their interaction with other two constituents such as pseudoephedrine hydrochloride and dehydrocholic acid may have caused the adverse reaction by means of a summation effect. There are no reports specifically about EM caused by a slimming drug preparation and no studies have identified thermogenic pills as cause of EM/EM-like eruption. Weight-loss compounds in slimming preparations should be kept in mind as a possible cause of drug-induced EM-like eruption.

  2. Agrobacterium-mediated silencing of caffeine synthesis through root transformation in Camellia sinensis L.

    Science.gov (United States)

    Mohanpuria, Prashant; Kumar, Vinay; Ahuja, Paramvir Singh; Yadav, Sudesh Kumar

    2011-07-01

    Tea [Camellia sinensis (L.) O. Kuntze] is a perennial and most popular non-alcoholic caffeine-containing beverage crop. Tea has several constraints for its genetic improvement such as its high polyphenolic content and woody perennial nature. The development of transgenic tea is very difficult, laborious, and time taking process. In tea, regeneration requires minimum 8-12 months. In view of this, attempt has been made in this article to develop a rapid, efficient, and quite economical Agrobacterium-mediated root transformation system for tea. The feasibility of the developed protocol has been documented through silencing caffeine biosynthesis. For this, one-month-old tea seedlings were exposed to fresh wounding at the elongation zone of roots and were inoculated with Agrobacterium tumefaciens cultures carrying a RNAi construct (pFGC1008-CS). The pFGC1008-CS contained 376 bp of caffeine synthase (CS) cDNA fragment in sense and antisense direction with an intron in between. This has made the RNAi construct to produce a hairpin RNA (ihpRNA). The suppressed expression of CS gene and a marked reduction in caffeine and theobromine contents in young shoots of tea seedlings were obtained after root transformation through Agrobacterium infiltration. Such transformation system could be useful for functional analysis of genes in tea like woody and perennial plants.

  3. Theacrine (1,3,7,9-tetramethyluric acid) synthesis in leaves of a Chinese tea, kucha (Camellia assamica var. kucha).

    Science.gov (United States)

    Zheng, Xin-Qiang; Ye, Chuang-Xing; Kato, Misako; Crozier, Alan; Ashihara, Hiroshi

    2002-05-01

    Theacrine (1,3,7,9-tetramethyluric acid) and caffeine were the major purine alkaloids in the leaves of an unusual Chinese tea known as kucha (Camellia assamica var. kucha). Endogenous levels of theacrine and caffeine in expanding buds and young leaves were ca. 2.8 and 0.6-2.7% of the dry wt, respectively, but the concentrations were lower in the mature leaves. Radioactivity from S-adenosyl-L-[methyl-14C]methionine was incorporated into theacrine as well as theobromine and caffeine by leaf disks of kucha, indicating that S-adenosyl-L-methionine acts as the methyl donor not only for caffeine biosynthesis but also for theacrine production. [8-14C]Caffeine was converted to theacrine by kucha leaves with highest incorporation occurring in expanding buds. When [8-14C]adenosine, the most effective purine precursor for caffeine biosynthesis in tea (Camellia sinensis), was incubated with young kucha leaves for 24 h, up to 1% of total radioactivity was recovered in theacrine. However, pulse-chase experiments with [8-14C]adenosine demonstrated much more extensive incorporation of label into caffeine than theacrine, possibly because of dilution of [14C]caffeine produced by the large endogenous caffeine pool. These results indicate that in kucha leaves theacrine is synthesized from caffeine in what is probably a three-step pathway with 1,3,7-methyluric acid acting an intermediate. This is a first demonstration that theacrine is synthesized from adenosine via caffeine.

  4. Differing chemical compositions of three teas may explain their different effects on acute blood pressure in spontaneously hypertensive rats.

    Science.gov (United States)

    Li, Shan-Bing; Li, Yi-Fang; Mao, Zhong-Fu; Hu, Hui-Hua; Ouyang, Shu-Hua; Wu, Yan-Ping; Tsoi, Bun; Gong, Pin; Kurihara, Hiroshi; He, Rong-Rong

    2015-04-01

    Heavy tea consumption is suggested to be unsuitable for hypertensive people. However, the bioactive substances in different varieties of tea leaves are very different. This study compares the effects of three Chinese teas - C. sinensis, C. ptilophylla and C. assamica var. kucha - on blood pressure (BP) and heart rate in spontaneously hypertensive rats (SHRs). Intragastric administration of C. sinensis extract led to an acute increase in systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate in SHRs. However, C. ptilophylla and C. assamica var. kucha exerted no obvious influences on SBP, DBP or heart rate. Similar to the extract of C. sinensis, intragastric administration of caffeine also led to an acute increase in BP and heart rate in SHRs. In contrast, theobromine and theacrine - purine alkaloids predominantly contained in C. ptilophylla and C. assamica var. kucha, respectively - had no pressor effects. The effect of caffeine on BP was related to the regulation of plasma epinephrine and norepinephrine levels in SHRs. The different effects of C. sinensis, C. ptilophylla and C. assamica var. kucha on BP might be explained, at least partially, by the differences in the varieties and contents of purine alkaloids. © 2014 Society of Chemical Industry.

  5. Does Caffeine Consumption Modify Cerebrospinal Fluid Amyloid-β Levels in Patients with Alzheimer's Disease?

    DEFF Research Database (Denmark)

    Travassos, Maria; Santana, Isabel; Baldeiras, Inês

    2015-01-01

    Caffeine may be protective against Alzheimer's disease (AD) by modulating amyloid-β (Aβ) metabolic pathways. The present work aimed to study a possible association of caffeine consumption with the cerebrospinal fluid (CSF) biomarkers, particularly Aβ. The study included 88 patients with AD or mild...... cognitive impairment. The consumption of caffeine and theobromine was evaluated using a validated food questionnaire. Quantification of caffeine and main active metabolites was performed with liquid chromatography coupled to tandem mass spectrometry. The levels of A(1-42), total tau, and phosphorylated tau...... in the CSF were determined using sandwich ELISA methods and other Aβ species, Aβ(X-38), Aβ(X-40), and Aβ(X-42), with the MSD Aβ Triplex assay. The concentration of caffeine was 0.79±1.15 μg/mL in the CSF and 1.20±1.88 μg/mL in the plasma. No correlation was found between caffeine consumption and Aβ42...

  6. Measurement of caffeine and its three primary metabolites in human plasma by HPLC-ESI-MS/MS and clinical application.

    Science.gov (United States)

    Chen, Feng; Hu, Zhe-Yi; Parker, Robert B; Laizure, S Casey

    2017-06-01

    Caffeine is a mild stimulant with significant potential for abuse, being consumed in larger doses with the widespread availability of energy drinks and by novel routes of administration such as inspired powder, oral sprays and electronic cigarettes. How these recent changes in caffeine consumption affecting caffeine disposition and abuse potential is of growing concern. In the study of caffeine disposition in humans, it is common to only measure the caffeine concentration; however, caffeine's three major metabolites (paraxanthine, theobromine and theophylline) retain central nervous system stimulant activity that may contribute to the overall pharmacological activity and toxicity. Therefore, it would be scientifically more rigorous to measure caffeine and its major metabolites in the evaluation of caffeine disposition in human subjects. Herein, we report a method for the simultaneous quantification of caffeine and its three major metabolites in human plasma by high-performance liquid chromatography coupled to electrospray tandem mass spectrometry (HPLC-ESI-MS/MS). Human plasma samples were treated by simple protein precipitation and the analytes were separated using a 6 min gradient program. Precision and accuracy were well within in the 15% acceptance range. The simple sample preparation, short runtime, sensitivity and the inclusion of caffeine's major metabolites make this assay methodology optimal for the study of caffeine's pharmacokinetics and pharmacodynamics in human subjects. Copyright © 2016 John Wiley & Sons, Ltd.

  7. Health benefits of methylxanthines in neurodegenerative diseases.

    Science.gov (United States)

    Oñatibia-Astibia, Ainhoa; Franco, Rafael; Martínez-Pinilla, Eva

    2017-06-01

    Methylxanthines (MTXs) are consumed by almost everybody in almost every area of the world. Caffeine, theophylline and theobromine are the most well-known members of this family of compounds; they are present, inter alia, in coffee, tea, cacao, yerba mate and cola drinks. MTXs are readily absorbed in the gastrointestinal tract and are able to penetrate into the central nervous system, where they exert significant psychostimulant actions, which are more evident in acute intake. Coffee has been paradigmatic, as its use was forbidden in many diseases, however, this negative view has radically changed; evidence shows that MTXs display health benefits in diseases involving cell death in the nervous system. This paper reviews data that appraise the preventive and even therapeutic potential of MTXs in a variety of neurodegenerative diseases. Future perspectives include the use of MTXs to advance the understanding the pathophysiology of, inter alia, Alzheimer's disease (AD) and Parkinson's disease (PD), and the use of the methylxanthine chemical moiety as a basis for the development of new and more efficacious drugs. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Development of urine standard reference materials for metabolites of organic chemicals including polycyclic aromatic hydrocarbons, phthalates, phenols, parabens, and volatile organic compounds

    Science.gov (United States)

    Schantz, Michele M.; Benner, Bruce A.; Heckert, N. Alan; Sander, Lane C.; Sharpless, Katherine E.; Vander Pol, Stacy S.; Vasquez, Y.; Villegas, M.; Wise, Stephen A.; Alwis, K. Udeni; Blount, Benjamin C.; Calafat, Antonia M.; Li, Zheng; Silva, Manori J.; Ye, Xiaoyun; Gaudreau, Éric; Patterson, Donald G.; Sjödin, Andreas

    2016-01-01

    Two new Standard Reference Materials (SRMs), SRM 3672 Organic Contaminants in Smokers’ Urine (Frozen) and SRM 3673 Organic Contaminants in Non-Smokers’ Urine (Frozen), have been developed in support of studies for assessment of human exposure to select organic environmental contaminants. Collaborations among three organizations resulted in certified values for 11 hydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) and reference values for 11 phthalate metabolites, 8 environmental phenols and parabens, and 24 volatile organic compound (VOC) metabolites. Reference values are also available for creatinine and the free forms of caffeine, theobromine, ibuprofen, nicotine, cotinine, and 3-hydroxycotinine. These are the first urine Certified Reference Materials characterized for metabolites of organic environmental contaminants. Noteworthy, the mass fractions of the environmental organic contaminants in the two SRMs are within the ranges reported in population survey studies such as the National Health and Nutrition Examination Survey (NHANES) and the Canadian Health Measures Survey (CHMS). These SRMs will be useful as quality control samples for ensuring compatibility of results among population survey studies and will fill a void to assess the accuracy of analytical methods used in studies monitoring human exposure to these organic environmental contaminants. PMID:25651899

  9. Analysis of Camellia sinensis green and black teas via ultra high performance liquid chromatography assisted by liquid-liquid partition and two-dimensional liquid chromatography (size exclusion × reversed phase).

    Science.gov (United States)

    Scoparo, Camila T; de Souza, Lauro M; Dartora, Nessana; Sassaki, Guilherme L; Gorin, Philip A J; Iacomini, Marcello

    2012-01-27

    Green and black teas (Camellia sinensis) contain compounds ranging from simple phenolics to complex glycosides, many of which have well-recognized health benefits. Here, we describe two methodologies aiming to achieve a comprehensive analysis of hydro-alcoholic extracts of C. sinensis. In the first step, the extracts were partitioned in water, n-butanol, ethyl acetate and chloroform to separate the compounds according to their polarity, yielding less complex samples to be analyzed by ultra high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS). Additionally, a comprehensive two dimensional liquid chromatography (2D-LC) technique, employing size exclusion chromatography (SEC) × reversed phase (BEH-C18) was developed. The following compounds were identified on the basis of retention time, UV-spectra and MS fragmentation patterns: catechins, theaflavins and their gallate derivatives; kaempferol, quercetin and myricetin mono-, di-, tri- and tetraglycosides; esters of quinic acid and gallic or hydroxycinnamic acids; purine alkaloids, such as caffeine and theobromine and many lipids. Additionally, there were many novel compounds that were previously undescribed, such as saponin isomers and gallic acid esters of four glycosides of myricetin, quercetin and kaempferol. Copyright © 2011 Elsevier B.V. All rights reserved.

  10. Potency and Utilization of Cocoa Pod Husk as an Alternative Feed for Ruminants

    Directory of Open Access Journals (Sweden)

    Wisri Puastuti

    2014-09-01

    Full Text Available Cocoa pod husk (CPH is potential as an alternative source of feed for ruminants due to its quantity and quality. The availability of CPH in Indonesia during the harvest season is abundant and able to feed around 635,305 animal unit (AU per year. Cocoa pod husk as a feedstuff which contains crude protein between 6.80-13.78%; NDF 55.30-73.90% and ADF 38.31-58.98% is a source of fiber and can replace grass. However, CPH also contains antinutritional compounds such as lignin, tannin and theobromine. In order to optimize the use of CPH as feed, treatments are needed to improve the nutritive value and digestibility, to reduce negative effects and to extend shelf life. Processing methods can be conducted physically, chemically and microbiologically. Processing can improve the nutritional value of CPH especially the protein content. Feeding livestock using processed CPH showed better weight gain than that of unprocessed CPH. Besides, the attempt to utilize CPH as feed can overcome the shortage of forage, especially during dry season and increase ruminant productivity around the area of cocoa plantations.

  11. Evaluation of ice-tea quality by DART-TOF/MS.

    Science.gov (United States)

    Rajchl, Aleš; Prchalová, Jana; Kružík, Vojtěch; Ševčík, Rudolf; Čížková, Helena

    2015-11-01

    DART (Direct Analysis in Real Time) coupled with Time-of-Flight Mass Spectrometry (TOF/MS) has been used for analyses of ice-teas. The article focuses on quality and authenticity of ice-teas as one of the most important tea-based products on the market. Twenty-one samples of ice-teas (black and green) were analysed. Selected compounds of ice-teas were determined: theobromine, caffeine, total phenolic compounds, total soluble solids, total amino acid concentration, preservatives and saccharides were determined. Fingerprints of DART-TOF/MS spectra were used for comprehensive assessment of the ice-tea samples. The DART-TOF/MS method was used for monitoring the following compounds: citric acid, caffeine, saccharides, artificial sweeteners (saccharin, acesulphame K), and preservatives (sorbic and benzoic acid), phosphoric acid and phenolic compounds. The measured data were subjected to a principal components analysis. The HPLC and DART-TOF/MS methods were compared in terms of determination of selected compounds (caffeine, benzoic acid, sorbic acid and saccharides) in the ice-teas. The DART-TOF/MS technique seems to be a suitable method for fast screening, testing quality and authenticity of tea-based products. Copyright © 2015 John Wiley & Sons, Ltd.

  12. Achocolatados: análise química Chocolate drink powders: chemical analysis

    Directory of Open Access Journals (Sweden)

    Mércia de Freitas Eduardo

    2004-09-01

    Full Text Available Neste trabalho avaliaram-se propriedades químicas de achocolatados do mercado brasileiro, sendo estas os teores de lipídios, proteínas, cinzas, umidade, pH, teobromina e alcalóides totais. Estas propriedades são influenciadas principalmente pelo conteúdo de cacau em pó e pelo conteúdo dos ingredientes lácteos, como o leite em pó e o soro de leite em pó. Foi feito um comparativo entre as marcas de achocolatados, dando ênfase às grandes diferenças, quando existentes, entre os achocolatados dietéticos (para dietas de restrição de sacarose, frutose e glicose (dextrose e "light" (com redução de açúcares e os tradicionais.Some chemical properties of chocolate drink powders from Brazilian market, such as lipid, protein, ashes, moisture, pH, theobromine and total alkaloids content of the samples were evaluated. These properties are mainly influenced by cocoa powder content and by dairy products, such as milk powder and whey. A comparison among the brands, emphasizing their strong differences, when existing, among diet (without sugars and light (low sugar and the standard brands was made.

  13. New Sample Preparation Method for Quantification of Phenolic Compounds of Tea (Camellia sinensis L. Kuntze: A Polyphenol Rich Plant

    Directory of Open Access Journals (Sweden)

    P. A. Nimal Punyasiri

    2015-01-01

    Full Text Available Chemical analysis of the Sri Lankan tea (Camellia sinensis, L. germplasm would immensely contribute to the success of the tea breeding programme. However, the polyphenols, particularly catechins (flavan-3-ols, are readily prone to oxidation in the conventional method of sample preparation. Therefore, optimization of the present sample preparation methodology for the profiling of metabolites is much important. Two sample preparation methodologies were compared, fresh leaves (as in the conventional procedures and freeze-dried leaves (a new procedure, for quantification of major metabolites by employing two cultivars, one is known to be high quality black tea and the other low quality black tea. The amounts of major metabolites such as catechins, caffeine, gallic acid, and theobromine, recorded in the new sampling procedure via freeze-dried leaves, were significantly higher than those recorded in the conventional sample preparation procedure. Additionally new method required less amount of leaf sample for analysis of major metabolites and facilitates storage of samples until analysis. The freeze-dried method would be useful for high throughput analysis of large number of samples in shorter period without chemical deterioration starting from the point of harvest until usage. Hence, this method is more suitable for metabolite profiling of tea as well as other phenol rich plants.

  14. Xanthine-Catechin Mixture Enhances Lithium-Induced Anti-Inflammatory Response in Activated Macrophages In Vitro

    Directory of Open Access Journals (Sweden)

    Fernanda Barbisan

    2017-01-01

    Full Text Available Lithium (Li is a chemical element used for treating and preventing bipolar disorder (BD and exerts positive effects such as anti-inflammatory effects as well as undesirable side effects. These effects of Li can be influenced by interaction with some nutritional elements. Therefore, we investigated the potential effects of xanthine (caffeine and theobromine and catechin molecules present in some food beverages broadly consumed worldwide, such as coffee and tea, on Li-induced anti-inflammatory effects. In the present study, we concomitantly exposed RAW 264.7 macrophages to Li, isolated xanthine and catechin molecules, and a xanthine-catechin mixture (XC mixture. We evaluated the effects of these treatments on cell proliferation, cell cycle progression, oxidative and antioxidant marker expression, cytokine levels, gene expression, and GSK-3β enzyme expression. Treatment with the XC mixture potentialized Li-induced anti-inflammatory effects by intensification of the following: GSK-3β inhibitory action, lowering effect on proinflammatory cytokines (IL-1β, IL-6, and TNFα, and increase in the levels of IL-10 that is an anti-inflammatory cytokine. Despite the controversial nature of caffeine consumption by BD patients, these results suggested that consumption of caffeine, in low concentrations, mixed with other bioactive molecules along with Li may be safe.

  15. Lysozyme binding ability toward psychoactive stimulant drugs: Modulatory effect of colloidal metal nanoparticles.

    Science.gov (United States)

    Sonu, Vikash K; Islam, Mullah Muhaiminul; Rohman, Mostofa Ataur; Mitra, Sivaprasad

    2016-10-01

    The interaction and binding behavior of the well-known psychoactive stimulant drugs theophylline (THP) and theobromine (THB) with lysozyme (LYS) was monitored by in-vitro fluorescence titration and molecular docking calculations under physiological condition. The quenching of protein fluorescence on addition of the drugs is due to the formation of protein-drug complex in the ground state in both the cases. However, the binding interaction is almost three orders of magnitude stronger in THP, which involves mostly hydrogen bonding interaction in comparison with THB where hydrophobic binding plays the predominant role. The mechanism of fluorescence quenching (static type) remains same also in presence of gold and silver nanoparticles (NPs); however, the binding capacity of LYS with the drugs changes drastically in comparison with that in aqueous buffer medium. While the binding affinity of LYS to THB increases ca. 100 times in presence of both the NPs, it is seen to decrease drastically (by almost 1000 fold) for THP. This significant modulation in binding behavior indicates that the drug transportation capacity of LYS can be controlled significantly with the formation protein-NP noncovalent assembly system as an efficient delivery channel. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Enzyme functional evolution through improved catalysis of ancestrally nonpreferred substrates

    Science.gov (United States)

    Huang, Ruiqi; Hippauf, Frank; Rohrbeck, Diana; Haustein, Maria; Wenke, Katrin; Feike, Janie; Sorrelle, Noah; Piechulla, Birgit; Barkman, Todd J.

    2012-01-01

    In this study, we investigated the role for ancestral functional variation that may be selected upon to generate protein functional shifts using ancestral protein resurrection, statistical tests for positive selection, forward and reverse evolutionary genetics, and enzyme functional assays. Data are presented for three instances of protein functional change in the salicylic acid/benzoic acid/theobromine (SABATH) lineage of plant secondary metabolite-producing enzymes. In each case, we demonstrate that ancestral nonpreferred activities were improved upon in a daughter enzyme after gene duplication, and that these functional shifts were likely coincident with positive selection. Both forward and reverse mutagenesis studies validate the impact of one or a few sites toward increasing activity with ancestrally nonpreferred substrates. In one case, we document the occurrence of an evolutionary reversal of an active site residue that reversed enzyme properties. Furthermore, these studies show that functionally important amino acid replacements result in substrate discrimination as reflected in evolutionary changes in the specificity constant (kcat/KM) for competing substrates, even though adaptive substitutions may affect KM and kcat separately. In total, these results indicate that nonpreferred, or even latent, ancestral protein activities may be coopted at later times to become the primary or preferred protein activities. PMID:22315396

  17. Uric Acid or 1-Methyl Uric Acid in the Urinary Bladder Increases Serum Glucose, Insulin, True Triglyceride, and Total Cholesterol Levels in Wistar Rats

    Directory of Open Access Journals (Sweden)

    T. Balasubramanian

    2003-01-01

    Full Text Available In animals deprived of food for a long period, a drop in the fat mass below 5% of the total body mass results in an increase in blood glucocorticoids and uric acid levels, followed by foraging activity. Since the glucocorticoids increase the uric acid excretion, an increase in the level of uric acid in the bladder urine could be the signal for this feeding behaviour and subsequent fat storage. Accumulation of fat is associated with hyperglycaemia, hyperinsulinaemia, hyperlipidaemia, and hypercholesterolaemia as seen in the metabolic syndrome or hibernation. It is hypothesized that uric acid or its structurally related compound, 1-methyl uric acid (one of the metabolites of the methyl xanthines namely caffeine, theophylline, and theobromine present in coffee, tea, cocoa, and some drugs, can act on the urinary bladder mucosa and increases the blood glucose, insulin, triglyceride, and cholesterol levels. In rats, perfusion of the urinary bladder with saturated aqueous solution of uric acid or 1-methyl uric acid results in a significant increase in the serum levels of glucose, insulin, true triglyceride, and total cholesterol in comparison with perfusion of the bladder with distilled water at 20, 40, and 80 min. The uric acid or the 1-methyl uric acid acts on the urinary bladder mucosa and increases the serum glucose, insulin, true triglyceride, and total cholesterol levels.

  18. Quantitative Analysis of Major Constituents in Green Tea with Different Plucking Periods and Their Antioxidant Activity

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    Lan-Sook Lee

    2014-07-01

    Full Text Available The objective of this study was to determine the relationship between the plucking periods and the major constituents and the antioxidant activity in green tea. Green tea was prepared from leaves plucked from the end of April 2013 to the end of May 2013 at intervals of one week or longer. The contents of theanine, theobromine, caffeine, catechin (C, and gallocatechin gallate (GCg were significantly decreased, whereas those of epicatechin (EC, epigallocatechin gallate (EGCg and epigallocatechin (EGC were significantly increased along with the period of tea leaf plucking. In addition, antioxidant activity of green tea and standard catechins was investigated using ABTS, FRAP and DPPH assays. The highest antioxidant activity was observed in relatively the oldest leaf, regardless of the assay methods used. Additionally, the order of antioxidant activity of standard catechins was as follows: EGCg ³ GCg ³ ECg > EGC ³ GC ³ EC ³ C. Moreover, the cis-catechins contents were the key factor affecting the antioxidant activity of green tea in all assays employed (ABTS, r = 0.731, p < 0.01; FRAP, r = 0.886, p < 0.01; DPPH, r = 0.778, p < 0.01.

  19. ANTIPSYCHOTICS REVERSE P-GLYCOPROTEIN-MEDIATED DOXORUBICIN RESISTANCE IN HUMAN UTERINE SARCOMA MES-SA/Dx5 CELLS: A NOVEL APPROACH TO CANCER CHEMOTHERAPY.

    Science.gov (United States)

    Angelini, A; Ciofani, G; Conti, P

    2015-01-01

    Multidrug resistance (MDR) mediated by P-glycoprotein (Pgp) remains one of the major obstacles to effective cancer chemotherapy. Several chemosensitizers have been used in vivo and in vitro to reverse MDR but have exhibited several unwanted side effects. Antipsychotics are often administered to treat psychiatric disorders such as delirium, anxiety and sleep disorders in cancer patients during chemotherapy. The present in vitro study, examined the effects of two common antipsychotic compounds, haloperidol and risperidone, and a natural compound such as theobromine on reversing MDR Pgp-mediated, to evaluate their potential use as chemosensitizing agents. The human doxorubicin (doxo) resistant uterine sarcoma cells (MES-SA/Dx5) that overexpress Pgp (100-fold), were treated with the antipsychotic alone (1, 10 and 20 μM) or in combination with different concentrations of doxo (2, 4 and 8 μM). The accumulation and cytotoxicity of doxo (MTT assay) and cellular GSH content (GSH assay) in comparison with verapamil, a well-known Pgp inhibitor, used as reference molecule were examined. It was found that the three compounds significantly enhanced the intracellular accumulation of doxo in resistant cancer cells, when compared with cells receiving doxo alone (p 30%) in resistant cells, when compared to untreated control cells (ptheobromine showed to be an effective Pgp inhibitor with the lowest toxicity.

  20. Alkaloids and phenolics biosynthesis increases mango resistance to infection by Ceratocystis fimbriata

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    Leonardo Araujo

    2016-01-01

    Full Text Available ABSTRACT Mango wilt, caused by Ceratocystis fimbriata, is one of the most important diseases affecting mango yields in Brazil. Information regarding the biochemical mechanisms involved in mango resistance against C. fimbriata is absent in the literature. Thus, the present study determined and quantified alkaloids and phenolics in the stem tissue of mango plants from Palmer (susceptible and Ubá (resistant cultivars. Furthermore, it was examined the effect of these secondary metabolites against C. fimbriata growth in vitro. The high-performance liquid chromatography revealed that the concentration of two alkaloids (theobromine and 7-methylxanthine and six phenolic compounds (caffeic acid, p-coumaric acid, gallic acid, protocatechuic acid, catechin and epicatechin in the inoculated plants from cv. Ubá was higher in comparison with inoculated plants from cv. Palmer. The concentration of the secondary metabolites was higher in the non-inoculated plants from cv. Palmer than in the inoculated ones, while the opposite was observed for plants of cv. Ubá. Peaks in the concentrations of secondary metabolites in the inoculated plants from both cultivars occurred at 7 and 14 days after inoculation. The different concentrations (10 to 30 mg∙mL−1 of secondary metabolites added to the Petri dishes greatly inhibited C. fimbriata growth over time. These results suggest that secondary metabolites played an important role in the resistance of mango plants against C. fimbriata infection.

  1. In vitro antioxidant capacity of tea of Echinodorus grandiforus, “leather hat,” in Wistar rat liver

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    RAFAELA F. LUNARDI

    2014-09-01

    Full Text Available Oxidative stress has been considered as one of the factors responsible for hepatic diseases, which sometimes require new ways of treatment. The present study aimed to evaluate the in vitro antioxidant capacity of the tea of Echinodorus grandiforus (“leather hat” plant in rat liver. Different preparations of tea were evaluated for phenolic composition, antioxidant activity by DPPH assay and ability to inhibit lipid peroxidation induced by copper sulfate. The antioxidant activity was assessed in liver tissue treated with sodium azide in the presence or absence of tea by assays for lipid peroxidation (TBARS, protein oxidation (carbonyl and the antioxidant enzymes catalase (CAT and superoxide dismutase (SOD. The results show that different preparations of tea are important sources of polyphenols and contain theobromine, catechin and vitexin. Furthermore, the results indicate that this tea exhibits an antioxidant activity by its ability to scavenge DPPH radical. Different preparations of tea prevented damage to lipids and proteins induced by sodium azide, as well as assisting in restoring CAT and SOD activities. Thus, it can be seen that E. grandiforus tea had antioxidant activity in serum and liver being able to prevent oxidative damages generated by sodium azide.

  2. Phytochemical profile of morphologically selected yerba-mate progenies

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    Alice Teresa Valduga

    2016-02-01

    Full Text Available ABSTRACT Yerba-mate (Ilex paraguariensis St. Hil is a native South American species. Plant progenies are populations that differ in terms of their productivity, morphology and phytochemical profile. This study aimed to determine the concentration of primary and secondary metabolites, such as antioxidants, in leaves, of yerba-mate progenies selected based on morphological characteristics. We evaluated the centesimal composition of secondary metabolites in the leaves of five yerba-mate plants. Methylxanthines and phenolic compounds were determined by UPLC-PDA, and antioxidant activity by measuring DPPH scavenging. Significant differences were found in centesimal composition and the contents of caffeine, theobromine, rutin and chlorogenic acid, as well as antioxidant activities, in selected progenies. The IC50 values were correlated with the chlorogenic acid levels (r2 = 0.5242 and soluble content (r2 = 0.7686. The morphological characteristics observed in yerba-mate leaves can be used as a tool for plant selection, to obtain matrices with different phytochemical profiles as a genetic material source.

  3. Consumption of chocolate in pregnant women and risk of preeclampsia: a systematic review

    Science.gov (United States)

    2013-01-01

    Background Previous studies have been limited in reporting the association between chocolate consumption, measured by interviewer-administered questionnaire or serum theobromine, a biomarker for cocoa, and risk of preeclampsia, and have showed somewhat conflicting results. Methods/Design A systematic review of observational and experimental studies will be carried out. We will examine PubMed, Embase, and the entire Cochrane Library. Studies of chocolate consumption compared or not with placebo or low flavanol chocolate during pregnancy will be evaluated to investigate the effect of chocolate consumption in pregnant women on the risk of preeclampsia or pregnancy-induced hypertension. Screening for inclusion, data extraction, and quality assessment will be performed independently by two reviewers in consultation with a third reviewer. Validity of the studies will be ascertained by using the Cochrane Collaboration’s tool. Relative risk of preeclampsia will be the primary measure of treatment effect. Heterogeneity will be explored by subgroup analysis according to confounding factors and bias. Discussion This systematic review will contribute to establish the current state of knowledge concerning the possible association between chocolate consumption and prevention of preeclampsia. Furthermore, it will justify if additional experimental trials are necessary to better evaluate the benefits of chocolate consumption on the risk of preeclampsia. Trial registration This systematic review has been registered in the PROSPERO international prospective register of systematic reviews. The registration number is: CRD42013005338 PMID:24360219

  4. Chemical Composition and Hypotensive Effect of Campomanesia xanthocarpa

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    Liane Santariano Sant’Anna

    2017-01-01

    Full Text Available Campomanesia xanthocarpa is known in Brazil as Guabiroba and is popularly used for various diseases, such as inflammatory, renal, and digestive diseases and dyslipidemia. The aim of the study was to analyze the chemical composition and investigate the effects of aqueous extract of C. xanthocarpa on the blood pressure of normotensive rats, analyzing the possible action mechanism using experimental and in silico procedures. The extract was evaluated for total phenolic compounds and total flavonoid content. The chemical components were determined by HPLC analyses. Systolic and diastolic blood pressure and heart rate were measured with extract and drugs administration. The leaves of C. xanthocarpa presented the relevant content of phenolics and flavonoids, and we suggested the presence of chlorogenic acid, gallic acid, quercetin, and theobromine. The acute administration of aqueous extract of C. xanthocarpa has a dose-dependent hypotensive effect in normotensive rats, suggesting that the action mechanism may be mediated through the renin-angiotensin system by AT1 receptor blockade and sympathetic autonomic response. Docking studies showed models that indicated an interaction between chlorogenic acid and quercetin with the AT1 receptor (AT1R active site. The findings of these docking studies suggest the potential of C. xanthocarpa constituents for use as preventive agents for blood pressure.

  5. Modification of radiation-induced division delay by caffeine analogues and dibutyryl cyclic AMP

    Energy Technology Data Exchange (ETDEWEB)

    Kimler, B.F.; Leeper, D.B.; Snyder, M.H.; Rowley, R.; Schneiderman, M.H. (Thomas Jefferson Univ., Philadelphia, PA (USA). Hospital)

    1982-01-01

    The mitotic selection procedure for cell cycle analysis was utilized to investigate the concentration-dependent modification of x-radiation-induced division delay in Chinese hamster ovary (CHO) cells by methyl xanthines (caffeine, theophylline, and theobromine) and by dibutyryl cyclic AMP. The methyl xanthines (concentrations from 0.5 to 1000 ..mu..g/ml) all reduced radiation-induced division delay with the effect being linear between approximately 100 and 1000 ..mu..g/ml. After doses of 100-300 rad, delay was reduced by 75, 94 or 83 per cent at 1000 ..mu..g/ml for each drug, respectively. However, the addition of dibutyryl cyclic AMP had an opposite effect: radiation-induced delay was increased by the concentration range of 0.3 to 300 ..mu..g/ml. These results indicate that in mammalian cells the control of cell cycle progression and the modification of radiation-induced division delay are not simply related to intracellular levels of cyclic AMP. Rather, there appear to be at least two competing mechanisms which are differentially affected by caffeine analogues or by direct addition of dibutyryl cyclic AMP. The direct effect of caffeine and the methyl xanthines on membrane calcium permeability is considered.

  6. “RaMassays”: Synergistic Enhancement of Plasmon-Free Raman Scattering and Mass Spectrometry for Multimodal Analysis of Small Molecules

    Science.gov (United States)

    Alessandri, Ivano; Vassalini, Irene; Bertuzzi, Michela; Bontempi, Nicolò; Memo, Maurizio; Gianoncelli, Alessandra

    2016-10-01

    SiO2/TiO2 core/shell (T-rex) beads were exploited as “all-in-one” building-block materials to create analytical assays that combine plasmon-free surface enhanced Raman scattering (SERS) and surface assisted laser desorption/ionization (SALDI) mass spectrometry (RaMassays). Such a multi-modal approach relies on the unique optical properties of T-rex beads, which are able to harvest and manage light in both UV and Vis range, making ionization and Raman scattering more efficient. RaMassays were successfully applied to the detection of small (molecular weight, M.W. theobromine couples demonstrated the synergistic reciprocal reinforcement of SERS and SALDI. Finally, the conversion of L-tyrosine in L-DOPA was utilized to probe RaMassays as analytical tools for characterizing reaction intermediates without introducing any spurious effects. RaMassays exhibit important advantages over plasmonic nanoparticles in terms of reproducibility, absence of interference and potential integration in multiplexed devices.

  7. Studies on the biotechnological production of methyl xanthines. Untersuchung zur biotechnologischen Gewinnung von Methylxanthinen

    Energy Technology Data Exchange (ETDEWEB)

    Glueck, M.

    1986-04-25

    Two strains of Pseudomonas putida were isolated during enrichment of caffeine-degrading microorganisms of the two, the strain Pseudomonas putida WS grew on media containing up to 2% of caffeine as the sole source of carbon and nitrogen. Of 25 further strains of bacteria and fungi, only one yeast also had caffeine-degrading characteristics. Caffeine degradation was much slower than with the two Pseudomonas strains, and caffeine was used only as a nitrogen source. This suggests two alternative methods of methyl xanthine production: 1) When graining Mutant H8 on glucose caffeine, the reaction could be interrupted with good yields after a short time. This method requires the processing of a complex mixture of caffeine, theobromine, and heteroxanthine. 2) When growing Mutant H8 on a mixture of peptone meat broth plus caffeine, the reaction can be interrupted with average yields after near-complete conversion. This method is recommended for heteroxanthine production, owing to the fact that peak heteroxanthine yields of 50% are not reached until all other methyl anthines have been converted more or less completely.

  8. Structure-Bioactivity Relationships of Methylxanthines: Trying to Make Sense of All the Promises and the Drawbacks.

    Science.gov (United States)

    Monteiro, João P; Alves, Marco G; Oliveira, Pedro F; Silva, Branca M

    2016-07-27

    Methylxanthines are a group of phytochemicals derived from the purine base xanthine and obtained from plant secondary metabolism. They are unobtrusively included in daily diet in common products as coffee, tea, energetic drinks, or chocolate. Caffeine is by far the most studied methylxanthine either in animal or epidemiologic studies. Theophylline and theobromine are other relevant methylxanthines also commonly available in the aforementioned sources. There are many disseminated myths about methylxanthines but there is increased scientific knowledge to discuss all the controversy and promise shown by these intriguing phytochemicals. In fact, many beneficial physiologic outcomes have been suggested for methylxanthines in areas as important and diverse as neurodegenerative and respiratory diseases, diabetes or cancer. However, there have always been toxicity concerns with methylxanthine (over)consumption and pharmacologic applications. Herein, we explore the structure-bioactivity relationships to bring light those enumerated effects. The potential shown by methylxanthines in such a wide range of conditions should substantiate many other scientific endeavors that may highlight their adequacy as adjuvant therapy agents and may contribute to the advent of functional foods. Newly designed targeted molecules based on methylxanthine structure may originate more specific and effective outcomes.

  9. Nutritional composition, bioactive compounds and volatile profile of cocoa beans from different regions of Cameroon.

    Science.gov (United States)

    Caprioli, Giovanni; Fiorini, Dennis; Maggi, Filippo; Nicoletti, Marcello; Ricciutelli, Massimo; Toniolo, Chiara; Prosper, Biapa; Vittori, Sauro; Sagratini, Gianni

    2016-06-01

    Analysis of the complex composition of cocoa beans provides fundamental information for evaluating the quality and nutritional aspects of cocoa-based food products, nutraceuticals and supplements. Cameroon, the world's fourth largest producer of cocoa, has been defined as "Africa in miniature" because of the variety it habitats. In order to evaluate the nutritional characteristics of cocoa beans from five different regions of Cameroon, we studied their polyphenolic content, volatile compounds and fatty acids composition. The High Performance Thin Layer Chromatography (HPTLC) analysis showed that the Mbalmayo sample had the highest content of theobromine (11.6 mg/g) and caffeic acid (2.1 mg/g), while the Sanchou sample had the highest level of (-)-epicatechin (142.9 mg/g). Concerning fatty acids, the lowest level of stearic acid was found in the Mbalmayo sample while the Bertoua sample showed the highest content of oleic acid. Thus, we confirmed that geographical origin influences the quality and nutritional characteristics of cocoa from these regions of Cameroon.

  10. Interactions between minimum run time, modifier concentration, and efficiency parameters in a high performance liquid chromatography separation.

    Science.gov (United States)

    Chester, T L; Stalcup, A M

    2011-01-14

    We modeled and studied the separation of uracil, nicotinamide, resorcinol, theobromine, theophylline, and caffeine on four C-18 columns of different lengths packed with the same stationary phase using water/methanol mobile phase at one temperature. Predictions of retention times and peak widths were compared with experimental results and were found to be sufficiently accurate for performing optimization calculations. With limits set on the required resolution and on maximum values for pressure and flow rate, calculations were performed for numerous virtual column lengths seeking the smallest possible analysis time for each length while allowing methanol concentration and flow rate to vary as required to minimize run time. Predictions were experimentally verified for the column lengths actually available. These calculations revealed the dependence of best-possible analysis time on column length, modifier concentration, flow rate, and pressure for the real system that was modeled, and provided insight into parameter interactions with respect to analysis times meeting the needs and limits specified. We show that when these parameters are considered in concert, rather than individually, conventional guidelines regarding setting their values may not always lead to the optimum. Copyright © 2010 Elsevier B.V. All rights reserved.

  11. Use of pharmacological agents to implicate a role for phosphoinositide hydrolysis products in malaria gamete formation.

    Science.gov (United States)

    Ogwan'g, R; Mwangi, J; Gachihi, G; Nwachukwu, A; Roberts, C R; Martin, S K

    1993-11-02

    The kinetics of phosphoinositol 4,5 bisphosphate hydrolysis products in activated Plasmodium falciparum gametocytes suggests a role for inositol trisphosphate [Ins(1,4,5)P3] and diacylglycerol (DAG) in the signal transduction pathway of malaria gametocytes. To investigate further this role, compounds that have an effect on the metabolism and biologic functions of these second messengers were tested in an in vitro system. Gentamycin, 2,3 diphosphoglycerate (2,3 DPG) and magnesium ion (Mg2+), inhibitors of Ins(1,4,5)P3 5' phosphatase, all stimulated gametocytes to exflagellate in suspended animation buffer, pH 7.4, at room temperature. In addition, methylxanthines, caffeine and theobromine, calcium ionophore (A-23187), and external calcium also stimulated exflagellation. In contrast, neomycin, an aminoglycoside that inhibits phospholipase C activity, and heparin, an antagonist of Ins(1,4,5)P3 binding to its receptor, inhibited microgamete formation. Quinine and chloroquine which can inhibit both phospholipase A and C activity also inhibited gametocyte exflagellation. The consistent manner in which these various compounds affect gametocyte activation further implicates phosphoinositol turnover in the signal transduction pathway of falciparum gametocytes.

  12. HPLC analysis of methylxanthines in human breast milk.

    Science.gov (United States)

    Blanchard, J; Weber, C W; Shearer, L E

    1990-12-01

    A sensitive and specific high-performance liquid chromatographic (HPLC) procedure is developed for simultaneously quantitating the levels of caffeine, theophylline, theobromine and paraxanthine in breast milk. The method involved the precipitation of proteins present in the milk samples with a 6% v/v perchloric acid solution containing the internal standard, proxyphylline, followed by centrifugation at 12,800 Xg for 10 minutes. The clear supernatant was then chromatographed on a C18 reversed-phase analytical column at ambient temperature using a wavelength of 272 nm. Samples were eluted from the column at a constant flow rate of 1.5 mL/min using a gradient program in which the concentration of methanol in the mobile phase varied from 0 to 16%. The mean recoveries of the methylxanthines averaged over all the concentrations examined were generally excellent and ranged from 96.3 +/- 5.4% for caffeine to 102.3 +/- 8.9% for paraxanthine. The assay precision was very good and the peaks of interest were extremely well resolved. The method is recommended for assessing the total caffeine and dimethylxanthine load to which the nursing infant is exposed in mothers ingesting typical amounts of caffeine.

  13. Inelastic neutron scattering study of methyl groups rotation in some methylxanthines

    Science.gov (United States)

    Prager, M.; Pawlukojc, A.; Wischnewski, A.; Wuttke, J.

    2007-12-01

    The three isomeric dimethylxanthines and trimethylxanthine are studied by neutron spectroscopy up to energy transfers of 100meV at energy resolutions ranging from 0.7μeV to some meV. The loss of elastic intensity with increasing temperature can be modeled by quasielastic methyl rotation. The number of inequivalent methyl groups is in agreement with those of the room temperature crystal structures. Activation energies are obtained. In the case of theophylline, a doublet tunneling band is observed at 15.1 and 17.5μeV. In theobromine, a single tunneling band at 0.3μeV is found. Orientational disorder in caffeine leads to a 2.7μeV broad distribution of tunneling bands around the elastic line. At the same time, broad low energy phonon spectra characterize an orientational glassy state with weak methyl rotational potentials. Librational energies of the dimethylxanthines are clearly seen in the phonon densities of states. Rotational potentials can be derived which explain consistently all observables. While their symmetry in general is threefold, theophylline shows a close to sixfold potential reflecting a mirror symmetry.

  14. LC-DAD-ESI-MS/MS characterization of phenolic constituents in Turkish black tea: Effect of infusion time and temperature.

    Science.gov (United States)

    Kelebek, Hasim

    2016-08-01

    The influence of different infusion times and temperatures on phenolic, antioxidant, and color properties of black teas (Camellia sinensis L.) was investigated in the present study. The teas were prepared using infusion times of 3, 6, and 10min at 80 and 100°C. Liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MS/MS), whereby the two purine alkaloids were detected using positive ESI, and the other 33 phenolic compounds were detected using negative ESI. A total of 35 phenolic compounds were identified and quantified in the tea infusions, with the most abundant being gallic acid, (-)-epicatechin-gallate, 4-p-coumaroylquinic acid, quercetin-3-O-galactosyl-rhamnosyl-glucoside, kaempferol-3-O-glucosyl-rhamnosyl-glucoside, theaflavin, and theobromine. The antioxidant capacity of the tea infusions was determined using two different methods: DPPH (2,2-diphenyl-1-picryl hydrazyl) and ABTS (2,2'-azino-bis-3-ethyl-benzothiazoline-6-sulfonic acid) assays. DPPH and the ABTS values varied largely among the tea preparations. The increment in infusion times and temperature led to increase in the tea's antioxidant capacities. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Computationally-assisted approach to the vibrational spectra of molecular crystals: study of hydrogen-bonding and pseudo-polymorphism.

    Science.gov (United States)

    Nolasco, Mariela M; Amado, Ana M; Ribeiro-Claro, Paulo J A

    2006-10-13

    A new computationally-assisted methodology (PiMM), which accounts for the effects of intermolecular interactions in the crystal, is applied to the complete assignment of the Raman and infrared vibrational spectra of room temperature forms of crystalline caffeine, theobromine, and theophylline. The vibrational shifts due to crystal packing interactions are evaluated from ab initio calculations for a set of suitable molecular pairs, using the B3LYP/6-31G* approach. The proposed methodology provides an answer to the current demand for a reliable assignment of the vibrational spectra of these methyl-xanthines, and clarifies several misleading assignments. The most relevant intermolecular interactions in each system and their effect on the vibrational spectra are considered and discussed. Based on these results, significant insights are obtained for the structure of caffeine in the anhydrous form (stable at room temperature), for which no X-ray structure has been reported. A possible structure based on C((8))--H...N((9)) and C((1,3))--H...O intermolecular interactions is suggested.

  16. Determination of Flavonoids, Phenolic Acids, and Xanthines in Mate Tea (Ilex paraguariensis St.-Hil.

    Directory of Open Access Journals (Sweden)

    Mirza Bojić

    2013-01-01

    Full Text Available Raw material, different formulations of foods, and dietary supplements of mate demands control of the content of bioactive substances for which high performance thin layer chromatography (TLC, described here, presents simple and rapid approach for detections as well as quantification. Using TLC densitometry, the following bioactive compounds were identified and quantified: chlorogenic acid (2.1 mg/g, caffeic acid (1.5 mg/g, rutin (5.2 mg/g, quercetin (2.2 mg/g, and kaempferol (4.5 mg/g. The results obtained with TLC densitometry for caffeine (5.4 mg/g and theobromine (2.7 mg/g show no statistical difference to the content of total xanthines (7.6 mg/g obtained by UV-Vis spectrophotometry. Thus, TLC remains a technique of choice for simple and rapid analysis of great number of samples as well as a primary screening technique in plant analysis.

  17. Spontaneously Assembled Nano-aggregates in Clear Green Tea Infusions from Camellia ptilophylla and Camellia sinensis.

    Science.gov (United States)

    Lin, Xiaorong; Gao, Xiong; Chen, Zhongzheng; Zhang, Yuanyuan; Luo, Wei; Li, Xiaofei; Li, Bin

    2017-05-10

    Tea nano-aggregates spontaneously assembled in clear tea infusions are considered as the precursors of tea cream, although their molecular basis remains obscure. Here, we characterized nano-aggregates in green tea infusions from Camellia ptilophylla, a peculiar tea variety with 6.0% of theobromine, and Camellia sinensis as the control for comparative purpose. Numerous negatively charged spherical colloidal particles of 50-100 nm in diameter were primarily found in both green tea infusions. Catechins, proteins, and carbohydrates were confirmed as the dominant components in green tea nano-aggregates. In addition, iron, copper, nickel, proteins, and gallated catechins exhibited higher aggregating affinity than other components, whereas methylxanthines and calcium contributed to the transformation of nano-aggregates into tea cream. Green tea nano-aggregates were partly destroyed by simulated gastrointestinal digestion, and removing theses peculiar particles dramatically attenuated the bioaccessibility of methylxanthines, theanine, and some catechin monomers in green tea infusions. This study enhanced our knowledge of molecular interactions in the formation of green tea cream and provided insight into physicochemical profiles, phytochemical nature, and functional effects of green tea nano-aggregates.

  18. Molecularly imprinted poly(4-amino-5-hydroxy-2,7-naphthalenedisulfonic acid) modified glassy carbon electrode as an electrochemical theophylline sensor.

    Science.gov (United States)

    Aswini, K K; Vinu Mohan, A M; Biju, V M

    2016-08-01

    Theophylline is an inexpensive drug employed in asthma and chronic obstructive pulmonary disorder medications and is toxic at higher concentration. The development of a molecularly imprinted polymer based theophylline electrochemical sensor on glassy carbon electrode by the electropolymerization of 4-amino-5-hydroxy-2,7-naphthalenedisulfonic acid is being discussed in this work. The MIP modification enhances the theophylline recognition ability and the electron transfer kinetics of the bare electrode. The parameters, controlling the performance of the imprinted polymer based sensor, like number of electropolymerization cycles, composition of the pre-polymerization mixture, pH and immersion time were investigated and optimized. The interaction energy and the most stable conformation of the template-monomer complex in the pre-polymerization mixture were determined computationally using ab initio calculations based on density functional theory. The amperometric measurements showed that the developed sensor has a method detection limit of 0.32μM for the dynamic range of 0.4 to 17μM, at optimized conditions. The transducer possesses appreciable selectivity in the presence of structurally similar interferents such as theobromine, caffeine and doxofylline. The developed sensor showed remarkable stability and reproducibility and was also successfully employed in theophylline detection from commercially available tablets. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Modulation of radiation-induced apoptosis and G{sub 2}/M block in murine T-lymphoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Palayoor, S.T.; Macklis, R.M.; Bump, E.A.; Coleman, C.N. [Harvard Medical School, Boston, MA (United States)

    1995-03-01

    Radiation-induced apoptosis in lymphocyte-derived cell lines is characterized by endonucleolytic cleavage of cellular DNA within hours after radiation exposure. We have studied this phenomenon qualitatively (DNA gel electrophoresis) and quantitatively (diphenylamine reagent assay) in murine EL4 T-lymphoma cells exposed to {sup 137}Cs {gamma} irradiation. Fragmentation was discernible within 18-24 h after exposure. It increased with time and dose and reached a plateau after 8 Gy of {gamma} radiation. We studied the effect of several pharmacological agents on the radiation-induced G{sub 2}/M block and DNA fragmentation. The agents which reduced the radiation-induced G{sub 2}/M-phase arrest (caffeine, theobromine, theophylline and 2-aminopurine) enhanced the degree of DNA fragmentation at 24 h. In contrast, the agents which sustained the radiation-induced G{sub 2}/M-phase arrest (TPA, DBcAMP, IBMX and 3-aminobenzamide) inhibited the DNA fragmentation at 24 h. These studies on EL4 lymphoma cells are consistent with the hypothesis that cells with radiation-induced genetic damage are eliminated by apoptosis subsequent to a G{sub 2}/M block. Furthermore, it may be possible to modulate the process of radiation-induced apoptosis in lymphoma cells with pharmacological agents that modify the radiation-induced G{sub 2}/M block, and to use this effect in the treatment of patients with malignant disease. 59 refs., 7 figs.

  20. A novel electrochemical method to detect theophylline utilizing silver ions captured within abasic site-incorporated duplex DNA.

    Science.gov (United States)

    Ahn, Jun Ki; Park, Ki Soo; Won, Byoung Yeon; Park, Hyun Gyu

    2015-05-15

    We herein describe a novel and label-free electrochemical system to detect theophylline. The system was constructed by immobilizing duplex DNA containing an abasic site opposite cytosine on the gold electrode surface. In the absence of theophylline in a sample, silver ions freely bind to the empty abasic site in the duplex DNA leading to the highly elevated electrochemical signal by the redox reaction of silver ions. On the other hand, when theophylline is present, it binds to the abasic site by pseudo base pairing with the opposite cytosine nucleobase, which consequently prevents silver ions from binding to the abasic site. As a result, redox reaction of silver ions would be greatly reduced resulting in the accordingly decreased electrochemical signal. By employing this electrochemical strategy, theophylline was reliably detected at a concentration as low as 3.2 μM with the high selectivity over structurally similar substances such as caffeine and theobromine. Finally, the diagnostic capability of this method was also successfully verified by reliably detecting theophylline present in a real human serum sample with an excellent recovery ratio within 100±6%. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Alkaloids in the human food chain--natural occurrence and possible adverse effects.

    Science.gov (United States)

    Koleva, Irina I; van Beek, Teris A; Soffers, Ans E M F; Dusemund, Birgit; Rietjens, Ivonne M C M

    2012-01-01

    Alkaloid-containing plants are an intrinsic part of the regular Western diet. The present paper summarizes the occurrence of alkaloids in the food chain, their mode of action and possible adverse effects including a safety assessment. Pyrrolizidine alkaloids are a reason for concern because of their bioactivation to reactive alkylating intermediates. Several quinolizidine alkaloids, β-carboline alkaloids, ergot alkaloids and steroid alkaloids are active without bioactivation and mostly act as neurotoxins. Regulatory agencies are aware of the risks and have taken or are considering appropriate regulatory actions for most alkaloids. These vary from setting limits for the presence of a compound in feed, foods and beverages, trying to define safe upper limits, advising on a strategy aiming at restrictions in use, informing the public to be cautious or taking specific plant varieties from the market. For some alkaloids known to be present in the modern food chain, e.g., piperine, nicotine, theobromine, theophylline and tropane alkaloids risks coming from the human food chain are considered to be low if not negligible. Remarkably, for many alkaloids that are known constituents of the modern food chain and of possible concern, tolerable daily intake values have so far not been defined. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida L Catabolismo de cafeína e purificação de xantina oxidase responsável pela produção de ácidos metilúricos em Pseudomonas putida L

    Directory of Open Access Journals (Sweden)

    Dirce Mithico Yamaoka-Yano

    1999-01-01

    Full Text Available Caffeine catabolism and a xanthine oxidase involved in the alkaloid breakdown were studied in Pseudomonas putida L, a strain displaying high ability to grow on this substrate. Cells cultured with unlabelled caffeine and 14C labeled caffeine and xanthine showed that this alkaloid was broken-down via theobromine/paraxanthine -> 7-methylxanthine -> xanthine -> uric acid -> allantoin -> allantoic acid. Methyluric acids were formed from the oxidation of theobromine, paraxanthine and 7-methylxanthine, although no bacterial growth was observed on these compounds, indicating that this might be due to a wide substrate specificity of xanthine oxidase. This was confirmed by activity staining in PAGE where activity was observed with theophylline and 3-methylxanthine, which are not involved in the alkaloid breakdown. A single band of activity was detected without addition of NAD+, showing an oxidase form of the enzyme. The enzyme optimum temperature and pH were 30oC and 7.0, respectively. The determined Km was 169 µM, and the pI 3.1 - 4.0. The molecular weight determined by side by side comparison of activity staining of the enzyme in PAGE and PAGE of BSA was 192 kDa, which was coincident with the sum (198.4 kDa of three subunits (71, 65.6 and 61.8 kDa of the purified protein.O catabolismo de cafeína e a enzima xantina oxidase, envolvida na sua degradação, foram estudados em Pseudomonas putida L, uma linhagem com alta capacidade para utilizar este substrato como fonte de energia. Células crescidas na presença de cafeína e xantina marcadas com 14C, e cafeína não marcada, mostraram que este alcalóide foi degradado via teobromina/paraxantina -> 7-metilxantina -> xantina -> ácido úrico -> alantoína -> ácido alantóico. Ácidos metilúricos foram formados a partir de teobromina, paraxantina e 7-metilxantina, embora nenhum crescimento bacteriano ter sido observado quando estes compostos foram usados como substratos, indicando que a xantina oxidase

  3. Quantitative genetic analysis of methylxanthines and phenolic compounds in mate progenies Análise genética quantitativa de metilxantinas e compostos fenólicos em progênies de erva-mate

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    Euclides Lara Cardozo Junior

    2010-02-01

    Full Text Available The objective of this work was to determine the contents of methylxanthines, caffeine and theobromine, and phenolic compounds, chlorogenic and caffeic acids, in 51 mate progenies (half-sib families and estimate the heritability of genetic parameters. Mate progenies were from five Brazilian municipalities: Pinhão, Ivaí, Barão de Cotegipe, Quedas do Iguaçu, and Cascavel. The progenies were grown in the Ivaí locality. The contents of the compounds were obtained by high performance liquid chromatography (HPLC. The estimation of genetic parameters by the restricted maximum likelihood (REML and the prediction of genotypic values via best linear unbiased prediction (BLUP were obtained by the Selegen - REML/BLUP software. Caffeine (0.248-1.663% and theobromine (0.106-0.807% contents were significantly different (p0.5. The two different progeny groups determined for chlorogenic (1.365-2.281% and caffeic (0.027-0.037% acid contents were not significantly different (pO objetivo deste trabalho foi determinar o teor de metilxantinas, cafeína e teobromina, e de compostos fenólicos, ácido clorogênico e ácido cafeico, em 51 progênies de erva-mate e estimar componentes de variância e herdabilidade. As progênies de erva-mate eram oriundas de cinco municipios brasileiros: Pinhão, Ivaí, Barão do Cotegipe, Quedas do Iguaçu e Cascavel. Essas progênies foram cultivadas na localidade de Ivaí. O conteúdo dos compostos foi obtido por cromatografia líquida de alta eficiência (CLAE. Na estimativa dos componentes da variância e dos parâmetros genotípicos, utilizou-se a metodologia de modelos mistos para a obtenção da melhor predição linear não viciada (BLUP dos efeitos genotípicos e o processo da máxima verossimilhança restrita (REML, processados pelo programa Selegen - REML/BLUP. Os conteúdos de cafeína (0,248-1,663 % e teobromina (0,106-0.807% foram significativamente (p0,5. Foram determinados dois diferentes grupos de progênie para

  4. The remineralization potential of cocoa (Theobroma cacao bean extract to increase the enamel micro hardness

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    Sulistianingsih Sulistianingsih

    2017-08-01

    Full Text Available Introduction: Remineralization is the process of returning mineral ions into a hydroxyapatite structure characterized by mineral deposition on the enamel surface. The presence of mineral deposition would affect the micro hardness of tooth enamel. The use of fluorine as remineralization agent with side effects such as fluorosis. Cocoa bean extract contains theobromin that can be used as an alternative remineralization ingredients. The objectives was to determine micro hardness email after remineralization using cocoa bean extract as natural material and to compare with fluorine use as synthetic material. Methods: Thirty-six maxillary first premolar tooth crown was cut and planted in the epoxy resin. Teeth were then immersed in demineralization solution at pH 4 for 6 hours. The sample were divided into 2 groups, 18 for the fluorine group and the remaining group of cocoa extract. Vickers microhardness test was used before treatment, after demineralized and after remineralization. Results: Enamel microhardness value before treatment in the fluorine group average value was 376.17 VHN and the cocoa extract group was 357.33 VHN. After demineralization in fluorine group was 268,13 VHN and cocoa extract group was 235,93 VHN. After remineralization in fluorine group was 321,08 VHN and cocoa extract group was 293,86 VHN. The results of the analysis showed that the level of micro hardness email after remineralization was not significantly different in two groups (p > 0.05. Conclusions: Cocoa extract is able to increase the microhardness of enamel so it can act as a substitution for fluorine remineralization.

  5. Theobroma cacao increases cells viability and reduces IL-6 and sVCAM-1 level in endothelial cells induced by plasma from preeclamptic patients.

    Science.gov (United States)

    Rahayu, Budi; Baktiyani, Siti Candra Windu; Nurdiana, Nurdiana

    2016-01-01

    This study aims to investigate whether an ethanolic extract of Theobroma cacao bean is able to increase cell viability and decrease IL-6 and sVCAM-1 in endothelial cells induced by plasma from preeclamptic patients. Endothelial cells were obtained from human umbilical vascular endothelial cells. At confluency, endothelial cells were divided into six groups, which included control (untreated), endothelial cells exposed to plasma from normal pregnancy, endothelial cells exposed to 2% plasma from preeclamptic patients (PP), endothelial cells exposed to PP in the presence of ethanolic extract of T. cacao (PP+TC) at the following three doses: 25, 50, and 100 ppm. The analysis was performed in silico using the Hex 8.0, LigPlus and LigandScout 3.1 software. Analysis on IL-6 and sVCAM-1 levels were done by enzyme linked immunosorbent assay (ELISA). We found that seven of them could bind to the protein NFκB (catechin, leucoanthocyanidin, niacin, phenylethylamine, theobromine, theophylline, and thiamin). This increase in IL-6 was significantly (Pcacao extract. Plasma from PP significantly increased sVCAM-1 levels compared to untreated cells. This increase in sVCAM-1 was significantly attenuated by all doses of the extract. In conclusion, T. cacao extract prohibits the increase in IL-6 and sVCAM-1 in endothelial cells induced by plasma from preeclamptic patients. Therefore this may provide a herbal therapy for attenuating the endothelial dysfunction found in preeclampsia. Copyright © 2016 International Society for the Study of Hypertension in Pregnancy. Published by Elsevier B.V. All rights reserved.

  6. Diversity of cacao trees in Waslala, Nicaragua: associations between genotype spectra, product quality and yield potential.

    Science.gov (United States)

    Trognitz, Bodo; Cros, Emile; Assemat, Sophie; Davrieux, Fabrice; Forestier-Chiron, Nelly; Ayestas, Eusebio; Kuant, Aldo; Scheldeman, Xavier; Hermann, Michael

    2013-01-01

    The sensory quality and the contents of quality-determining chemical compounds in unfermented and fermented cocoa from 100 cacao trees (individual genotypes) representing groups of nine genotype spectra (GG), grown at smallholder plantings in the municipality of Waslala, Nicaragua, were evaluated for two successive harvest periods. Cocoa samples were fermented using a technique mimicking recommended on-farm practices. The sensory cocoa quality was assessed by experienced tasters, and seven major chemical taste compounds were quantified by near infrared spectrometry (NIRS). The association of the nine, partially admixed, genotype spectra with the analytical and sensory quality parameters was tested. The individual parameters were analyzed as a function of the factors GG and harvest (including the date of fermentation), individual trees within a single GG were used as replications. In fermented cocoa, significant GG-specific differences were observed for methylxanthines, theobromine-to-caffeine (T/C) ratio, total fat, procyanidin B5 and epicatechin, as well as the sensory attributes global score, astringency, and dry fruit aroma, but differences related to harvest were also apparent. The potential cocoa yield was also highly determined by the individual GG, although there was significant tree-to-tree variation within every single GG. Non-fermented samples showed large harvest-to-harvest variation of their chemical composition, while differences between GG were insignificant. These results suggest that selection by the genetic background, represented here by groups of partially admixed genotype spectra, would be a useful strategy toward enhancing quality and yield of cocoa in Nicaragua. Selection by the GG within the local, genetically segregating populations of seed-propagated cacao, followed by clonal propagation of best-performing individuals of the selected GG could be a viable alternative to traditional propagation of cacao by seed from open pollination. Fast and

  7. Influence of Echinacea purpurea intake during pregnancy on fetal growth and tissue angiogenic activity.

    Directory of Open Access Journals (Sweden)

    Joanna Chorostowska-Wynimko

    2008-04-01

    Full Text Available The process of angiogenesis and control of blood vessels sprouting are fundamental to human health, as they play key roles in many physiological and pathological conditions. Intake of different pharmaceuticals with antiangiogenic activity by pregnant women may lead to severe developmental disturbances as it was described in case of thalidomide. It may also cause immunomodulatory effects as it was shown for antibiotics, theobromine, caffeic acid or catechins on the pregnant mice model. At present, Echinacea purpurea-based phytoceuticals are among the most popular herbals in the marketplace. Many compounds of Echinacea extracts (polysaccharides, alkamides, polyphenols, glycoproteins exert immunomodulatory, anti-oxidative and anti-inflammatory activity. Echinacea is one of the most powerful and effective remedies against many kinds of bacterial and viral infections. In previous studies we shown significant inhibitory effect of the Echinacea purpurea based remedy on tumour angiogenic activity using cutaneous angiogenesis test, and an inhibitory effect on L-1 sarcoma growth was observed . The aim of the present study was to establish whether pharmaceuticals containing alcoholic extracts of Echinacea purpurea given to pregnant mice influence angiogenic activity and tissue VEGF and bFGF production of their fetuses. We showed that angiogenic activity of tissue homogenates was increased in Esberitox group and diminished in case of Immunal forte as compared to standard diet group. In case of Echinapur group we did not find significant differences in angiogenic activity. VEGF and bFGF concentration were lower in all groups compared to the control. In the case of Echinapur and Esberitox number of fetuses in one litter were slightly lower as compared to control group, but the difference is on the border of statistical significance. In conclusion, there is some possibility that pharmaceuticals containing Echinacea purpurea might influence fetal development in

  8. Caffeine administration alters the behaviour and development of Galleria mellonella larvae.

    Science.gov (United States)

    Maguire, Ronan; Kunc, Martin; Hyrsl, Pavel; Kavanagh, Kevin

    2017-11-01

    The effect of feeding caffeine on the behaviour and neural proteome of Galleria mellonella larvae was assessed. Caffeine was administered to larvae by force feeding and the metabolites theobromine and theophylline were subsequently detected by RP-HPLC analysis. Administration of caffeine to larvae resulted in reduced movement and a reduction in the formation of pupae. The production of the muscle relaxant theophylline may contribute to the reduction in larval movement. Analysis of the changes in proteome of the brain and surrounding tissues of caffeine fed larvae revealed an increase in the abundance of immune related proteins such as immune-related Hdd1 (6.28 fold increase) and hemolin (1.68 fold increase), ATPase associated proteins such as H+ transporting ATP synthase O subunit isoform 1 (1.87 fold increase) and H+ transporting ATP synthase delta subunit (1.53 fold increase) and proteins indicative of brain trauma such as troponin T transcript variant B, partial (1.55 fold increase). Proteins involved in development and protein degradation such as SUMO-activating enzyme subunit 1 (3.08 fold decrease) and chitin deacetylase, partial (3.67 fold decrease) were decreased in abundance. The results presented here indicate that caffeine is metabolised in a similar way in G. mellonella larvae to that in mammals and results in a variety of behavioural and developmental alterations. Utilisation of insects for studying the effects of caffeine and other neuroactive compounds may offer new insights into their mode of action and reduce the need to use mammals for this type of analysis. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Diversity of Cacao Trees in Waslala, Nicaragua: Associations between Genotype Spectra, Product Quality and Yield Potential

    Science.gov (United States)

    Trognitz, Bodo; Cros, Emile; Assemat, Sophie; Davrieux, Fabrice; Forestier-Chiron, Nelly; Ayestas, Eusebio; Kuant, Aldo; Scheldeman, Xavier; Hermann, Michael

    2013-01-01

    The sensory quality and the contents of quality-determining chemical compounds in unfermented and fermented cocoa from 100 cacao trees (individual genotypes) representing groups of nine genotype spectra (GG), grown at smallholder plantings in the municipality of Waslala, Nicaragua, were evaluated for two successive harvest periods. Cocoa samples were fermented using a technique mimicking recommended on-farm practices. The sensory cocoa quality was assessed by experienced tasters, and seven major chemical taste compounds were quantified by near infrared spectrometry (NIRS). The association of the nine, partially admixed, genotype spectra with the analytical and sensory quality parameters was tested. The individual parameters were analyzed as a function of the factors GG and harvest (including the date of fermentation), individual trees within a single GG were used as replications. In fermented cocoa, significant GG-specific differences were observed for methylxanthines, theobromine-to-caffeine (T/C) ratio, total fat, procyanidin B5 and epicatechin, as well as the sensory attributes global score, astringency, and dry fruit aroma, but differences related to harvest were also apparent. The potential cocoa yield was also highly determined by the individual GG, although there was significant tree-to-tree variation within every single GG. Non-fermented samples showed large harvest-to-harvest variation of their chemical composition, while differences between GG were insignificant. These results suggest that selection by the genetic background, represented here by groups of partially admixed genotype spectra, would be a useful strategy toward enhancing quality and yield of cocoa in Nicaragua. Selection by the GG within the local, genetically segregating populations of seed-propagated cacao, followed by clonal propagation of best-performing individuals of the selected GG could be a viable alternative to traditional propagation of cacao by seed from open pollination. Fast and

  10. Effect of chocolate and mate tea on the lipid profile of individuals with HIV/AIDS on antiretroviral therapy: A clinical trial.

    Science.gov (United States)

    Souza, Suelen J; Petrilli, Aline A; Teixeira, Andrea M; Pontilho, Patricia M; Carioca, Antonio A; Luzia, Liania A; Souza, José M; Damasceno, Nágila R; Segurado, Aluisio A; Rondó, Patricia H

    HIV/AIDS is generally associated with dyslipidemia and oxidative imbalance, which are caused by the infection itself and by antiretroviral therapy (ART). The flavonoids, found in cocoa and yerba mate, have antioxidant and hypolipidemic properties. The aim of this study was to evaluate the effects of the consumption of dark chocolate and mate tea on the lipid profiles of individuals with HIV/AIDS who are undergoing ART. A randomized, double-blind, placebo-controlled crossover clinical trial was conducted with 92 patients receiving ART for ≥6 mo and with viral suppression. The participants were randomized to receive either 65 g of chocolate (with 2148 mg polyphenols) or placebo chocolate (without polyphenols) or 3 g of mate tea (with 107 mg total phenols and 84.24 mg chlorogenic acid) or placebo mate (without polyphenols) for 15 d each, separated by a washout period of 15 d. The lipid profile, including determination of electronegative low-density lipoprotein, was determined after each intervention. The data were analyzed by analysis of variance using the pkcross procedure of the Stata 11.0 software. Analysis of variance revealed a significant overall difference in mean high-density lipoprotein cholesterol (HDL-C) between all supplements (P = 0.047). Using the paired t test, the effect was attributed to the consumption of dark chocolate (P = 0.046). The other parameters investigated were not improved. The consumption of dark chocolate for 15 d improved HDL-C concentrations of individuals with HIV/AIDS undergoing ART, possibly due to the presence of fatty acids (stearic acid), polyphenols, and theobromine. This fact is important for the cardiovascular protection of these individuals. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Methylxanthine Content in Commonly Consumed Foods in Spain and Determination of Its Intake during Consumption

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    Juan M. Sanchez

    2017-12-01

    Full Text Available Methylxanthines present psychostimulant effects. These compounds have low toxicity and their consumption at moderate levels presents some beneficial health effects, whereas some significant risk appears at high levels. Samples of common types of methylxanthine-containing beverages and foods consumed in Spain were analyzed to determine their content. Caffeine was the methylxanthine that was most found in the samples investigated. Instant coffees gave the highest caffeine percentage (18–44 mg·g−1. Green and scented teas were found to have a caffeine dry-weight content (8–26 mg·g−1 equivalent to ground coffees (13–23 mg·g−1, but black and pu-erh teas (18–30 mg·g−1 had a higher caffeine content. The evaluation of the most conventional methods for preparing espresso coffees showed that an espresso contains between 88–116 mg of caffeine. In the case of tea beverages, the amount of caffeine present was 2–3 times smaller than in espresso coffees. Energy drinks showed a similar caffeine content (80–106 mg as espresso coffees. Chocolates had the lowest caffeine content. It has been found that none of the foods evaluated reach the recommended daily intake limit of 400 mg of caffeine with a single dose. This limit can be reached with 4–5 doses in the case of coffees and energy drinks. In the case of chocolates, the methylxanthine compound detected at large levels was theobromine, with amounts ranging from 4 to 10 mg·g−1 for dark chocolates.

  12. Prehispanic Use of Chili Peppers in Chiapas, Mexico

    Science.gov (United States)

    Powis, Terry G.; Gallaga Murrieta, Emiliano; Lesure, Richard; Lopez Bravo, Roberto; Grivetti, Louis; Kucera, Heidi; Gaikwad, Nilesh W.

    2013-01-01

    The genus Capsicum is New World in origin and represents a complex of a wide variety of both wild and domesticated taxa. Peppers or fruits of Capsicum species rarely have been identified in the paleoethnobotanical record in either Meso- or South America. We report here confirmation of Capsicum sp. residues from pottery samples excavated at Chiapa de Corzo in southern Mexico dated from Middle to Late Preclassic periods (400 BCE to 300 CE). Residues from 13 different pottery types were collected and extracted using standard techniques. Presence of Capsicum was confirmed by ultra-performance liquid chromatography (UPLC)/MS-MS Analysis. Five pottery types exhibited chemical peaks for Capsicum when compared to the standard (dihydrocapsaicin). No peaks were observed in the remaining eight samples. Results of the chemical extractions provide conclusive evidence for Capsicum use at Chiapas de Corzo during a 700 year period (400 BCE–300 CE). Presence of Capsicum in different types of culinary-associated pottery raises questions how chili pepper could have been used during this early time period. As Pre-Columbian cacao products sometimes were flavored using Capsicum, the same pottery sample set was tested for evidence of cacao using a theobromine marker: these results were negative. As each vessel that tested positive for Capsicum had a culinary use we suggest here the possibility that chili residues from the Chiapas de Corzo pottery samples reflect either paste or beverage preparations for religious, festival, or every day culinary use. Alternatively, some vessels that tested positive merely could have been used to store peppers. Most interesting from an archaeological context was the presence of Capsicum residue obtained from a spouted jar, a pottery type previously thought only to be used for pouring liquids. PMID:24236083

  13. Time-course measurements of caffeine and its metabolites extracted from fingertips after coffee intake: a preliminary study for the detection of drugs from fingerprints.

    Science.gov (United States)

    Kuwayama, Kenji; Tsujikawa, Kenji; Miyaguchi, Hajime; Kanamori, Tatsuyuki; Iwata, Yuko T; Inoue, Hiroyuki

    2013-05-01

    The aim of this study was to determine whether an ingested drug and its metabolites could be detected in the subject's fingerprints. Caffeine (CF) was chosen as the model drug. Three healthy subjects were asked to consume a cup of coffee (ca. 100 mL) containing 80 micro micro mg CF as the total dose, which is the normal amount in one cup of coffee. After washing hands with water to remove external contaminants, each subject pressed the index fingertip to a collecting matrix just before consuming the test cup of coffee, and then again pressed the index fingertip to the collecting matrix after 1, 3, 5, and 7 h. The time curve of the amounts of CF and its metabolites-theobromine (TB), paraxanthine (PX), and theophylline (TP)-in fingerprints and blood was determined using liquid chromatography/tandem mass spectrometry (LC/MS). A filter paper wetted with water (50 μL) was an efficient collecting matrix for extracting the analytes from the fingertip. With optimized sample preparation and LC/MS conditions, the total operating time, from taking the fingerprints to obtaining the analytical result, was approximately 10 min. The lower limits of quantification for CF, TB, PX, and TP were 0.5, 5, 0.5, and 5 ng/fingerprint, respectively. The amount of CF or PX determined in fingerprints obtained over 7 h after coffee intake was significantly greater than the amount determined in fingerprints taken before drinking coffee. Fingerprints were a more efficient source for drug testing than other biological samples, such as blood and sweat, because the procedures for sampling and extracting the drugs were simpler and took less time. The method could be used to prove drug intake in criminal investigations.

  14. In vitro antioxidant properties, free radicals scavenging activities of extracts and polyphenol composition of a non-timber forest product used as spice: Monodora myristica

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    Bruno Moukette Moukette

    2015-01-01

    Full Text Available BACKGROUND: Excessive production of free radicals causes direct damage to biological molecules such as DNA, proteins, lipids, carbohydrates leading to tumor development and progression. Natural antioxidant molecules from phytochemicals of plant origin may directly inhibit either their production or limit their propagation or destroy them to protect the system. In the present study, Monodora myristica a non-timber forest product consumed in Cameroon as spice was screened for its free radical scavenging properties, antioxidant and enzymes protective activities. Its phenolic compound profile was also realized by HPLC. RESULTS: This study demonstrated that M. myristica has scavenging properties against DPPH',OH',NO', and ABTS'radicals which vary in a dose depending manner. It also showed an antioxidant potential that was comparable with that of Butylated Hydroxytoluene (BHT and vitamin C used as standard. The aqueous ethanol extract of M. myristica barks (AEH; showed a significantly higher content in polyphenolic compounds (21.44 ±0.24 mg caffeic acid/g dried extract and flavonoid (5.69 ± 0.07 quercetin equivalent mg/g of dried weight as compared to the other studied extracts. The HPLC analysis of the barks and leaves revealed the presence of several polyphenols. The acids (3,4-OH-benzoic, caffeic, gallic, O- and P- coumaric, syringic, vanillic, alcohols (tyrosol and OH-tyrosol, theobromine, quercetin, rutin, catechine and apigenin were the identified and quantified polyphenols. All the tested extracts demonstrated a high protective potential on the superoxide dismutase (SOD, catalase and peroxidase activities. CONCLUSION: Finally, the different extracts from M. myristica and specifically the aqueous ethanol extract reveal several properties such as higher free radical scavenging properties, significant antioxidant capacities and protective potential effects on liver enzymes.

  15. Interactions among chemical components of Cocoa tea (Camellia ptilophylla Chang), a naturally low caffeine-containing tea species.

    Science.gov (United States)

    Lin, Xiaorong; Chen, Zhongzheng; Zhang, Yuanyuan; Gao, Xiong; Luo, Wei; Li, Bin

    2014-06-01

    In the 1980s, a novel tea species, Cocoa tea (Camellia ptilophylla Chang), was discovered in Southern China with surprisingly low caffeine content (0.2% by dry weight). Although its health promoting characteristics have been known for a while, a very limited amount of scientific research has been focused on Cocoa tea. Herein, a systematic study on Cocoa tea and its chemical components, interactions and bioactivities was performed. YD tea (Yunnan Daye tea, Camellia sinensis), a tea species with a high caffeine content (5.8% by dry weight), was used as a control. By UV-Vis spectrometry, High Performance Liquid Chromatography (HPLC), and Flame Atomic Absorption Spectrometry (FAAS) for chemical composition analysis, C-2 epimeric isomers of tea catechins and theobromine were found to be the major catechins and methylxanthine in Cocoa tea, respectively. More gallated catechins, methylxanthines, and proteins were detected in Cocoa tea compared with YD tea. Moreover, the tendency of major components in Cocoa tea for precipitation was significantly higher than that in YD tea. Catechins, methylxanthines, proteins, iron, calcium, and copper were presumed to be the origins of molecular interactions in Cocoa tea and YD tea. The interactions between catechins and methylxanthines were highly related to the galloyl moiety in catechins and methyl groups in methylxanthines. In vitro anti-inflammatory activity assays revealed that Cocoa tea was a more potent inhibitor of nitric oxide (NO) in lipopolysaccharide (LPS)-stimulated macrophage cells (RAW 264.7) than YD tea. This study constructs a solid phytochemical foundation for further research on the mechanisms of molecular interactions and the integrated functions of Cocoa tea.

  16. Exhaustive Qualitative LC-DAD-MSn Analysis of Arabica Green Coffee Beans: Cinnamoyl-glycosides and Cinnamoylshikimic Acids as New Polyphenols in Green Coffee.

    Science.gov (United States)

    Baeza, Gema; Sarriá, Beatriz; Bravo, Laura; Mateos, Raquel

    2016-12-28

    Coffee is one of the most consumed beverages in the world, due to its unique aroma and stimulant properties. Although its health effects are controversial, moderate intake seems to be beneficial. The present work deals with the characterization and quantification of polyphenols and methylxanthines in four Arabica green coffee beans from different geographical origins. The antioxidant activity was also evaluated. Forty-three polyphenols (cinnamic acid, cinnamoyl-amide, 5 cinammoyl-glycosides, and 36 cinnamate esters) were identified using LC-MSn. Among these, cinnamate esters of six different chemical groups (including two dimethoxycinnamoylquinic acid isomers, three caffeoyl-feruloylquinic acid isomers, caffeoyl-sinapoylquinic acid, p-coumaroyl-feruloylquinic acid, two caffeoylshikimic acid isomers, and trimethoxycinnamoylshikimic acid) in addition to five isomers of cinnamoyl-glycosides called caffeoyl-2,7-anhydro-3-deoxy-2-octulopyranosic acid (CDOA) are described for the first time in Arabica green coffee beans. Moreover, 38 polyphenols (6-7% w/w) and 2 methylxanthines (1.3% w/w) were quantified by HPLC-DAD. Caffeoylquinic was the most abundant group of compounds (up to 85.5%) followed by dicaffeoylquinic and feruloylquinic acids (up to 8 and 7%, respectively) and the newly identified cinnamoyl-glycosides (CDOA) (up to 2.5%). Caffeine was the main methylxanthine (99.8%), with minimal amounts of theobromine (0.2%). African coffees (from Kenya and Ethiopia) showed higher polyphenolic content than American beans (from Brazil and Colombia), whereas methylxanthine contents varied randomly. Both phenols and methylxanthines contributed to the antioxidant capacity associated with green coffee, with a higher contribution of polyphenols. We conclude that green coffee represents an important source of polyphenols and methylxanthines, with high antioxidant capacity.

  17. Molecularly imprinted poly(4-amino-5-hydroxy-2,7-naphthalenedisulfonic acid) modified glassy carbon electrode as an electrochemical theophylline sensor

    Energy Technology Data Exchange (ETDEWEB)

    Aswini, K.K., E-mail: aswinikk@ymail.com; Vinu Mohan, A.M.; Biju, V.M., E-mail: vmbiju@ymail.com

    2016-08-01

    Theophylline is an inexpensive drug employed in asthma and chronic obstructive pulmonary disorder medications and is toxic at higher concentration. The development of a molecularly imprinted polymer based theophylline electrochemical sensor on glassy carbon electrode by the electropolymerization of 4-amino-5-hydroxy-2,7-naphthalenedisulfonic acid is being discussed in this work. The MIP modification enhances the theophylline recognition ability and the electron transfer kinetics of the bare electrode. The parameters, controlling the performance of the imprinted polymer based sensor, like number of electropolymerization cycles, composition of the pre-polymerization mixture, pH and immersion time were investigated and optimized. The interaction energy and the most stable conformation of the template–monomer complex in the pre-polymerization mixture were determined computationally using ab initio calculations based on density functional theory. The amperometric measurements showed that the developed sensor has a method detection limit of 0.32 μM for the dynamic range of 0.4 to 17 μM, at optimized conditions. The transducer possesses appreciable selectivity in the presence of structurally similar interferents such as theobromine, caffeine and doxofylline. The developed sensor showed remarkable stability and reproducibility and was also successfully employed in theophylline detection from commercially available tablets. - Highlights: • Molecularly imprinted polymer based theophylline sensor was developed. • Imprinted poly(4-amino-5-hydroxy-2,7-naphthalenedisulfonic acid) was electrodeposited. • Most stable template-monomer complex was assigned by computational analysis. • Possessed remarkable selectivity in the presence of structurally similar interferents • Employed for theophylline detection from commercially available tablets.

  18. Efecto del Tostado Sobre los Metabolitos Secundarios y la Actividad Antioxidante de Clones de Cacao Colombiano / Effect of Roasting on the Secondary Metabolites and Antioxidant Activity of Colombian Cocoa Clones

    Directory of Open Access Journals (Sweden)

    Sandra Zapata Bustamante

    2014-12-01

    Full Text Available Resumen. El tostado es una de las operaciones tecnológicas másimportantes en el procesamiento de los granos de cacao; esteproceso conduce a la formación de los productos de la reacciónde Maillard (RM los cuales son cruciales para el desarrollo dela calidad organoléptica de los granos de cacao y sus productosasociados. En el presente estudio se evaluó el efecto del tostadosobre el contenido de metabolitos secundarios y la actividadantioxidante en cinco clones de cacao cultivados en Colombia.En los extractos metanólicos de los diferentes clones de cacao sedeterminó el contenido de fenoles totales, taninos condensados,antocianinas totales, catequina y epicatequina, teobromina ycafeína. Además, se determinó la actividad antioxidante pormedio de la actividad atrapadora del radical ABTS•+, el métodoORAC y la capacidad atrapadora de radicales superóxido. Enconclusión, el efecto del tostado en los clones de cacao no tuvo uncomportamiento uniforme sobre los cambios en los contenidos delos diversos metabolitos secundarios y la actividad antioxidante. /  Abstract. Roasting is one of the most important technologicaloperations in the processing of cocoa beans; this process leads tothe formation of products of the Maillard reaction (RM, which arecrucial for the development of organoleptic quality of cocoa beansand its associated products. The aim of this study was to evaluatethe effect of roasting on the content of secondary metabolites andantioxidant activity in five Colombian cocoa clones, by differentmethodologies. The methanolic extracts of the cocoa beans wereanalyzed for total phenols content, condensed tannins, totalantiocianins, catechin and epicatechin, theobromine and caffeine.The antioxidant activity was determined by ABTS assay, ORACmethod and superoxide radical scavenging method. The effect ofroasting on cocoa clones did not have an unspecified behavior. Inconclusion, the effect of roasting of cacao clones was not uniformon the

  19. Effects of phenothiazine neuroleptics on the rate of caffeine demethylation and hydroxylation in the rat liver.

    Science.gov (United States)

    Daniel, W A; Syrek, M; Ryłko, Z; Kot, M

    2001-01-01

    The primary metabolic pathways of caffeine are 3-N-demethylation to paraxanthine (CYP1A2), 1-N-demethylation to theobromine and 7-N-demethylation to theophylline (CYP1A2 and other enzymes), and 8-hydroxylation to 1,3,7-trimethyluric acid (CYP3A). The aim of the present study was to investigate the influence of phenothiazine neuroleptics (chlorpromazine, levomepromazine, thioridazine, perazine) on cytochrome P-450 activity measured by caffeine oxidation in rat liver microsomes. The obtained results showed that all the investigated neuroleptics competitively inhibited caffeine oxidation in the rat liver, though their potency to inhibit particular metabolic pathways was not equal. Levomepromazine exerted the most potent inhibitory effect on caffeine oxidation pathways, the effect on 8-hydroxylation being the most pronounced. This indicates inhibition of CYP 1 A2 (inhibition of 3-N- and 1-N-demethylation; Ki = 36 and 32 microM, respectively), CYP3A2 (inhibition of 8-hydroxylations; Ki = 20 microM), and possibly other CYP isoenzymes (inhibition of 7-N-demethylation; Ki = 58 microM) by the neuroleptics. The potency of inhibition of caffeine oxidation by perazine was similar to levomepromazine. Thioridazine was a weaker inhibitor of caffeine 3-N- and 7-N-demethylation, while chlorpromazine was weaker in inhibiting caffeine 1-N- and 7-N-demethylation, compared to levomepromazine. In summary, the obtained results showed that all the investigated neuroleptics had a broad spectra of CYP inhibition in the rat liver. The isoenzymes CYP1A2 and CYP3A2 were distinctly inhibited by all the investigated neuroleptics, while other CYP isoenzymes (CYP2B and/or 2E1) by perazine and levomepromazine. The CYP3A2 inhibition was most pronounced. (Ki = 20-40 microM).

  20. Investigation of free amino acid, total phenolics, antioxidant activity and purine alkaloids to assess the health properties of non-Camellia tea

    Directory of Open Access Journals (Sweden)

    Wu Bi

    2016-03-01

    Full Text Available To find novel functional beverages from folk teas, 33 species of frequently used non-Camellia tea (plants other than Camellia were collected and compared with Camellia tea (green tea, pu-erh tea and black tea for the first time. Data are reported here on the quantities of 20 free amino acids (FAAs and three purine alkaloids (measured by UHPLC, total polyphenols (measured by Folin-Ciocalteu assay, and antioxidant activity (DPPH. The total amounts of FAAs in non-Camellia tea (0.62–18.99 mg/g are generally less than that of Camellia tea (16.55–24.99 mg/g. However, for certain FAAs, the quantities were much higher in some non-Camellia teas, such as γ-aminobutyric acid in teas from Ampelopsis grossedentata, Isodon serra and Hibiscus sabdariffa. Interestingly, theanine was detected in tea from Potentilla fruticosa (1.16±0.81 mg/g. Furthermore, the content of polyphenols in teas from A. grossedentata, Acer tataricum subsp. ginnala are significantly higher than those from Camellia tea; teas from I. serra, Pistacia chinensis and A. tataricum subsp. ginnala have remarkable antioxidant activities similar to the activities from green tea (44.23 μg/mL. Purine alkaloids (caffeine, theobromine and theophylline were not detected in non-Camellia teas. The investigation suggest some non-Camellia teas may be great functional natural products with potential for prevention of chronic diseases and aging, by providing with abundant polyphenols, antioxidants and specific FAAs.

  1. Diversity of cacao trees in Waslala, Nicaragua: associations between genotype spectra, product quality and yield potential.

    Directory of Open Access Journals (Sweden)

    Bodo Trognitz

    Full Text Available The sensory quality and the contents of quality-determining chemical compounds in unfermented and fermented cocoa from 100 cacao trees (individual genotypes representing groups of nine genotype spectra (GG, grown at smallholder plantings in the municipality of Waslala, Nicaragua, were evaluated for two successive harvest periods. Cocoa samples were fermented using a technique mimicking recommended on-farm practices. The sensory cocoa quality was assessed by experienced tasters, and seven major chemical taste compounds were quantified by near infrared spectrometry (NIRS. The association of the nine, partially admixed, genotype spectra with the analytical and sensory quality parameters was tested. The individual parameters were analyzed as a function of the factors GG and harvest (including the date of fermentation, individual trees within a single GG were used as replications. In fermented cocoa, significant GG-specific differences were observed for methylxanthines, theobromine-to-caffeine (T/C ratio, total fat, procyanidin B5 and epicatechin, as well as the sensory attributes global score, astringency, and dry fruit aroma, but differences related to harvest were also apparent. The potential cocoa yield was also highly determined by the individual GG, although there was significant tree-to-tree variation within every single GG. Non-fermented samples showed large harvest-to-harvest variation of their chemical composition, while differences between GG were insignificant. These results suggest that selection by the genetic background, represented here by groups of partially admixed genotype spectra, would be a useful strategy toward enhancing quality and yield of cocoa in Nicaragua. Selection by the GG within the local, genetically segregating populations of seed-propagated cacao, followed by clonal propagation of best-performing individuals of the selected GG could be a viable alternative to traditional propagation of cacao by seed from open

  2. Facilitating high resolution mass spectrometry data processing for screening of environmental water samples: An evaluation of two deconvolution tools

    Energy Technology Data Exchange (ETDEWEB)

    Bade, Richard [Research Institute for Pesticides and Water, University Jaume I, Avda. Sos Baynat s/n, E-12071 Castellón (Spain); Causanilles, Ana; Emke, Erik [KWR Watercycle Research Institute, Chemical Water Quality and Health, P.O. Box 1072, 3430 BB Nieuwegein (Netherlands); Bijlsma, Lubertus; Sancho, Juan V.; Hernandez, Felix [Research Institute for Pesticides and Water, University Jaume I, Avda. Sos Baynat s/n, E-12071 Castellón (Spain); Voogt, Pim de, E-mail: w.p.devoogt@uva.nl [KWR Watercycle Research Institute, Chemical Water Quality and Health, P.O. Box 1072, 3430 BB Nieuwegein (Netherlands); Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam, P.O. Box 94248, 1090 GE Amsterdam (Netherlands)

    2016-11-01

    A screening approach was applied to influent and effluent wastewater samples. After injection in a LC-LTQ-Orbitrap, data analysis was performed using two deconvolution tools, MsXelerator (modules MPeaks and MS Compare) and Sieve 2.1. The outputs were searched incorporating an in-house database of > 200 pharmaceuticals and illicit drugs or ChemSpider. This hidden target screening approach led to the detection of numerous compounds including the illicit drug cocaine and its metabolite benzoylecgonine and the pharmaceuticals carbamazepine, gemfibrozil and losartan. The compounds found using both approaches were combined, and isotopic pattern and retention time prediction were used to filter out false positives. The remaining potential positives were reanalysed in MS/MS mode and their product ions were compared with literature and/or mass spectral libraries. The inclusion of the chemical database ChemSpider led to the tentative identification of several metabolites, including paraxanthine, theobromine, theophylline and carboxylosartan, as well as the pharmaceutical phenazone. The first three of these compounds are isomers and they were subsequently distinguished based on their product ions and predicted retention times. This work has shown that the use deconvolution tools facilitates non-target screening and enables the identification of a higher number of compounds. - Highlights: • A hidden target non-target screening method is utilised using two databases • Two software (MsXelerator and Sieve 2.1) used for both methods • 22 compounds tentatively identified following MS/MS reinjection • More information gleaned from this combined approach than individually.

  3. Look what I found! Poison hunting on eBay.

    Science.gov (United States)

    Cantrell, F Lee

    2005-01-01

    Many substances deemed too dangerous for commercial use are still available to the general public. The purchase of these substances may potentially place members of the general public at risk for serious poisonings. This study was designed to document the large variety of dangerous poisons readily available on a popular online auction Web site. Methods. Over a 10-month period, the online auction Web site eBays was searched daily using the terms "poison" and "contents." Product name, active ingredients, what form the product is in, amount in container, and relative toxicity rating (Clinical Toxicology of Commercial Products, Gosselin, et al.) were recorded. If available, pictures of the products were saved. One hundred twenty-one individual products were identified. Fifty-five were in solid/tablet form, 37 were powders, and 29 were liquids. Product containers were full for 56 items and partially full for 65. Twenty-four products contained ingredients rated as "supertoxic" and included strychnine (10), arsenic trioxide (8), cyanide (2) and nicotine, pilocarpine, phosphorus, powdered conium maculatum (1 each). Sixty-three products contained "extremely toxic" ingredients including thallium, picrotoxin, soluble barium, antimony, mercury, arsenates, podophyllin, fluoride, zinc phosphide, atropine, scopolamine, and plant extracts of gelsemium, aconite, larkspur, and croton. Twenty-one products contained "very toxic" ingredients including lead, copper, camphor, caffeine, theobromine, creosote, pyrogallic acid, sparteine, quinine, lindane, warfarin, phenol, and digitalis. The remaining 13 were "moderately-slightly toxic." While the viability of the labeled ingredients could not be verified, the transportation, handling, and potential utilization of these dangerous poisons by the general public could result in serious poisonings.

  4. Effect of age and smoking on in vivo CYP1A2, flavin-containing monooxygenase, and xanthine oxidase activities in Koreans: determination by caffeine metabolism.

    Science.gov (United States)

    Chung, W G; Kang, J H; Park, C S; Cho, M H; Cha, Y N

    2000-03-01

    To assess the effect of gender, age, and smoking habits on the in vivo activities of CYP1A2, flavin-containing monooxygenase (FMO), and xanthine oxidase in Korean subjects. One hundred thirty-three age- and gender-matched healthy Korean volunteers (age range, 21 to 78 years; mean age, 35.3 +/- 16.6 years) with and without smoking habits participated. After drinking a cup of coffee (200 mL) that contained 110 mg caffeine, a 1-hour urine sample (between 4 and 5 hours) was collected and caffeine metabolites were analyzed by HPLC. There were marked individual variations in CYP1A2 [(1,7-dimethylurate + paraxanthine)/caffeine], FMO (theobromine/caffeine), and xanthine oxidase (1-methylurate/1-methylxanthine) activities (14-, 42-, and 9-fold, respectively). However, the mean values of these enzyme activities in the nonsmokers were not different between men and women. In the nonsmoking subjects in their 20s, the mean values of CYP1A2 and FMO activities (13.5 +/- 5.9 and 2.1 +/- 1.9, respectively) were higher than those (7.9 +/-1.8 and 0.95 +/- 0.22) of older decennial age groups. Xanthine oxidase activities were the same for all age groups (subjects in their 20s through their 70s). CYP1A2 activity of the smokers (20.0 +/- 9.6) was higher than that of the nonsmokers (10.8 +/- 5.8; P Korean volunteers with and without smoking habits provided the baseline and the widely varying interindividual activities of CYP1A2, FMO, and xanthine oxidase in a Korean population. The results also suggested that drugs metabolized by CYP1A2 and FMO may require individualized dose adjustment according to the age and smoking habits of the subjects.

  5. Methylxanthine Content in Commonly Consumed Foods in Spain and Determination of Its Intake during Consumption.

    Science.gov (United States)

    Sanchez, Juan M

    2017-12-04

    Methylxanthines present psychostimulant effects. These compounds have low toxicity and their consumption at moderate levels presents some beneficial health effects, whereas some significant risk appears at high levels. Samples of common types of methylxanthine-containing beverages and foods consumed in Spain were analyzed to determine their content. Caffeine was the methylxanthine that was most found in the samples investigated. Instant coffees gave the highest caffeine percentage (18-44 mg·g-1). Green and scented teas were found to have a caffeine dry-weight content (8-26 mg·g-1) equivalent to ground coffees (13-23 mg·g-1), but black and pu-erh teas (18-30 mg·g-1) had a higher caffeine content. The evaluation of the most conventional methods for preparing espresso coffees showed that an espresso contains between 88-116 mg of caffeine. In the case of tea beverages, the amount of caffeine present was 2-3 times smaller than in espresso coffees. Energy drinks showed a similar caffeine content (80-106 mg) as espresso coffees. Chocolates had the lowest caffeine content. It has been found that none of the foods evaluated reach the recommended daily intake limit of 400 mg of caffeine with a single dose. This limit can be reached with 4-5 doses in the case of coffees and energy drinks. In the case of chocolates, the methylxanthine compound detected at large levels was theobromine, with amounts ranging from 4 to 10 mg·g-1 for dark chocolates.

  6. Determination of theophylline by HPLC and GC-IDMS, the effect of chemically similar xanthine derivatives on the specificity of the method and the possibility of paracetamol as interfering substance.

    Science.gov (United States)

    Kress, Michael; Meissner, Dieane; Kaiser, Patricia; Hanke, Rainer; Wood, William Graham

    2002-01-01

    The aim of this study was to develop and compare high-performance liquid chromatography (HPLC) and gas chromatography coupled with isotope dilution-mass spectrometry (GC-IDMS) methods with a common extraction procedure for the determination of substituted xanthines in biological matrices such as serum and urine. For HPLC both isocratic and gradient methods were evaluated. Difficulties occurred in separation of all 6 xanthines of interest - uric acid, theobromine, theophylline, paraxanthine, caffeine and 1,3-dimethyl-7-(2-hydroxyethyl) xanthine as internal standard. In addition, paracetamol was seen to interfere at higher concentrations, which meant that a system had to be developed to separate all 7 components of interest. The final solution chosen consisted of precipitation of serum samples with 6 mol/l trichloroacetic acid followed by neutralisation with 3 mol/l KOH and chromatography on a 150 x 4.6 mm Nautilus C-18 column (Macherey & Nagel) using an isocratic elution consisting of 0.02 mol/l acetate-phosphate buffer, pH 3.0 containing 9.6% v/v acetonitrile and monitoring at 273 +/- 7 nm. Comparisons with GC-IDMS and FPIA were acceptable. Run times of 10 minutes were possible. An additional "safe time" of 5 minutes was allowed to elute any substances with similar absorption maxima which were sometimes present in commercial control sera. Precision of the method was 1.64% (intra-assay) and 2.87% (inter-assay) at 4.1 mg/l and 1.51% respectively 2.15% at 25 mg/l including extraction and measurement steps. Recovery was between 86 and 101% between 1.25 and 100 mg/l and peak time deviations for all 7 components between 0.07% and 0.34% (coefficient of variation) in 7 consecutive measurements.

  7. Acefylline activates filaggrin deimination by peptidylarginine deiminases in the upper epidermis.

    Science.gov (United States)

    Méchin, Marie-Claire; Cau, Laura; Galliano, Marie-Florence; Daunes-Marion, Sylvie; Poigny, Stéphane; Vidaluc, Jean-Louis; Bessou-Touya, Sandrine; Takahara, Hidenari; Serre, Guy; Duplan, Hélène; Simon, Michel

    2016-02-01

    Peptidylarginine deiminases (PADs) catalyze deimination (or citrullination), a calcium-dependent post-translational modification involved in several physiological processes and human diseases, such as rheumatoid arthritis and cancer. Deimination of filaggrin (FLG) by PAD1 and PAD3 during the last steps of keratinocyte differentiation is a crucial event for the epidermis function and homeostasis. This allows the complete degradation of FLG, leading to the production of free amino acids and their derivatives that are essential for epidermal photoprotection and moisturizing of the stratum corneum. To increase the flux of this catabolic pathway, we searched for activators of PADs. A large chemical library was screened first in silico and then by using an automated assay based on an indirect colorimetric measurement of recombinant human PAD activity. Potential activators were then confirmed using a recombinant human FLG as a substrate, and secondly after topical application at the surface of three-dimensional reconstructed human epidermis. The data obtained after the library screening pointed to xanthine derivatives as potential PAD activators. Among seven xanthine derivatives tested at 50-300μM, caffeine, theobromine and acefylline proved to be the most potent enhancers of in vitro deimination of FLG by PAD1 and PAD3. After topical application of a gel formulation containing 3% acefylline at the surface of reconstructed epidermis, immunoblotting analysis showed an increase in the total amount of deiminated proteins, and confocal microscopy showed an enhanced deimination in the stratum corneum. This demonstrated the activation of PADs in living cells. As a PAD activator, acefylline will be useful to study the role of deimination and could be proposed to increase or correct the hydration of the cornified layers of the epidermis. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  8. Cocoa Procyanidins Suppress Transformation by Inhibiting Mitogen-activated Protein Kinase Kinase*S⃞

    Science.gov (United States)

    Kang, Nam Joo; Lee, Ki Won; Lee, Dong Eun; Rogozin, Evgeny A.; Bode, Ann M.; Lee, Hyong Joo; Dong, Zigang

    2008-01-01

    Cocoa was shown to inhibit chemically induced carcinogenesis in animals and exert antioxidant activity in humans. However, the molecular mechanisms of the chemopreventive potential of cocoa and its active ingredient(s) remain unknown. Here we report that cocoa procyanidins inhibit neoplastic cell transformation by suppressing the kinase activity of mitogen-activated protein kinase kinase (MEK). A cocoa procyanidin fraction (CPF) and procyanidin B2 at 5 μg/ml and 40 μm, respectively, inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced neoplastic transformation of JB6 P+ mouse epidermal (JB6 P+) cells by 47 and 93%, respectively. The TPA-induced promoter activity and expression of cyclooxygenase-2, which is involved in tumor promotion and inflammation, were dose-dependently inhibited by CPF or procyanidin B2. The activation of activator protein-1 and nuclear factor-κB induced by TPA was also attenuated by CPF or procyanidin B2. The TPA-induced phosphorylation of MEK, extracellular signal-regulated kinase, and p90 ribosomal s6 kinase was suppressed by CPF or procyanidin B2. In vitro and ex vivo kinase assay data demonstrated that CPF or procyanidin B2 inhibited the kinase activity of MEK1 and directly bound with MEK1. CPF or procyanidin B2 suppressed JB6 P+ cell transformation induced by epidermal growth factor or H-Ras, both of which are known to be involved in MEK/ERK signal activation. In contrast, theobromine (up to 80 μm) had no effect on TPA-induced transformation, cyclooxygenase-2 expression, the transactivation of activator protein-1 or nuclear factor-κB, or MEK. Notably, procyanidin B2 exerted stronger inhibitory effects compared with PD098059 (a well known pharmacological inhibitor of MEK) on MEK1 activity and neoplastic cell transformation. PMID:18519570

  9. Dietary nutrients associated with short and long sleep duration. Data from a nationally representative sample☆

    Science.gov (United States)

    Grandner, Michael A.; Jackson, Nicholas; Gerstner, Jason R.; Knutson, Kristen L.

    2013-01-01

    Short sleep duration is associated with weight gain and obesity, diabetes, cardiovascular disease, psychiatric illness, and performance deficits. Likewise, long sleep duration is also associated with poor physical and mental health. The role of a healthy diet in habitual sleep duration represents a largely unexplored pathway linking sleep and health. This study evaluated associations between habitual sleep parameters and dietary/nutritional variables obtained via the National Health and Nutrition Examination Survey (NHANES), 2007–2008. We hypothesized that habitual very short (theobromine (long sleep RR = 0.910, p < 0.05), vitamin C (short sleep RR = 0.890, p < 0.05), tap water (short sleep RR = 0.952, p < 0.001; very short (<5 h) sleep RR = 0.941, p < 0.05), lutein + zeaxanthin (short sleep RR = 1.123, p < 0.05), dodecanoic acid (long sleep RR = 0.812, p < 0.05), choline (long sleep RR = 0.450, p = 0.001), lycopene (very short (<5 h) sleep RR = 0.950, p <0.05), total carbohydrate (very short (<5 h) sleep RR = 0.494, p <0.05; long sleep RR = 0.509, p <0.05), selenium (short sleep RR = 0.670, p <0.01) and alcohol (long sleep RR = 1.172, p < 0.01). Overall, many nutrient variables were associated with short and/or long sleep duration, which may be explained by differences in food variety. Future studies should assess whether these associations are due to appetite dysregulation, due to short/long sleep and/or whether these nutrients have physiologic effects on sleep regulation. In addition, these data may help us better understand the complex relationship between diet and sleep and the potential role of diet in the relationship between sleep and obesity and other cardiometabolic risks. PMID:23339991

  10. Prehispanic use of chili peppers in Chiapas, Mexico.

    Directory of Open Access Journals (Sweden)

    Terry G Powis

    Full Text Available The genus Capsicum is New World in origin and represents a complex of a wide variety of both wild and domesticated taxa. Peppers or fruits of Capsicum species rarely have been identified in the paleoethnobotanical record in either Meso- or South America. We report here confirmation of Capsicum sp. residues from pottery samples excavated at Chiapa de Corzo in southern Mexico dated from Middle to Late Preclassic periods (400 BCE to 300 CE. Residues from 13 different pottery types were collected and extracted using standard techniques. Presence of Capsicum was confirmed by ultra-performance liquid chromatography (UPLC/MS-MS Analysis. Five pottery types exhibited chemical peaks for Capsicum when compared to the standard (dihydrocapsaicin. No peaks were observed in the remaining eight samples. Results of the chemical extractions provide conclusive evidence for Capsicum use at Chiapas de Corzo during a 700 year period (400 BCE-300 CE. Presence of Capsicum in different types of culinary-associated pottery raises questions how chili pepper could have been used during this early time period. As Pre-Columbian cacao products sometimes were flavored using Capsicum, the same pottery sample set was tested for evidence of cacao using a theobromine marker: these results were negative. As each vessel that tested positive for Capsicum had a culinary use we suggest here the possibility that chili residues from the Chiapas de Corzo pottery samples reflect either paste or beverage preparations for religious, festival, or every day culinary use. Alternatively, some vessels that tested positive merely could have been used to store peppers. Most interesting from an archaeological context was the presence of Capsicum residue obtained from a spouted jar, a pottery type previously thought only to be used for pouring liquids.

  11. Spectroscopic evidence of xanthine compounds fluorescence quenching effect on water-soluble porphyrins

    Science.gov (United States)

    Makarska-Bialokoz, Magdalena

    2015-02-01

    The formation of π-stacked complexes between water-soluble porphyrins: 4,4‧,4″,4″‧-(21H,23H-porphine-5,10,15,20-tetrayl)tetrakis-(benzoic acid) (H2TCPP), 5,10,15,20-tetrakis(4-sulfonatophenyl)-21H,23H-porphine (H2TPPS4), 5,10,15,20-tetrakis[4-(trimethylammonio)phenyl]-21H,23H-porphine tetra-p-tosylate (H2TTMePP), 5,10,15,20-tetrakis(1-methyl-4-pyridyl)-21H,23H-porphine tetra-p-tosylate (H2TMePyP), the Cu(II) complexes of H2TTMePP and H2TMePyP, as well as chlorophyll a with xanthine, theophylline (1,3-dimethylxanthine) and theobromine (3,7-dimethylxanthine) has been studied analysing their absorption and steady-state fluorescence spectra in aqueous (or acetone in case of chlorophyll a) solution. During titration by the compounds from xanthine group the bathochromic effect in the porphyrin absorption spectra as well as the hypochromicity of the porphyrin Soret maximum can be noticed. The fluorescence quenching effect observed during interactions in the systems examined suggests the process of static quenching. The association and fluorescence quenching constants are of the order of magnitude of 103 - 102 mol-1. The results obtained show that xanthine and its derivatives can quench the fluorescence of the porphyrins according to the number of methyl groups in the molecule of quencher.

  12. Dataset on exogenous application of salicylic acid and methyljasmonate and the accumulation of caffeine in young leaf tissues and catabolically inactive endosperms

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    Avinash Kumar

    2017-08-01

    Full Text Available Exogenous exposure of coffee plants to 50 μM and 500 μM salicylic acid through liquid hydroponic medium or the exposure to volatile fumes of methyljasmonate was carried out to study the role of salicylic acid and methyljasmonate on the accumulation of caffeine and other methylxanthines like 7-methylxanthine, theobromine and theophylline. Transcript levels of the first, second and third N-methyltransferase involved in the core caffeine biosynthetic pathway namely, xanthosine methyltransferase (XMT, methylxanthine methyltransferase (MXMT and di-methylxanthine methyltransferase (DXMT was investigated by semi-quantitative RT-PCR for validating the reason behind the changes of caffeine biosynthetic potential under the influence of the two analogues of plant phytohormones. Maturing coffee fruits are known to be biologically inactive with respect to caffeine biosynthetic activity in the endosperms. To understand this, fruits were treated with different doses of salicylic acid in a time-course manner and the de-repression of tissue maturation-mediated knockdown of caffeine biosynthesis by exogenously applied salicylic acid was achieved. In our companion paper [1] it was shown that the repression of NMT genes during the dry weight accumulation phase of maturing endosperm could be relaxed by the exogenous application of salicylic acid and methyljasmonate. A probable model based on the work carried out therein and based on other literature [2–4] was proposed to describe that the crosstalk between salicylic acid or methyljasmonate and the ABA/ethylene pathway and might involve transcription factors downstream to the signaling cascade.

  13. Correlation Analysis of Cocoa Consumption Data with Worldwide Incidence Rates of Testicular Cancer and Hypospadias

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    Fabrizio Giannandrea

    2009-02-01

    Full Text Available The underlying reasons for the increasing occurrence of male reproductive diseases (MRD such as hypospadias, cryptorchidism, and testicular cancer (TC over the last decades are still unknown. It has been hypothesized that the risk of MRD is determined in utero and that pregnancy dietary intake could also affect MRD risk in the offspring. Various studies in animals reported that cocoa and theobromine, the main stimulant of cocoa, exert toxic effects on the testis, inducing testicular atrophy and impaired sperm quality. A correlation analysis was conducted to examine the possible role of cocoa consumption on the occurrence of selected MRD during the prenatal and early life period of cases. The incidence rates between 1998-2002 of TC in 18 countries obtained from Cancer Incidence in Five Continents were correlated with the average per-capita consumption of cocoa (kg/capita/year (FAOSTAT-Database in these countries from 1965 to 1980, i.e. the period corresponding to the early life of TC cases. In order to test the above correlation in the case of hypospadias, the mean prevalence at birth in 20 countries (1999-2003 with average per-capita consumption of cocoa in these countries in the same period corresponding to pregnancy were used. The consumption of cocoa in the period 1965–80, was most closely correlated with the incidence of TC in young adults (r=0.859; p<0.001. An analogous significant correlation was also observed between early cocoa consumption and the prevalence rates of hypospadias in the period 1999-2003 (r=0.760; p<0.001. Although the ecological approach used in this study cannot provide an answer on the causal relationship between consumption of cocoa in early life and TC and hypospadias, the results are suggestive and indicate the need of further analytic studies to investigate the role of individual exposure to cocoa, particularly during the prenatal and in early life of the patients.

  14. A Preliminary Urinary Metabolomics Study of Sprague-Dawley Rats after Short-term Ketamine Administration by Proton Nuclear Magnetic Resonance Spectroscopy

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    Xiang Lu

    2016-01-01

    Full Text Available Drug abuse has become a global problem. The mass spectrometry-based metabolic consequences of ketamine administration in anesthesia and therapy have been well studied, but to the best of our knowledge, metabolomic studies of ketamine abuse based on nuclear magnetic resonance (NMR spectroscopy are still lacking. In this study, twenty Sprague–Dawley rats were randomly assigned into two groups: a control group (n = 10 and a ketamine group (n = 10. The animals in the ketamine group received intraperitoneal injections of ketamine twice daily at 12-h intervals at progressively increasing doses over a period of 9 days, while the control group received an equal volume of saline. The urine samples were collected for 24 h at days 0, 1, 3, 5, 7, and 9 for the metabolomics study. The metabolic changes in urine after short-term ketamine administration were analyzed by proton NMR coupled with multivariate statistical analysis. The results indicated that short-term ketamine exposure led to significant alterations of the metabolites in the urine of the rats. Specifically, 1,3,7-trimethyluric acid, 1,3-dimethyluric acid, acetoacetic acid, acetylglycine, creatine, sarcosine, dimethylglycine, glycine, and theobromine were significantly increased in the urine. Significant changes were also found in metabolites related to antioxidant and energy metabolism, including acetoacetic acid, succinate, 1,3,7-trimethyluric acid, 1,3-dimethyluric acid, creatine, and taurine. Our findings indicated that short-term ketamine administration leads to disorder of energy metabolism and oxidative stress. In addition, the modified metabolites identified could serve as the new biological markers and potential biological indices reflecting the underlying mechanism of ketamine abuse.

  15. Development of a biosensor for caffeine.

    Science.gov (United States)

    Babu, V R Sarath; Patra, S; Karanth, N G; Kumar, M A; Thakur, M S

    2007-01-23

    We have utilized a microbe, which can degrade caffeine to develop an Amperometric biosensor for determination of caffeine in solutions. Whole cells of Pseudomonas alcaligenes MTCC 5264 having the capability to degrade caffeine were immobilized on a cellophane membrane with a molecular weight cut off (MWCO) of 3000-6000 by covalent crosslinking method using glutaraledhyde as the bifunctional crosslinking agent and gelatin as the protein based stabilizing agent (PBSA). The biosensor system was able to detect caffeine in solution over a concentration range of 0.1 to 1 mg mL(-1). With read-times as short as 3 min, this caffeine biosensor acts as a rapid analysis system for caffeine in solutions. Interestingly, successful isolation and immobilization of caffeine degrading bacteria for the analysis of caffeine described here was enabled by a novel selection strategy that incorporated isolation of caffeine degrading bacteria capable of utilizing caffeine as the sole source of carbon and nitrogen from soils and induction of caffeine degrading capacity in bacteria for the development of the biosensor. This biosensor is highly specific for caffeine and response to interfering compounds such as theophylline, theobromine, paraxanthine, other methyl xanthines and sugars was found to be negligible. Although a few biosensing methods for caffeine are reported, they have limitations in application for commercial samples. The development and application of new caffeine detection methods remains an active area of investigation, particularly in food and clinical chemistry. The optimum pH and temperature of measurement were 6.8 and 30+/-2 degrees C, respectively. Interference in analysis of caffeine due to different substrates was observed but was not considerable. Caffeine content of commercial samples of instant tea and coffee was analyzed by the biosensor and the results compared well with HPLC analysis.

  16. Effects of paraxanthine and caffeine on sleep, locomotor activity, and body temperature in orexin/ataxin-3 transgenic narcoleptic mice.

    Science.gov (United States)

    Okuro, Masashi; Fujiki, Nobuhiro; Kotorii, Nozomu; Ishimaru, Yuji; Sokoloff, Pierre; Nishino, Seiji

    2010-07-01

    Caffeine, an adenosine A1 and A2a receptor antagonist, is a widely consumed stimulant and also used for the treatment of hypersomnia; however, the wake-promoting potency of caffeine is often not strong enough, and high doses may induce side effects. Caffeine is metabolized to paraxanthine, theobromine, and theophylline. Paraxanthine is a central nervous stimulant and exhibits higher potency at A1 and A2 receptors, but has lower toxicity and lesser anxiogenic effects than caffeine. We evaluated the wake-promoting efficacy of paraxanthine, caffeine, and a reference wake-promoting compound, modafinil, in a mice model of narcolepsy, a prototypical disease model of hypersomnia. Orexin/ataxin-3 transgenic (TG) and wild-type (WT) mice were subjected to oral administration (at ZT 2 and ZT14) of 3 doses of paraxanthine, caffeine, modafinil, or vehicle. Paraxanthine, caffeine, and modafinil significantly promoted wakefulness in both WT and narcoleptic TG mice and proportionally reduced NREM and REM sleep in both genotypes. The wake-promoting potency of 100 mg/kg p.o. of paraxanthine during the light period administration roughly corresponds to that of 200 mg/kg p.o. of modafinil. The wake-promoting potency of paraxanthine is greater and longer lasting than that of the equimolar concentration of caffeine, when the drugs were administered during the light period. The wake-promotion by paraxanthine, caffeine, and modafinil are associated with an increase in locomotor activity and body temperature. However, the higher doses of caffeine and modafinil, but not paraxanthine, induced hypothermia and reduced locomotor activity, thereby confirming the lower toxicity of paraxanthine. Behavioral evaluations of anxiety levels in WT mice revealed that paraxanthine induced less anxiety than caffeine did. Because it is also reported to provide neuroprotection, paraxanthine may be a better wake-promoting agent for hypersomnia associated with neurodegenerative diseases.

  17. Natural allelic variations of TCS1 play a crucial role in caffeine biosynthesis of tea plant and its related species.

    Science.gov (United States)

    Jin, Ji-Qiang; Yao, Ming-Zhe; Ma, Chun-Lei; Ma, Jian-Qiang; Chen, Liang

    2016-03-01

    Tea caffeine synthase 1 (TCS1) is an enzyme that catalyzes the methylation of N-3 and N-1 and considered to be the most critical enzyme in the caffeine biosynthetic pathway of tea plant. This study shows that TCS1 has six types of allelic variations, namely, TCS1a, TCS1b, TCS1c, TCS1d, TCS1e, and TCS1f, with a 252 bp insertion/deletion mutation in the 5'-untranslated region. Among tea plant and its related species, TCS1a is the predominant allele, and TCS1b-f are the rare alleles that mainly appear in few wild germplasms. The full-length cDNA sequences of three new alleles, namely, TCS1d, TCS1e, and TCS1f, were isolated from specific germplasms, and all of recombinant proteins have higher caffeine synthase (CS, EC 2.1.1.160) activity than theobromine synthase (TS, EC 2.1.1.159). Amino acid residue 269 is responsible for the difference in TCS activity and substrate recognition, which was demonstrated by using site-directed mutagenesis experiments. Furthermore, natural variations in TCS1 change the transcription levels. There are two molecular mechanisms controlling the caffeine biosynthesis in low-caffeine-accumulating tea germplasms, i.e., TCS1 allele with low transcription level or its encoded protein with only TS activity. Allelic variations of TCS1 play a crucial role in caffeine biosynthesis. Taken together, our work provides valuable foundation for a comprehensive understanding of the mechanism of caffeine biosynthesis in section Thea plants and useful guidance for effective breeding. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  18. Transcriptomic analysis of Camellia ptilophylla and identification of genes associated with flavonoid and caffeine biosynthesis.

    Science.gov (United States)

    Li, M M; Xue, J Y; Wen, Y L; Guo, H S; Sun, X Q; Zhang, Y M; Hang, Y Y

    2015-12-29

    Camellia ptilophylla, or cocoa tea, is naturally decaffeinated and its predominant catechins and purine alkaloids are trans-catechins and theobromine Regular tea [Camellia sinensis (L.) O. Ktze.] is evolutionarily close to cocoa tea and produces cis-catechins and caffeine. Here, the transcriptome of C. ptilophylla was sequenced using the 101-bp paired-end technique. The quality of the raw data was assessed to yield 70,227,953 cleaned reads totaling 7.09 Gbp, which were assembled de novo into 56,695 unique transcripts and then clustered into 44,749 unigenes. In catechin biosynthesis, leucoanthocyanidin reductase (LAR) catalyzes the transition of leucoanthocyanidin to trans-catechins, while anthocyanidin synthase (ANS) and anthocyanidin reductase (ANR) catalyze cis-catechin production. Our data demonstrate that two LAR genes (CpLAR1 and CpLAR2) by C. ptilophylla may be advantageous due to the combined effects of this quantitative trait, permitting increased leucoanthocyanidin consumption for the synthesis of trans-catechins. In contrast, the only ANS gene observed in C. sinensis (CsANS) shared high identity (99.2%) to one homolog from C. ptilophylla (CpANS1), but lower identity (~80%) to another (CpANS2). We hypothesized that the diverged CpANS2 might have lost its ability to synthesize cis-catechins. C. ptilophylla and C. sinensis each contain two copies of ANR, which share high identity and may share the same function. Transcriptomic sequencing captured two N-methyl nucleosidase genes named NMT1 and NMT2. NMT2 was highly identical to three orthologous genes TCS2, PCS2, and ICS2, which did not undergo methylation in vitro; in contrast, NMT1 was less identical to TCS, PCS and ICS, indicating that NMT1 may undergo neofunctionalization.

  19. Psychostimulant pharmacological profile of paraxanthine, the main metabolite of caffeine in humans

    Science.gov (United States)

    Orrú, Marco; Guitart, Xavier; Karcz-Kubicha, Marzena; Solinas, Marcello; Justinova, Zuzana; Barodia, Sandeep Kumar; Zanoveli, Janaina; Cortes, Antoni; Lluis, Carme; Casado, Vicent; Moeller, F. Gerard; Ferré, Sergi

    2013-01-01

    Caffeine induces locomotor activation by its ability to block adenosine receptors. Caffeine is metabolized to several methylxanthines, with paraxanthine being the main metabolite in humans. In this study we show that in rats paraxanthine has a stronger locomotor activating effect than caffeine or the two other main metabolites of caffeine, theophylline and theobromine. As previously described for caffeine, the locomotor activating doses of paraxanthine more efficiently counteract the locomotor depressant effects of an adenosine A1 than an adenosine A2A receptor agonist. In drug discrimination experiments in rats trained to discriminate a maximal locomotor activating dose of caffeine, paraxanthine, unlike theophylline, generalized poorly to caffeine suggesting the existence of additional mechanisms other than adenosine antagonism in the behavioral effects of paraxanthine. Pretreatment with the nitric oxide inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) reduced the locomotor activating effects of paraxanthine, but not caffeine. On the other hand, pretreatment with the selective cGMP-preferring phosphodiesterase PDE9 inhibitor BAY 73-6691, increased locomotor activity induced by caffeine, but not paraxanthine. Ex vivo experiments demonstrated that paraxanthine, but not caffeine, can induce cGMP accumulation in the rat striatum. Finally, in vivo microdialysis experiments showed that paraxanthine, but not caffeine, significantly increases extracellular levels of dopamine in the dorsolateral striatum, which was blocked by L-NAME. These findings indicate that inhibition of cGMP-preferring PDE is involved in the locomotor activating effects of the acute administration of paraxanthine. The present results demonstrate a unique psychostimulant profile of paraxanthine, which might contribute to the reinforcing effects of caffeine in humans. PMID:23261866

  20. Theophylline detection in serum using a self-assembling RNA aptamer-based gold nanoparticle sensor.

    Science.gov (United States)

    Jiang, Hongyan; Ling, Kai; Tao, Xiaojun; Zhang, Qiqing

    2015-08-15

    Recently, DNA aptamer-gold nanoparticle (AuNP) conjugates have emerged as novel biosensing tools. Although RNA aptamers are more advantageous than DNA aptamers, their vulnerable nature during the construction of these conjugates restricts the development of RNA aptasensors. In this study, we developed an RNA aptamer-based AuNP sensor for the detection of theophylline in serum, combining the high binding affinity and selectivity of a theophylline RNA aptamer and the fluorescence quenching ability of AuNPs. In order to prevent nuclease degradation during the experimental process, the single strand of the theophylline RNA aptamer (33-mer) was split at the end loop region into two shorter halves, which were able to reassemble to form the theophylline-binding pocket. One fragment was linked to a DNA sequence that included a 15 thymine (T15) spacer and a polyadenine (polyA, A12) tail. The chimeric RNA/DNA oligonucleotide was attached to AuNPs within a few minutes via adsorption of the polyA tail. The other fragment was labeled with a fluorophore (Cy3). The two individual fragments self-assembled in the presence of theophylline. Upon ligand binding, the fragments came into close proximity, resulting in fluorescence quenching. This sensor exhibited a low detection limit of 0.05 µM, with a linear dynamic range from 0.1 to 10 µM in serum. Moreover, the sensor did not recognize theophylline-related compounds (e.g., caffeine and theobromine), demonstrating its high selectivity. This strategy offers new possibilities for the application of RNA aptasensors in clinical settings. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Use of Resting Cells of Native Screened Rhodotorula sp. CW03 in Biotransformation of Caffeine to Theophylline and Paraxanthine

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    M. Ashengroph

    2015-07-01

    Full Text Available Introduction & Objective: In recent years, microorganisms have been applied as biocatalysts for making pharmaceutically natural products. Microbial biotransformation of caffeine suggests a dual approach for biodegradation of toxic caffeine from polluted environments and a method for the production of medically and pharmaceutically valuable dimethylxanthines. The present work describes the identification of native yeasts capable of biotransformation of caffeine into theophylline and paraxanthine. Materials & Methods: In this experimental study fourteen yeast strains which were able to de-grade caffeine isolated based on their morphology were selected as biocatalysts for biotrans-formation of caffeine as a low-cost substrate to high value added dimethylxanthines such as theophylline and theobromine. The selected strains were characterized based on phenotypic and genetic tests. Screening was performed by Thin Layer Chromatography (TLC and High Performance Liquid Chromatography (HPLC analyses. Results: The results obtained using TLC and HPLC analyses suggest formation of two main metabolites of theophylline and paraxanthine from biotransformation of caffeine under resting cells of Rhodotorula sp. CW03 (GenBank accession number KF414531. The results showed that under resting cell conditions a maximum concentration of theophylline 380 mg/l (molar yield of 16.4% and paraxanthine 880 mg/l (molar yield of 37.9% were obtained after 72 h and 120 h of conversion time, respectively. Conclusion: In the current investigation, done for the first time in Iran, we describe the isola-tion and identification of yeast strains with caffeine degradation ability which can be proposed as safe and cost-effective biocatalysts in production of value added dimethylxanthines from caffeine as a low-cost substrate.(Sci J Hamadan Univ Med Sci 2015; 22 (2: 83-92

  2. Effects of Paraxanthine and Caffeine on Sleep, Locomotor Activity, and Body Temperature in Orexin/Ataxin-3 Transgenic Narcoleptic Mice

    Science.gov (United States)

    Okuro, Masashi; Fujiki, Nobuhiro; Kotorii, Nozomu; Ishimaru, Yuji; Sokoloff, Pierre; Nishino, Seiji

    2010-01-01

    Study Objective: Caffeine, an adenosine A1 and A2a receptor antagonist, is a widely consumed stimulant and also used for the treatment of hypersomnia; however, the wake-promoting potency of caffeine is often not strong enough, and high doses may induce side effects. Caffeine is metabolized to paraxanthine, theobromine, and theophylline. Paraxanthine is a central nervous stimulant and exhibits higher potency at A1 and A2 receptors, but has lower toxicity and lesser anxiogenic effects than caffeine. Design: We evaluated the wake-promoting efficacy of paraxanthine, caffeine, and a reference wake-promoting compound, modafinil, in a mice model of narcolepsy, a prototypical disease model of hypersomnia. Orexin/ataxin-3 transgenic (TG) and wild-type (WT) mice were subjected to oral administration (at ZT 2 and ZT14) of 3 doses of paraxanthine, caffeine, modafinil, or vehicle. Results: Paraxanthine, caffeine, and modafinil significantly promoted wakefulness in both WT and narcoleptic TG mice and proportionally reduced NREM and REM sleep in both genotypes. The wake-promoting potency of 100 mg/kg p.o. of paraxanthine during the light period administration roughly corresponds to that of 200 mg/kg p.o. of modafinil. The wake-promoting potency of paraxanthine is greater and longer lasting than that of the equimolar concentration of caffeine, when the drugs were administered during the light period. The wake-promotion by paraxanthine, caffeine, and modafinil are associated with an increase in locomotor activity and body temperature. However, the higher doses of caffeine and modafinil, but not paraxanthine, induced hypothermia and reduced locomotor activity, thereby confirming the lower toxicity of paraxanthine. Behavioral evaluations of anxiety levels in WT mice revealed that paraxanthine induced less anxiety than caffeine did. Conclusions: Because it is also reported to provide neuroprotection, paraxanthine may be a better wake-promoting agent for hypersomnia associated with

  3. Semi-volatile compounds variation among Brazilian populations of Ilex paraguariensis St. Hil.

    Directory of Open Access Journals (Sweden)

    Rogério Luis Cansian

    2008-02-01

    Full Text Available The use of compressed carbon towards extracting semi-volatile compounds present in maté leaves (Ilex paraguariensis St. Hil. is due to the growing interest in mate constituents to develop new products in the cosmetic, pharmaceutical and food industries. The objective of this work was to assess the chemical distribution of semi-volatile compounds in 20 native populations of maté collected all over Brazil. The extracts of bulk samples (30 plants of each population were obtained by the high-pressure carbon dioxide extraction technique, and analyzed by GC/MSD. The quantification of compounds (caffeine, theobromine, phytol, squalene, vitamin E, eicosane, pentatriacontane, and stigmasterol showed significant variations within the different populations and compounds analyzed, which are not related to geographical origin or macroclimate characteristics. The results pointed out to the importance of genetic and local environmental factors on the chemical composition of this species.A extração empregando dióxido de carbono a altas pressões em erva-mate (Ilex paraguariensis St. Hil., tem sido justificada pelo crescente interesse desta matriz vegetal ou de parte de seus constituintes na formulação de novos produtos, tais como cosméticos e medicamentos, entre outros. Neste contexto, o objetivo do presente trabalho é avaliar a distribuição química de compostos semi-voláteis em 20 populações de erva-mate coletadas em toda área de distribuição desta espécie no Brasil. 30 plantas foram selecionadas para produzir a amostra de cada população. Os extratos de cada amostra foram obtidos por extração com dióxido de carbono a alta pressão e, posteriormente, foram analisados por CG/EM. A quantificação de alguns compostos semi-voláteis presentes nos extratos (cafeína, teobromina, fitol, esqualeno, vitamina E, eicosano, pentatriacontano e stigmasterol, apresentaram variações significativas entre as concentrações dos diferentes compostos

  4. Determinação de taninos e metilxantinas no guaraná em pó (Paullinia cupana Kunth, Sapindaceae por cromatografia líquida de alta eficiência Determination of tannins and methylxanthines in powdered guarana (Paullinia cupana Kunth, Sapindaceae by high performance liquid chromatography

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    Sandra A. Sousa

    2010-12-01

    Full Text Available As sementes de guaraná (Paullinia cupana Kunth, Sapindaceae são utilizadas como estimulante e os principais constituintes químicos são as metilxantinas e os taninos, tais como catequina e epicatequina. Este trabalho teve por objetivo investigar um método por cromatografia líquida de alta eficiência (CLAE para quantificar, simultaneamente, cafeína, teofilina, teobromina, catequina e epicatequina em pó de guaraná. Os valores obtidos foram comparados com os resultados encontrados no método espectrofotométrico da Farmacopéia Brasileira IV ed. Pelo método cromatográfico, os teores de metilxantinas e taninos totais foram de 4,04% e 1,48%, respectivamente. Os teores de metilxantinas e taninos totais, obtidos pelo método espectrofotométrico, foram de 4,88% e 4,05%, respectivamente. Acredita-se que, nas condições experimentais empregadas, o método espectrofotométrico não foi específico, quantificando outras substâncias fenólicas presentes no guaraná. Portanto, sugere-se o emprego desse método cromatográfico no controle de qualidade do guaraná em pó, por ser preciso, sensível, específico e rápido.The seeds of Paullinia cupana Kunth, Sapindaceae, are used as a stimulant and the main chemical constituents are the methylxanthines and tannins, as catechin and epicatechin. This study aimed to investigate a method by high performance liquid chromatography (HPLC to quantify both caffeine, theophylline, theobromine, catechin and epicatechin in powdered of P. cupana. The values obtained were compared with the results of the spectrophotometric method of the Brazilian Pharmacopoeia IV ed. For the chromatographic method, the levels of methylxanthines and total tannins were 4.04% and 1.48%, respectively. The levels of methylxanthines and total tannins obtained by the spectrophotometric method, were 4.88% and 4.05%, respectively. It is believed that, under our experimental conditions, the spectrophotometric method was not specific and

  5. Validated high-performance liquid chromatographic method for the standardisation of Ptychopetalum olacoides Benth., Olacaceae, commercial extracts Validação de método de cromatografia líquida de alta performance para padronização de extratos comerciais de Ptychopetalum olacoides Benth., Olacaceae

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    Renata Colombo

    2010-11-01

    Full Text Available Ptychopetalum olacoides Benth., Olacaceae, popularly known as marapuama or muirapuama or miriantã, is a species native to the Amazonian region of Brazil. Extracts of the bark of the plant have been used traditionally for its stimulating and aphrodisiac properties and currently commercialised by the herbal industry as constituents in a wide range of phytomedicines. Fractionation by open column chromatography followed by preparative HPLC-UV/PAD of the stem bark and of three commercial extracts of P. olacoides allowed the isolation of three components that were common to all extracts analysed, and these were identified by NMR to be vanillic acid, protocatechuic acid and theobromine. Vanillic acid, which has been proposed as a phytochemical marker for P. olacoides, was employed as an external standard in the development and validation of a rapid qualitative and quantitative HPLC assay for the analyte. The recoveries values of the developed method were 99.02% and the LOD and LOQ values were 0.033 and 0.11 mg.L-1, respectively. The described method may be applied to the standardisation of herbs, extracts or phytomedicines commercialised as marapuama.Ptychopetalum olacoides Benth., Olacaceae, popularmente conhecida como marapuama, muirapuama ou miriantã, é uma espécie nativa da região da Amazônia do Brasil. Extratos das cascas da planta são tradicionalmente usados por suas propriedades estimulantes e afrodisíacas, e frequentemente comercializados como constituinte de uma grande variedade de formulações fitoterápicas. O fracionamento por coluna cromatográfica aberta seguida por CLAE-UV/PAD das cascas do caule de três extratos comerciais de P. olacoides permitiram o isolamento de três substâncias comuns em todos os extratos analisados. Os compostos foram identificados por RMN como ácido vanílico, ácido protocatecuíco e teobromina. O ácido vanílico foi utilizado como marcador fitoquímico para P. olacoides e empregado como padr

  6. Dietary nutrients associated with short and long sleep duration. Data from a nationally representative sample.

    Science.gov (United States)

    Grandner, Michael A; Jackson, Nicholas; Gerstner, Jason R; Knutson, Kristen L

    2013-05-01

    Short sleep duration is associated with weight gain and obesity, diabetes, cardiovascular disease, psychiatric illness, and performance deficits. Likewise, long sleep duration is also associated with poor physical and mental health. The role of a healthy diet in habitual sleep duration represents a largely unexplored pathway linking sleep and health. This study evaluated associations between habitual sleep parameters and dietary/nutritional variables obtained via the National Health and Nutrition Examination Survey (NHANES), 2007-2008. We hypothesized that habitual very short (short (5-6h) and long (9+h) sleep durations are associated with intake of a number of dietary nutrient variables. Overall, energy intake varied across very short (2036kcal), short (2201kcal), and long (1926kcal) sleep duration, relative to normal (2151kcal) sleep duration (p=0.001). Normal sleep duration was associated with the greatest food variety (17.8), compared to very short (14.0), short (16.5) and long (16.3) sleep duration (psleep duration were found across nutrient categories, with significant associations between habitual sleep duration and proteins, carbohydrates, vitamins and minerals. In stepwise analyses, significant contributors of unique variance included theobromine (long sleep RR=0.910, pshort sleep RR=0.890, pshort sleep RR=0.952, pshort (sleep RR=0.941, pshort sleep RR=1.123, psleep RR=0.812, psleep RR=0.450, p=0.001), lycopene (very short (sleep RR=0.950, pshort (sleep RR=0.494, psleep RR=0.509, pshort sleep RR=0.670, psleep RR=1.172, pshort and/or long sleep duration, which may be explained by differences in food variety. Future studies should assess whether these associations are due to appetite dysregulation, due to short/long sleep and/or whether these nutrients have physiologic effects on sleep regulation. In addition, these data may help us better understand the complex relationship between diet and sleep and the potential role of diet in the relationship between

  7. Nutritional and Phytochemical Study of Ilex paraguariensis Fruits

    Directory of Open Access Journals (Sweden)

    Laura Cogoi

    2013-01-01

    Full Text Available Yerba mate is defined as the product constituted by the dried, slightly roasted, and milled leaves of Ilex paraguariensis. However, the fruits of this species are often found in the commercial product. Nowadays the fruits are considered a byproduct. The objective of this work was to obtain the preliminary data of minerals, lipids, methylxanthines and polyphenols in the ripe fruits of I. paraguariensis. The results showed a considerable amount of total dietary fiber (42.0 ± 1.6 g/100 g and nutritionally valuable minerals: potassium (1324 ± 15 mg/100 g, iron (6.4 ± 0.5 mg/100 g, magnesium (168 ± 15 mg/100 g, calcium (150 ± 12 mg/100 g, copper (1.1 ± 0.1 mg/100 g, zinc (2.3 ± 0.3 mg/100 g, and sodium (1.3 ± 0.1 mg/100 g. The lipid content was 4.5 g/100 g. Oleic acid was the predominant unsaturated fatty acid (38.74 ± 0.75 g/100 g. Linoleic acid (1.83 ± 0,01 g/100 g was also present. Methylxanthines were quantified: caffeine (0.118 ± 0.001 and theobromine (0.0125 ± 0.0002 g/100 g. The total polyphenol content was 0.717 ± 0.001 g/100 g. The results obtained in this work suggest the potential value of the fruits of I. paraguariensis for the development of novel products in the food and pharmaceutical industries. This paper aims to contribute to the scientific knowledge of a natural by-product from industry regarding the need of foods and medicines for the new millennium.

  8. Resonance Enhanced Multi-photon Spectroscopy of DNA

    Science.gov (United States)

    Ligare, Marshall Robert

    its intrinsic molecular properties and possibly a hydrogen bonding solvent capable of dissipating excess vibrational energy. Due to the high specificity and sensitivity of resonant two photon ionization coupled with molecular beam mass spectrometry a new analytical technique for identifying molecular markers in archaeological vessels is presented. The xanthine alkaloids theobromine, theophylline and caffeine are identified in Central American and North American pottery sherds by direct desorption/resonant laser ionization mass spectrometry.

  9. Adsorption behavior of caffeine as a green corrosion inhibitor for copper

    Energy Technology Data Exchange (ETDEWEB)

    Souza, Fernando Silvio de [Grupo de Estudos de Processos Eletroquimicos e Eletroanaliticos, Departamento de Quimica, Universidade Federal de Santa Catarina, 88040-900, Florianopolis, SC (Brazil); Giacomelli, Cristiano [Departamento de Quimica, Universidade Federal de Santa Maria, Av. Roraima 1000, 97119-900, Santa Maria, RS (Brazil); Goncalves, Reinaldo Simoes [Universidade Federal do Rio Grande do Sul, Av. Bento Goncalves 9500, 91501-970, Porto Alegre, RS (Brazil); Spinelli, Almir, E-mail: almir.spinelli@ufsc.br [Grupo de Estudos de Processos Eletroquimicos e Eletroanaliticos, Departamento de Quimica, Universidade Federal de Santa Catarina, 88040-900, Florianopolis, SC (Brazil)

    2012-12-01

    Electrochemical and impedance experiments were carried out to evaluate the corrosion behavior of copper in aerated 0.1 mol L{sup -1} H{sub 2}SO{sub 4} solutions in the presence of three xanthine derivatives with similar chemical structures. The corrosion rate of copper was found to increase in the presence of theophylline and theobromine and decrease in the presence of caffeine. The adsorption and inhibitory effect of caffeine on copper surfaces in aerated 0.1 mol L{sup -1} H{sub 2}SO{sub 4} solutions were then investigated in detail by potentiodynamic polarization, electrochemical impedance spectroscopy (EIS), contact angle measurements, scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS) and fluorescence experiments. The data obtained indicate that caffeine behaves as a cathodic-type inhibitor adsorbing onto the copper surface according to the Temkin isotherm, with the negative Increment G Degree-Sign {sub ads} value of - 31.1 kJ mol{sup -1} signifying a spontaneous adsorption process. The corrosion inhibition efficiency increased with caffeine concentration in the range of 1.0-10.0 mmol L{sup -1}. Furthermore, the EIS results obtained at the open-circuit potential and surface analysis (SEM, EDS and fluorescence) clearly demonstrated the adsorption of the organic compound onto the copper electrode. The contact angle measurements revealed the formation of a hydrophobic protective film. This film covers up to 72% of the total active surface, acts as a protective barrier and prevents interaction between the metal, water and oxygen molecules. - Highlights: Black-Right-Pointing-Pointer We have investigated the adsorption and corrosion inhibition of caffeine on copper surfaces. Black-Right-Pointing-Pointer Caffeine behaves as a cathodic-type inhibitor. Black-Right-Pointing-Pointer Caffeine adsorbs onto copper surface according to Temkin isotherm. Black-Right-Pointing-Pointer There exists the formation of a hydrophobic film that acts as a

  10. Complexes of poly(adenylic acid) with complementary monomers.

    Science.gov (United States)

    Davies, R J

    1976-01-02

    The interaction of a number of potentially complementary monomers with poly(A) has been investigated by equilibrium dialysis, optical rotatory dispersion and ultraviolet absorption measurements. Experiments were conducted at pH 7.0, 0.15 M Na+, where poly(A) exists as a random coil with some degree of base-stacking, and at pH 6.0, 0.15 M Na+, where poly(A) adopts the protonated double-helical acid form structure below 15 degrees C. Binding isotherms show that, at 3.5 degrees C, poly(A) forms a 1 : 1 complex with xanthine at pH 6, and a 2:1 complex at pH 7, while oxoformycin forms a 1:1 complex with poly(A) at both pH 6 and pH 7. Poly(A) forms a complex, tentatively assigned 1:1 stoichiometry, with 8-azaxanthine at pH 6, but no complexing occurs at pH 7. The complexes have been characterized by their optical rotatory dispersion and ultraviolet spectra, their thermal stabilities, and their rates of formation at low temperature. All the complexes are laevorotatory at long wavelengths (greater than 300 nm) and unplex formation at low temperature is a slow process requiring many hours for completion. The complexes of poly(A) with 3-methylxanthine have been reinvestigated and shown to undergo normal helix-coil transitions; the anomalous melting behaviour noted previously [Biopolymers, 10, 21 -- 33 (1971)] has been explained. From a comparison of optical rotatory dispersion spectra, it is concluded that the poly(A) with 3-methylxanthine have similar structures, which are quite different from the structures of the corresponding complexes with 7-methylxanthine. The structures and properties of the poly(A) - monomer complexes are discussed, and compared with those of other polynucleotide - monomer complexes. No significant interaction was observed between poly(A) and hypoxanthine, allopurinol, 6,8-dihydroxypurine, 1-methylxanthine, 9-methylxanthine, theophylline, theobromine or 3,9-dimethylxanthine.

  11. Ilex paraguariensis Promotes Orofacial Pain Relief After Formalin Injection: Involvement of Noradrenergic Pathway.

    Science.gov (United States)

    de Carvalho, Eudislaine Fonseca; de Oliveira, Simone Kobe; Nardi, Viviane Koepp; Gelinski, Tathiana Carla; Bortoluzzi, Marcelo Carlos; Maraschin, Marcelo; Nardi, Geisson Marcos

    2016-03-01

    Drinking mate or chimarrão, a hot infusion of Ilex paraguariensis (ILEX) leaves, is a common habit in Southern South America that has a social and almost ritualistic role. It has been used as a stimulant beverage in South America and analgesic in regions of Argentina for treatment of headache and others painful inflammatory conditions such as arthritis and rheumatism. The aim of this study was to evaluate the pharmacological activity of I. paraguariensis infusion (ILEX) on orofacial nociception model induced by formalin, and study its mechanism of action. The analgesic effect of ILEX was assessed through writhing test, paw formalin test, paw edema induced by carrageenan, and orofacial pain induced by formalin. To study the action mechanism of ILEX, opioidergic, dopaminergic, nitrergic, and adrenergic pathways were investigated. The high-performance liquid chromatography analysis of ILEX infusion revealed caffeine and theobromine. The treatment with ILEX reduced the number of writhing. However, it was effective neither in the formalin paw test nor in the paw edema induced by carrageenan. Different from formalin paw test, ILEX was able to reduce the orofacial reactivity to formalin in 31.8% (70.4 ± 2.5 s; first phase), and 20% (127.3 ± 18.9 s; second phase). The analgesic effect of ILEX results from the modulation of noradrenergic pathways since prazosin (α1-adrenoceptor antagonist, 0.15 mg/kg; intraperitoneal) reversed the analgesic effect of ILEX. The present report demonstrates that analgesic effect of ILEX in orofacial formalin test is due mainly to modulation of noradrenergic pathways. Ilex paraguariensis (ILEX) has been used as a stimulant beverage in South America and analgesic in regions of Argentina for the treatment of headache and others painful inflammatory conditions such arthritis and rheumatism.The aim of this study was to evaluate the pharmacological activity of ILEX on orofacial nociception model induced by formalin, and study its mechanism of

  12. PILOT STUDY: An international comparison of mass fraction purity assignment of theophylline: CCQM Pilot Study CCQM-P20.e (Theophylline)

    Science.gov (United States)

    Westwood, S.; Josephs, R.; Daireaux, A.; Wielgosz, R.; Davies, S.; Kang, M.; Ting, H.; Phillip, R.; Malz, F.; Shimizu, Y.; Frias, E.; Pérez, M.; Apps, P.; Fernandes-Whaley, M.; DeVos, B.; Wiangnon, K.; Ruangrittinon, N.; Wood, S.; Duewer, D.; Schantz, M.; Bedner, M.; Hancock, D.; Esker, J.

    2009-01-01

    Under the auspices of the Organic Analysis Working Group (OAWG) of the Comité Consultatif pour la Quantité de Matière (CCQM) a laboratory comparison, CCQM-P20.e, was coordinated by the Bureau International de Poids et Mesures (BIPM) in 2006/2007. Nine national measurement institutes, two expert laboratories and the BIPM participated in the comparison. Participants were required to assign the mass fraction of theophylline present as the main component in two separate study samples (CCQM-P20.e.1 and CCQM-P20.e.2). CCQM-P20.e.1 consisted of a high-purity theophylline material obtained from a commercial supplier. CCQM-P20.e.2 consisted of theophylline to which known amounts of the related structure compounds theobromine and caffeine were added in a homogenous, gravimetrically controlled fashion. For the CCQM-P20.e.2 sample it was possible to estimate gravimetric reference values both for the main component and for the two spiked impurities. In addition to assigning the mass fraction content of theophylline for both materials, participants were requested but not obliged to provide mass fraction estimates for the minor components they identified in each sample. The results reported by the study participants for the mass fraction content of theophylline in both materials showed good levels of agreement both with each other and with the gravimetric reference value assigned to the CCQM-P20.e.2 material. There was also satisfactory agreement overall, albeit at higher levels of uncertainty, in the quantification data reported for the minor components present in both samples. In the few cases where a significant deviation was observed from the consensus values reported by the comparison participants or gravimetric reference values where these where available, they appeared to arise from the use of non-optimal chromatographic separation conditions. The results demonstrate the feasibility for laboratories to assign mass fraction content with associated absolute expanded

  13. NTP Toxicology and Carcinogenesis Studies of Theophylline (CAS No. 58-55-9) in F344/N Rats and B6C3F1 Mice (Feed and Gavage Studies).

    Science.gov (United States)

    1998-08-01

    Theophylline is an alkaloid found in tea (Thea sinensis) and chocolate and is structurally related to caffeine and theobromine. Theophylline is used as a pharmaceutical agent. It stimulates the heart and central nervous system, relaxes the smooth muscles of the bronchi and blood vessels, and causes diuresis. The drug is used mainly as a bronchodilator in obstructive airway diseases, such as bronchial asthma, and for myocardial stimulation. Theophylline was nominated for toxicologic and carcinogenicity testing as a representative of the purine structural subclass, particularly because of its relationship to purines such as caffeine, 1-methyl-3-hydroxyguanine, and 3-hydroxy-1-methylxanthine, the latter two compounds having been shown to induce sarcomas in rats. Additional reasons for testing theophylline included its widespread use in humans as a pharmaceutical agent, its possible genotoxicity in vitro, and the lack of information on its potential toxicity and/or carcinogenicity under conditions of chronic oral usage. Based on reported teratogenicity and testicular toxicity, it was also recommended that reproductive studies be included in the evaluation of theophylline. The oral route of administration was selected because it is the primary route of human exposure, and the gavage route was selected because it mimics the pharmaceutical use of theophylline in humans. Male and female F344/N rats and B6C3F1 mice were given theophylline (greater than 99% pure) in feed or in corn oil by gavage for 16 days or 14 weeks or in corn oil by gavage for 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, mouse bone marrow, and mouse peripheral blood. 16-DAY FEED STUDY IN RATS: Groups of five male and five female F344/N rats were given 0, 500, 1,000, 2,000, 4,000, or 8,000 ppm theophylline in feed for 16 days, which resulted in approximate daily doses of 50, 100, 250, 450, or 1,000 mg theophylline/kg body weight to