Actividad antioxidante en genotipos de Theobroma spp. (Malvaceae) en México / Antioxidant activity in genotypes of Theobroma spp. (Malvaceae) in Mexico

Abstract Introduction: The organoleptic qualities of aromatic species and their derived products are directly related to some characteristics of flavor, color and nutritional value and depend largely on their genetic origin and content of secondary metabolites. Objective: The antioxidant activity of different genotypes of Theobroma spp. from Mexico was evaluated in order to distinguish promising qualities for genetic improvement, and to differentiate phylogenetic traits, considering biochemical variables. Methods: The amount of phenols, flavonoids and antioxidant activity was determined by ABTS and DDPH, in addition to the content of anthocyanins, theobromine and caffeine in four species of Theobroma L., and 50 genotypes derived from T. cacao. The results were analyzed using an analysis of variance, means test, principal component analysis and cladistic analysis. Results: There are highly significant differences between genotypes. The phenol content ranged from 7.5-85 mg g-1; flavonoids 6.57-69.6 mg g-1, antioxidant activity by ABTS of 17.3-86.1 and by DDPH of 40.0-53.3; anthocyanin content of 0.01-3, caffeine of 1.8-6.7-and theobromine of 2.9-9.8 mg g-1. Principal component and cladistic analysis helped explain the variation found and distinguish evolutionary characters and phylogenetic brotherhoods. The variation in content of phenols, flavonoids, antioxidant activity, anthocyanins, theobromine and caffeine was mainly due to the degree of domestication, while for the group of genotypes derived from T. cacao (forastero, trinitario and criollo) it was the origin of the seeds. Conclusions: The degree of domestication influences the content of phenols and antioxidant capacity. The results suggest that the evaluated variables can help to form criteria for genetic improvement in the complex derived from T. cacao oriented to the selection of higher phenol content and greater antioxidant activity.

Resumen Introducción: Las cualidades organolépticas de las especies aromáticas y sus productos derivados se relacionan directamente con algunas características del sabor, color y valor nutricional y dependen en gran medida de su origen genético y contenido de metabolitos secundarios. Objetivo: Se evaluó la actividad antioxidante de diferentes genotipos de Theobroma spp. de México, con el fin de distinguir cualidades promisorias para el mejoramiento genético, y diferenciar rasgos filogenéticos, considerando variables bioquímicas. Métodos: Se determinó la cantidad de fenoles, flavonoides y actividad antioxidante mediante ABTS y DDPH, además de contenido de antocianinas, teobromina y cafeína en cuatro especies de Theobroma L., y 50 genotipos derivados de T. cacao. Resultados: Los resultados fueron analizados mediante un análisis de varianza, prueba de medias, análisis de componentes principales y análisis cladístico. Existen diferencias altamente significativas entre genotipos. El contenido de fenoles varió de 7.5-85 mg g-1; flavonoides 6.57-69.6-mg g-1, actividad antioxidante por ABTS de 17.3-86.1 y por DDPH de 40.0-53.3; el contenido de antocianinas de 0.01-3, cafeína de 1.8-6.7 y teobromina de 2.9-9.8 mg g-1. El análisis de componentes principales y cladístico ayudó a explicar la variación encontrada y distinguir caracteres evolutivos y hermandades filogenéticas. La variación en contenido de fenoles, flavonoides, actividad antioxidante, antocianinas, teobromina y cafeína estuvo dada principalmente por el grado de domesticación, mientras que para el grupo de genotipos derivados de T. cacao (forastero, trinitario y criollo) fue el origen de las semillas. Conclusión: El grado de domesticación influye en el contenido de fenoles y actividad antioxidante. Los resultados sugieren que las variables evaluadas pueden ayudar a formar criterios para el mejoramiento genético en el complejo derivado de T. cacao orientado a la selección de mayor contenido de fenoles y mayor actividad antioxidante.

Assessment of remineralization potential of Theobromine and Sodium Fluoride gels on Artificial Caries like lesions / Avaliação do potencial de remineralização de géis de teobromina e fluoreto de sódio em lesões de cárie artificial

Objective: To compare the remineralization potential of theobromine and sodium fluoride gels on artificial caries like lesion. Materials and Methods: Forty longitudinal halves of human mandibular premolars were equally divided into 4 groups of 10 samples each: control group (C), samples were stored in distilled water during the experiment period. The remaining 30 specimens were subjected to demineralization protocol to create caries like lesions. samples were immersed for three days in a demineralization solution (pH 5.0) containing 0.2% carbopol and 0.1% lactic acid saturated with calcium phosphate tribasic. The samples were subdivided into 3 equal groups according to the treatment applied during the pH cycle. Demineralization group "D": no treatment applied. Group "F" treated with 2000 mg/liter sodium fluoride gel. Group "T" treated with 200 mg/liter theobromine gel. The specimens of the two studies groups were subjected to Demineralization- Remineralization PH Cycle Protocol for 5 days (Alternating four steps: 1: Treatment material, fluoride or theobromine ˜= 3 minutes. 2: Demineralizing solution 3 hours. 3: treatment material ˜= 3 minutes. 4: Remineralizing solution till the next cycle). The samples were investigated by scanning electron microscope (SEM) and energy dispersive x-ray analysis (EDXA). Results: The enamel of the demineralization group was porous with artificial caries like changes exposing the subsurface enamel rods with severe rod core defects. Theobromine gel and fluoride gel groups showed marked improvement in the surface characteristics in the enamel in both groups. Theobromine gel group showed more observable enamel surface improvement than the fluoride gel group. EDXA revealed that the calcium-phosphorus ratio displayed a descending order: (C > T > F > D). The differences between the two tested groups were not statistically significant. Conclusion: Theobromine gel had more effective remineralizing potential than fluoride gel as a result of its effect in improving the enamel surface characteristics of human teeth. (AU)

Objetivo: Comparar o potencial de remineralização dos géis de teobromina e fluoreto de sódio em lesões de cáries artificiais. Materiais e Métodos: Quarenta metades longitudinais de pré-molares inferiores humanos foram igualmente divididas em 4 grupos de 10 amostras cada: grupo controle (C), as amostras foram armazenadas em água destilada durante o período do experimento. As 30 amostras restantes foram submetidas ao protocolo de desmineralização paracriar lesões artificiais de cárie. As amostras foram imersas por três dias em uma solução de desmineralização (pH 5,0) contendo 0,2% de carbopol e 0,1% de ácido lático saturado com fosfato de cálcio tribásico. As amostras foram subdivididas em 3 grupos iguais, de acordo com o tratamento aplicado durante o ciclo do pH. Grupo de desmineralização "D": nenhum tratamento aplicado. Grupo "F" tratado com 2000 mg / litro de fluoreto de sódio em gel. Grupo & quot;T &q uot; tratado com 200 mg / litro de gel de teobromina. As amostras dos dois grupos de estudo foram submetidas ao protocolo de ciclo de desmineralização - remineralização por 5 dias (quatro etapas alternativas: 1: material de tratamento, flúor ou teobromina ˜= 3 minutos. 2: solução desmineralizante 3 horas. 3: material de tratamento ˜= 3 minutos 4: Solução de remineralização até o próximo ciclo). As amostras foram investigadas por microscopia eletrônica de varredura(MEV) e análise de raios-x dispersivos de energia (EDXA). Resultados: O esmalte do grupo de desmineralização era poroso, com cáries artificiais, como alterações que expunham as hastes de esmalte do subsolo com graves defeitos no núcleo da haste. Os grupos gel de teobromina e flúor apresentaram melhora acentuada nas características da superfície do esmalte nos dois grupos. O grupo gel de teobromina mostrou uma melhoria na superfície do esmalte mais observável do que o grupo gel de fluoreto. A EDXA revelou que a razão cálcio-fósforo exibia uma ordem decrescente: (C> T> F> D). As diferenças entre os dois grupos testados não foram estatisticamente significantes. Conclusão: O gel de teobromina teve um potencial remineralizante mais eficaz que o gel de flúor como resultado de seu efeito na melhoria das características da superfície do esmalte dos dentes humanos. (AU)

Identification of structural analogues xanthine, theophylline and theobromine by surface-enhanced Raman spectroscopy / 药学实践杂志

Objective To distinguish the structural analogues xanthine, theophylline and theobromine by surface-enhanced Raman spectroscopy. Methods Concentrated silver colloid enhancement reagent was prepared as the Raman substrate to increase the number of "hot spots" per unit area, improve the sensitivity of surface-enhanced Raman spectroscopy, enhance the signal strength of the samples and achieve the effective discrimination of structural analogues. Meanwhile, the feasibility of surface-enhanced Raman spectroscopy in practical application was verified by determining serum samples of three mixtures. Results The concentrated silver colloid greatly increased the Raman intensity of the three structural analogues. The spectra of each individual compound and the mixture in the serum system was obtained. The detection limit of the three substances in aqueous solution were 0.005, 0.01 and 0.005 μmol/L respectively. Conclusion Surface-enhanced Raman spectroscopy is a potent technique for distinguishing structural analogues. It is rapid, sensitive and nondestructive to samples. Hence, it can be widely used in the fields of detection, analysis, clinical treatment and diagnosis.

Extracción de compuestos solubles de la cascarilla de cacao con CO2 supercrítico. Caso de metilxantinas y grasa / Extraction of soluble compounds from cocoa shell with supercritical CO2 . A methilxantines and fat case

Resumen La extracción supercrítica es una técnica efectiva para separar diversos compuestos desde matrices naturales, por las propiedades que le confiere a los solventes como el CO2en estado supercrítico. El objetivo del presente trabajo fue la extracción supercrítica de teobromina, cafeína y grasa de la cascarilla de cacao con CO2supercrítico. Se propuso un diseño 22 con 5 puntos centrales, en condiciones estáticas de extracción, teniendo como factores de estudio, presión (2 000 psi a 6 000 psi) y temperatura (318 K a 333 K). Se analizaron los residuos en la cámara de extracción para evaluar los porcentajes de remoción y residual de grasa, cafeína y teobromina, respectivamente. Los resultados mostraron un rendimiento entre 1.72 % a 9.57 %, con una razón de remoción para grasa y cafeína de 61.31 % a 94.54 % y 38.52 % a 78.38 %, respectivamente. La teobromina se retuvo en el polvo de la cascarilla con una razón residual mayor a 90 %. Los efectos de las variables de temperatura y presión para la remoción de grasa fueron predichos a través de un polinomio de primer orden con efectos de interacción, no así para cafeína o teobromina. El método de extracción fue eficiente para remover grasa y cafeína, quedando un residuo en la cámara de extracción considerado como rico en compuestos funcionales.

Abstract Supercritical extraction is an effective technique for separating various compounds from natural matrices due to the properties from solvents, such as CO2, in supercritical state. In the present work, the supercritical extraction of theobromine, caffeine and fat from cocoa shell was carried out with supercritical CO2. A 22 design with 5 central points was proposed, with static operation conditions, having pressure (2 000 psi to 6 000 psi) and extraction temperature (318 K to 333 K) as study factors. The residues were analyzed in the extraction chamber to evaluate the removal and residual percentages of fat, caffeine and theobromine, respectively. The results showed a yield between 1.72 % to 9.57 %, with a removal rate for fat and caffeine of 61.31 % to 94.54 % and 38.52 % to 78.38 %, respectively. The theobromine was retained in the shell powder with a residual ratio greater than 90 %. The effects of temperature and pressure for fat were predicted through a first-order polynomial with interaction effects, but not for caffeine or theobromine. The extraction method was efficient to remove fat and caffeine, leaving a residue powder in the extraction chamber considered as high in functional compounds.

Degradation of caffeine by Pseudomonas monteilii KRM9

@#Aims: The objective of this research was to isolate caffeine-degrading bacteria from coffee pulp waste in Indonesia andcharacterize their caffeine degradation activity.Methodology and results: The caffeine-degrading bacteria were isolated from coffee pulp wastes of Coffea arabicaand C. canephora. These isolates were selected based on their caffeine degradation activity. The identification andbiochemical properties of the best isolate were conducted via 16S rDNA sequence analyses and by using the Microbactkit. Meanwhile, caffeine degradation activity of this bacteria was analyzed by using LC-MS/MS. The results indicatedthat fourteen bacterial isolates were able to degrade caffeine. The highest caffeine degradation activity was performedby isolate KRM9 at the rate of 99.26 ± 0.01%, on a caffeine medium after 24 h of incubation. Based on the 16S rDNAanalyses, the KRM9 isolate was identified as Pseudomonas monteilii. Till present, this species has not been reported asa caffeine-degrading bacterium. However, LC-MS/MS analysis indicated that caffeine was degraded by P. monteiliiKRM9 and theobromine was not the secondary metabolite of caffeine degradation.Conclusion, significance and impact of study: Pseudomonas monteilii KRM9 was detected as a new isolate ofcaffeine-degrading bacteria. This bacterium can be introduced as an agent to degrade caffeine from coffee pulp waste. Itis expected that further research can be conducted on the overall mechanism of caffeine degradation by P. monteiliiKRM9

Efficacy and Tolerability of Theobromine in Patients with Upper Airway Cough Syndrome: A Multicenter, Double-Blind, Randomized, Active Drug Comparative Study

OBJECTIVES: The objective of this study is to evaluate the efficacy and tolerability of theobromine in patients with upper airway cough syndrome compared to those of levocloperastine. MATERIALS AND METHOD: This was a randomized, double-blind study. One hundred sixty-five patients with upper airway cough syndrome participated in a 5 day treatment; 85 subjects were included in the theobromine treatment group and 80 in the levocloperastine control group. Cough severity score, daytime cough symptom (DCS), nighttime cough symptom (NCS) and cough quality of life questionnaire (CQLQ) were analyzed for symptom analysis, and vital signs and laboratory study were performed for safety evaluation before and after medication administration. RESULTS: The primary efficacy analysis showed that the mean change in cough grade between baseline and follow-up in the treatment group was lower than that in the control group. This led to the conclusion that theobromine has similar efficacy to control treatment. The secondary efficacy analysis of changes in DCS, NCS and CQLQ verified this conclusion. Considering safety, only one case of dyspepsia was considered to be probably related to theobromine. Other tests conducted before and after treatment confirmed the safety of treatment medications. CONCLUSION: Theobromine is a novel natural antitussive medication that has similar efficacy to levocloperastine and adequate safety.

Extraction and estimation of theobromine in marketed tea by HPTLC and UV method.

A rapid, simple and sensitive chromatographic method (RP-HPTLC) has been developed for the extraction and quantitative estimation of Theobromine in different extract tea samples (Camellia sinensis). Separation was performed on Silica gel 60 f254 HPTLC plates with ethyl acetate: methanol (27:3 v/v), as mobile phase. The determination was carried out in the UV region, using densitometric absorbance at 274 nm. The maximum concentration of Theobromine in tea samples was found to be 2.313 %. The theobromine response was found to be linear over range 3-15 μg per zone. Limit of detection and quantitation were found to be 30 and 140 ng/ spot respectively. The HPTLC method was validated in terms of precision, accuracy, sensitivity and robustness. The proposed method provided precise and accurate analysis for extraction and estimation of theobromine by HPTLC-UV.

Determination of Theobromine in Compound Theophylline and Ephedrine Tablet by HPLC / 中国中医药信息杂志

Objective To establish a HPLC method for determination of theobromine in Compound Theophylline and Ephedrine Tablet. Method HPLC method was adopted on Kromasil C18 column (250 mm? 4.6 mm, 5 ?m), and temperature was set at 45 ℃. The mobile phase consisted of 0.05 mol/L KH2PO4- Methanol-Triethanolamine (77∶23∶0.2), with flow rate of 0.8 mL/min. The UV detection wavelength was set at 215 nm. Results The calibration curves of theobromine was linear in the range of 0.3 094～0.722 ?g (r=0.999 96), ant the average recovery was 100.49% (n=9) with RSD=1.1%. Conclusion This method is simple, rapid, sensitive and accurate, and can be used for determination of theobromine in Compound Theophylline and Ephedrine Tablet.