Efeito da acidez e de modificadores orgânicos na determinação de metilxantinas: um experimento de cromatografia liquida de alta eficiência (CLAE) empregando otimização uni e multivariada

In this work a new experiment using HPLC is proposed in order to explore the role of acidity and the organic modifiers in the determination of methylxanthines in tea and coffee. Multivariate and univariate optimizations of the experimental conditions were used.

Avaliação da composição mineral do chá da folha senescente de Montrichardia linifera (arruda) schott (araceae) por espectrometria de absorção atômica com chama (FAAS)

Levels of Ca, Mg, Fe, Zn, Cu and Mn were determinated by FAAS in the tea and dry matter from senescent leaf from Montrichardia linifera, plant used in folk medicine Amazon. The content of these metals that are transferred of the leaf to the infusions have presented significant reductions, however, the Mn values in the infusion may exceed the tolerable daily intake (11 mg) if consumption of this tea is greater than 1.0 L per day. So the tea of senescent leaves of M. linifera may be considered as a toxic beverage and thus its use is not advised.

Nanostructured systems containing an essential oil: protection against volatilization

The goal of this study was to evaluate the feasibility of preparing nanocapsules and nanoemulsions using tea tree oil as oily phase aiming to protect its volatilization. The nanostructures presented nanometric mean size (160-220 nm) with a polydispersity index below 0.25 and negative zeta potential. The pH values were 6.43 ± 0.37 and 5.98 ± 0.00 for nanoemulsions and nanocapsules, respectively. The oil content after preparation was 96%. The inclusion of tea tree oil in nanocapsules showed higher protection against volatilization. The analysis of mean size and polydispersity index of formulations presented no significant alteration during the storage time.

Voltammetric determination of sibutramine in beverages and in pharmaceutical formulations

A simple and sensitive method has been proposed for the determination of sibutramine-HCl in energy drinks, green tea and pharmaceutical formulations using differential pulse voltammetry performed on a hanging mercury drop electrode. In the chosen experimental condition (Mcllvaine pH 4.0 buffer, 50 mV pulse amplitude and 40 mV s-1 scan velocity), sibutramine-HCl presented a reversible behavior and a peak maximum at -80 mV. Detection limit was 0.4 mg L-1 and the working linear range extended up to 33.3 mg L-1 (r = 0.99). Analysis of real and fortified samples enabled recoveries between 91 and 102%. The electroanalytical method was compared with a HPLC method which indicated it accuracy.

Extração/pré-concentração de íons cobre no ponto nuvem explorando a formação de complexos com DMIT [4,5-dimercapto-1,3-ditiol-2-tionato]

The present study proposes a method for cloud point preconcentration of copper ions at pH 2.0 based on complexes formed with [4,5-dimercapto-1,3-dithyol-2-thionate] and subsequent determination by flame atomic absorption spectrometry (FAAS). Under optimal analytical conditions, the method provided limits of detection of 0.84 and 0.45 µg L-1, by preconcentrating 12.0 and 24.0 mL of sample, respectively. The method was applied for copper determination in water samples, synthetic saliva, guarana powder, tea samples and soft drinks and the accuracy was assessed by analyzing the certified reference materials Dogfish Liver (DOLT-4) and Lobster Hepatopancreas (TORT-2).

Separation and preconcentration of copper in environmental samples on Amberlite XAD-8 resin after complexation with a carbothioamide derivative

A new solid phase extraction (SPE) method has been developed for the selective separation and preconcentration of Cu (II) ions in food and water samples prior to its flame atomic absorption spectrometry determination. The method is based on the adsorption of the Cu(II) - 2-{[4-Amino-3-(4-methylphenyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-1-yl]acetyl}-N-phenyl hydrazinecarbothioamide complex on Amberlite XAD-8 resin. The metal complex retained on the resin was eluted with 7.5 mL of 2.0 mol L-1 HCl in acetone. The optimum conditions for the SPE of Cu(II) ions were investigated, and the method was subsequently applied to sea water, stream water, rice, tea, and tobacco samples for the determination of Cu(II) levels.

LC/ESI-MS method applied to characterization of flavonoids glycosides in B. forficata subsp. pruinosa

Bauhinia forficata is used in folk medicine for its hypoglycemiant effect. In the south of Brazil, the subspecies pruinosa is found in a region with the characteristic flora, pampa biome. This species has been consumed by the local population as a tea for diabetes treatment. We studied the chemical composition of hydroethanolic extracts using LC/ESI-MS. The leaf extracts were prepared by percolation with 50% (v/v) ethanol. The chromatographic analyses were performed using a reverse-phase system, gradient elution with acetonitrile:phosphoric acid 0.05%, and ESI-MS in the positive ion mode. The chemical profile of the flavonoids was suggested to involve four quercetin and kaempferol glycosides.

Avaliação da labilidade de alumínio em infusões de erva-mate empregando voltametria adsortiva de redissolução catódica

Infusions of yerba mate obtained at different stages of industrialization were evaluated to determine the bioavailable fraction of Al. Adsorptive Cathodic Stripping Voltammetry using DASA (complexing agent) was applied to determine the labile fraction of Al at pH 5.0 and pH 8.0 for the total fraction of dissolved Al. The results indicate that on average 60% of Al is complexed with organic compounds, minimizing their bioavailability; however, the labile fraction exceeds by up to 4 times the maximum weekly intake recommended by the World Health Organization.

USO DA RMN DE BAIXA RESOLUÇÃO NA AVALIAÇÃO DA DINÂMICA MOLECULAR DO Origanum vulgare

In this work, proton NMR relaxometry was used to measure the behavior of spin-lattice relaxation time with T1H as the time constant, and also of spin-spin relaxation time with the time constant T2H. These relaxometry parameters were determined to better understand the changes in the main structures present in commercial and in nature forms of origanum. The T1H relaxation data showed that the structures which had higher molecular mass were more sensitive to degradation with increased temperature treatment. According to the values of the T2H parameter, up to 150 degrees no significant change in the mobility and organization of water was observed. These data infer that the ideal cooking temperature and tea preparation mode for this herb should be around 100 degrees for the sample not to lose its characteristics. Also, it is not advisable to cook this herb at higher than 150 degrees but better to consume it at room temperature, especially give commercial herb has already been dehydrated.

ESTUDO DA RELAÇÃO ENTRE CONDIÇÕES DE POLPAÇÃO E PROPRIEDADES FÍSICO-MECÂNICAS DO PAPEL

RESUMOO objetivo deste estudo foi avaliar a influência da condição de polpação nas propriedades físico-mecânicas do papel branqueado de eucalipto. Para este estudo, foram utilizados cavacos de um híbrido clonal de eucalipto (Eucalyptus grandis x Eucalyptus urophylla) de 7 anos de idade. As polpas foram produzidas por duas condições distintas de polpação Lo-SolidsTM, sendo elas: Condição PI – álcali efetivo de 16,5%, sulfidez de 30% e temperatura de cozimento de 155 ºC; e Condição PII – álcali efetivo de 17,5%, sulfidez de 32% e temperatura de cozimento de 147 ºC. Ambos os cozimentos foram realizados buscando-se número kappa de 18,0 ± 0,5. Os branqueamentos foram realizados pela sequência de branqueamento OA(Ze)DP, buscando-se a alvura de 90,0 ± 1,0% ISO. Os refinos foram realizados a 0, 750, 1.500 e 3.000 níveis de revoluções e as correlações, feitas utilizando o índice de tração como variável independente da curva de refino. As propriedades físico-mecânicas do papel foram influenciadas pela condição de polpação, com aumento da drenabilidade, densidade aparente, índice TEA, alongamento e índice de rasgo para a polpa PI, considerando-se um mesmo índice de tração. A polpa PII, entretanto, atingiu o mesmo índice de tração da polpa PI com menor nível de refino e, consequentemente, menor gasto de energia. A condição de polpação PII foi mais adequada para a produção de papel por preservar mais a integridade das fibras, alcançando um mesmo índice de tração com um menor nível de refino.

Estudo endócrino reprodutivo e do comportamento sócio-sexual de sagui-de-tufo-preto (Callithrix penicillata) mantido em cativeiro

A comunicação do estado reprodutivo nos primatas da família Callithrichidae, depende principalmente dos comportamentos sócio-sexuais como um sistema de sinalização primário, uma vez que nestas espécies a ovulação não é percebida pelos machos. Neste trabalho, os padrões de comportamentos sócio-sexuais foram analisados em conjunto com as concentrações de metabólitos fecais dos esteróides sexuais progesterona (MFP), estradiol (MFE) e testosterona (MFT) em casais cativos de Sagüi-de-tufos-pretos (Callithrix penicillata), nas diferentes fases do ciclo ovariano. O grupo estudado era composto por quarto casais adultos, mantidos no Centro de Reabilitação de Animais Selvagens da prefeitura de São Paulo. Os padrões comportamentais foram registrados pelo método de amostragem focal por intervalo de tempo a cada 30 segundos, cinco vezes por semana, totalizando 14.400 registros por animal. A mensuração das concentrações de metabólitos fecais dos esteroides sexuais foram realizados pelo método de enzima imunoensaio (EIE). Os resultados obtidos dessas concentrações possibilitaram a determinação endócrina das fases do ciclo ovariano (folicular e luteal) e de suas respectivas durações, assim como a determinação da fase periovulatória. Foram caracterizados 31 ciclos ovarianos completos, com duração de 24,3±4,1 dias (média ±DP), sendo que a fase folicular compreendeu 13,04±4,8dias e a fase lútea 11,2±4,2 dias. Os comportamentos sócio-sexuais (marcação por cheiro, cheirar genitália, catação e apresentação sexual) e a variável "proximidade" mostraram-se significativamente mais prevalentes na fase periovulatória do que nas demais fases do ciclo. Não houve alteração das concentrações de MFT dos machos ao longo de todo o período estudado. A análise conjunta das concentrações de metabólitos fecais de esteróides sexuais e dos comportamentos sócio-sexuais possibilitou um melhor entendimento das relações endócrino-comportamentais e reprodutivas de C. penicillata.

Connexin hemichannels and cell-cell channels: comparison of properties

Connexin46 (Cx46) forms functional hemichannels in the absence of contact by an apposed hemichannel and we have used these hemichannels to study gating and permeation at the single channel level with high time resolution. Using both cell-attached and -excised patch configurations, we find that single Cx46 hemichannels exhibit some properties expected of half of a gap junction channel, as well as novel properties. Cx46 hemichannels have a large unitary conductance (~300 pS) and a relatively large pore as inferred from permeability to TEA. Both monovalent cations and anions can permeate, but cations are substantially more permeable. The open channel conductance shows marked inward rectification in symmetric salts. We find that the conductance and permeability properties of Cx46 cell-cell channels can be explained by the series addition of two hemichannels. These data suggest that the pore structures of unapposed hemichannels and cell-cell channels are conserved. Also like cell-cell channels, unapposed Cx46 hemichannels are closed by elevated levels of H+ or Ca2+ ions on the cytoplasmic face. Closure occurs in excised patches indicating that the actions of these agents do not require a soluble cytoplasmic factor. Fast (<0.5 ms) application of H+ to either side of the open hemichannel causes an immediate small reduction in unitary conductance followed by complete closure with latencies that are dependent on H+ concentration and side of application; sensitivity is much greater to H+ on the cytoplasmic side. Closure by cytoplasmic H+ does not require that the hemichannel be open. Thus, H+ ions readily permeate Cx46 hemichannels, but at high enough concentration close them by acting at a cytoplasmic site(s) that causes a conformational change resulting in complete closure. Extracellular H+ may permeate to act on the cytoplasmic site or act on a lower affinity extracellular site. Thus, the unapposed hemichannel is a valuable tool in addressing fundamental questions concerning the operation of gap junction channels that are difficult to answer by existing methods. The ability of Cx46, and perhaps other connexins, to form functional unapposed hemichannels that are opened by moderate depolarization may represent an unexplored role of connexins as mediators of transport across the plasma membrane.

The role of calcium, calcium-activated K+ channels, and tyrosine/kinase in psoralen-evoked responses in human melanoma cells

8-Methoxy psoralen (8-MOP) exerts a short-term (24 h) mitogenic action, and a long-term (48-72 h) anti-proliferative and melanogenic action on two human melanoma cell lines, SK-Mel 28 and C32TG. An increase of intracellular calcium concentration was observed by spectrofluorometry immediately after the addition of 0.1 mM 8-MOP to both cell lines, previously incubated with calcium probe fluo-3 AM (5 µM). The intracellular Ca2+ chelator BAPTA/AM (1 µM) blocked both early (mitogenic) and late (anti-proliferative and melanogenic) 8-MOP effects on both cell lines, thus revealing the importance of the calcium signal in both short- and long-term 8-MOP-evoked responses. Long-term biological assays with 5 and 10 mM tetraethylammonium chloride (TEA, an inhibitor of Ca2+-dependent K+ channels) did not affect the responses to psoralen; however, in 24-h assays 10 mM TEA blocked the proliferative peak, indicating a modulation of Ca2+-dependent K+ channels by 8-MOP. No alteration of cAMP basal levels or forskolin-stimulated cAMP levels was promoted by 8-MOP in SK-Mel 28 cells, as determined by radioimmunoassay. However, in C32TG cells forskolin-stimulated cAMP levels were further increased in the presence of 8-MOP. In addition, assays with 1 µM protein kinase C and calcium/calmodulin-dependent kinase inhibitors, Ro 31-8220 and KN-93, respectively, excluded the participation of these kinases in the responses evoked by 8-MOP. Western blot with antibodies anti-phosphotyrosine indicated a 92% increase of the phosphorylated state of a 43-kDa band, suggesting that the phosphorylation of this protein is a component of the cascade that leads to the increase of tyrosinase activity.

Participation of ATP-sensitive K+ channels in the peripheral antinociceptive effect of fentanyl in rats

We examined the effect of several K+ channel blockers such as glibenclamide, tolbutamide, charybdotoxin (ChTX), apamin, tetraethylammonium chloride (TEA), 4-aminopyridine (4-AP), and cesium on the ability of fentanyl, a clinically used selective µ-opioid receptor agonist, to promote peripheral antinociception. Antinociception was measured by the paw pressure test in male Wistar rats weighing 180-250 g (N = 5 animals per group). Carrageenan (250 µg/paw) decreased the threshold of responsiveness to noxious pressure (delta = 188.1 ± 5.3 g). This mechanical hyperalgesia was reduced by fentanyl (0.5, 1.5 and 3 µg/paw) in a peripherally mediated and dose-dependent fashion (17.3, 45.3 and 62.6%, respectively). The selective blockers of ATP-sensitive K+ channels glibenclamide (40, 80 and 160 µg/paw) and tolbutamide (80, 160 and 240 µg/paw) dose dependently antagonized the antinociception induced by fentanyl (1.5 µg/paw). In contrast, the effect of fentanyl was unaffected by the large conductance Ca2+-activated K+ channel blocker ChTX (2 µg/paw), the small conductance Ca2+-activated K+ channel blocker apamin (10 µg/paw), or the non-specific K+ channel blocker TEA (150 µg/paw), 4-AP (50 µg/paw), and cesium (250 µg/paw). These results extend previously reported data on the peripheral analgesic effect of morphine and fentanyl, suggesting for the first time that the peripheral µ-opioid receptor-mediated antinociceptive effect of fentanyl depends on activation of ATP-sensitive, but not other, K+ channels.

The methyl ester of rosuvastatin elicited an endothelium-independent and 3-hydroxy-3-methylglutaryl coenzyme A reductase-independent relaxant effect in rat aorta

The relaxant effect of the methyl ester of rosuvastatin was evaluated on aortic rings from male Wistar rats (250-300 g, 6 rats for each experimental group) with and without endothelium precontracted with 1.0 µM phenylephrine. The methyl ester presented a slightly greater potency than rosuvastatin in relaxing aortic rings, with log IC50 values of -6.88 and -6.07 M, respectively. Unlike rosuvastatin, the effect of its methyl ester was endothelium-independent. Pretreatment with 10 µM indomethacin did not inhibit, and pretreatment with 1 mM mevalonate only modestly inhibited the relaxant effect of the methyl ester. Nω-nitro-L-arginine methyl ester (L-NAME, 10 µM), the selective nitric oxide-2 (NO-2) inhibitor 1400 W (10 µM), tetraethylammonium (TEA, 10 mM), and cycloheximide (10 µM) partially inhibited the relaxant effect of the methyl ester on endothelium-denuded aortic rings. However, the combination of TEA plus either L-NAME or cycloheximide completely inhibited the relaxant effect. Inducible NO synthase (NOS-2) was only present in endothelium-denuded aortic rings, as demonstrated by immunoblot with methyl ester-treated rings. In conclusion, whereas rosuvastatin was associated with a relaxant effect dependent on endothelium and hydroxymethylglutaryl coenzyme A reductase in rat aorta, the methyl ester of rosuvastatin exhibited an endothelium-independent and only slightly hydroxymethylglutaryl coenzyme A reductase-dependent relaxant effect. Both NO produced by NOS-2 and K+ channels are involved in the relaxant effect of the methyl ester of rosuvastatin.