Caracterização e quantificação de marcadores químicos do extrato hidroetanólico das folhas de Kalanchoe brasiliensis Cambess

Kalanchoe brasiliensis Cambess (Crassulaceae), commonly known as saião , coirama branca , folha grossa , is originally from Brazil and commonly found in São Paulo to Bahia, mainly in the coastal zone. Regarding of biological activities, most preclinical studies were found in the literature, mainly about the anti-inflammatory activity of extracts obtained from leaves and / or aerial parts of K. brasiliensis. As regards the chemical constitution, it has been reported mainly the presence of flavonoids in the leaves of the species, but until this moment did not knows which are the active compounds. Although it is a species widely used in traditional medicine in Brazil, there is no monograph about the quality parameters of the plant drug. In this context, this study aims to characterize and quantify the chemical markers of hydroethanolic extract (HE) from the leaves of K. brasiliensis, which can be used in quality control of plant drug and derivatives obtained from this species. The methodology was divided into two parts: i. Phytochemical study: to fractionate, isolate and characterizate of the chemical (s) marker (s) of the HE from the leaves of K. brasiliensis; ii. To Developed validate of analytical method by High Performance Liquid Chromatography (HPLC)-diode array detector (DAD) to quantify the chemical (s) marker (s) of the EH. i. The EH 50% was prepared by turbo extraction method. It was then submitted to liquid-liquid partition, obtaining dichloromethane, n-butanol and ethyl acetate (AcOEt) fractions. The AcOEt fraction was selected to continue the fractionation process, because it has a chemical profile rich in flavonoids. The acOEt fraction was submitted to column chromatography using different systems for obtaining the compound Kb1. To identify this compound, it was submitted to UV analysis ii. For quantitative analysis, the EH was analyzed by HPLC, using different methods. After selecting the most appropriate method, which showed satisfactory resolution and symmetrical peaks, it was validated according to parameters in the RE 899/2003. As result, it was obtained from the AcOEt fraction the compound Kb1 (2.7 mg). Until this moment, the basic nucleus was characterized by UV analysis using shift reagents. The partial chemical structure of the compound Kb1 was identified as a flavonol, containing hydroxyls in 3 , 4 position (ring A), 5 and 7 free (ring B) and a replacement of the C3 hydroxyl by a sugar. As the analysis were performed in the HPLC coupled to a DAD, we observed that the UV spectrum of the major peaks of EH from K. brasiliensis shown similar UV spectrum. According to the literature, it has been reported the presence of patuletin glycosydes derivatives in the leaves of this species. Therefore, it is suggested that the compound Kb1 is glycosylated patuletin derivative. Probably the sugar (s) unit(s) are linked in the C3 in the C ring. . Regarding the development of HPLC analytical method, the system used consists of phase A: water: formic acid (99,7:0,3, v / v) and phase B: methanol: formic acid (99,7:0,3, v / v), elution gradient of 40% B - 58% B in 50 minutes, ccolumn (Hichrom ®) C18 (250x4, 0 mm, 5 μm), flow rate 0.8 mL / min, UV detection at 370 nm, temperature 25 ° C. In the analysis performed with the co-injection of thecompound Kb1 + HE of K. brasiliensis was observed that it is one of the major compounds with a retention time of 12.47 minutes and had a content of 15.3% in EH of leaves from K. brasiliensis. The method proved to be linear, precise, accurate and reproducible. According to these results, it was observed that compound Kb1 can be used as a chemical marker of EH from leaves of K. brasiliensis, to assist in quality control of drug plant and its derivatives

Balanço energético comparativo para rotas de escoamento de soja

O presente trabalho objetivou analisar o fluxo energético no escoamento de soja da região Centro-Oeste do Brasil, considerando as rotas atualmente existentes, mais precisamente, do município de Rio Verde -GO até o porto de Santos - SP, visto que este é um grande polo exportador de granéis sólidos. Foram selecionadas duas rotas para a análise, uma contemplando a multimodalidade, ou seja, os modais rodoviário, ferroviário e hidroviário, e outra, uma única modalidade, o modal rodoviário. A conversão dos fatores físicos e operacionais em unidades energéticas foi realizada por meio de coeficientes energéticos levantados junto à literatura. Os resultados obtidos indicaram, pela rota 1 multimodal, um gasto energético específico maior para o modal rodoviário (0,50 MJ km-1 t-1), seguido pelo modal ferroviário (0,42 MJ km-1 t-1) e, em terceiro, o modal hidroviário (0,22 MJ km-1 t-1). Pela rota 2, unimodal rodoviário, o resultado indicou 0,50 MJ km-1 t-1. Nas participações de energia apresentadas, comparando as rotas 1 e 2, a que apresentou maior gasto energético específico total foi a rota 2 (0,50 MJ km-1 t-1), seguida pela rota 1 (0,34 MJ km-1 t-1).

Recuperação e purificação de quitosanases usando adsorção em leito expandido com streamline DEAE com modelagem e simulação usando redes neurais

Expanded Bed Adsorption (EBA) is an integrative process that combines concepts of chromatography and fluidization of solids. The many parameters involved and their synergistic effects complicate the optimization of the process. Fortunately, some mathematical tools have been developed in order to guide the investigation of the EBA system. In this work the application of experimental design, phenomenological modeling and artificial neural networks (ANN) in understanding chitosanases adsorption on ion exchange resin Streamline® DEAE have been investigated. The strain Paenibacillus ehimensis NRRL B-23118 was used for chitosanase production. EBA experiments were carried out using a column of 2.6 cm inner diameter with 30.0 cm in height that was coupled to a peristaltic pump. At the bottom of the column there was a distributor of glass beads having a height of 3.0 cm. Assays for residence time distribution (RTD) revelead a high degree of mixing, however, the Richardson-Zaki coefficients showed that the column was on the threshold of stability. Isotherm models fitted the adsorption equilibrium data in the presence of lyotropic salts. The results of experiment design indicated that the ionic strength and superficial velocity are important to the recovery and purity of chitosanases. The molecular mass of the two chitosanases were approximately 23 kDa and 52 kDa as estimated by SDS-PAGE. The phenomenological modeling was aimed to describe the operations in batch and column chromatography. The simulations were performed in Microsoft Visual Studio. The kinetic rate constant model set to kinetic curves efficiently under conditions of initial enzyme activity 0.232, 0.142 e 0.079 UA/mL. The simulated breakthrough curves showed some differences with experimental data, especially regarding the slope. Sensitivity tests of the model on the surface velocity, axial dispersion and initial concentration showed agreement with the literature. The neural network was constructed in MATLAB and Neural Network Toolbox. The cross-validation was used to improve the ability of generalization. The parameters of ANN were improved to obtain the settings 6-6 (enzyme activity) and 9-6 (total protein), as well as tansig transfer function and Levenberg-Marquardt training algorithm. The neural Carlos Eduardo de Araújo Padilha dezembro/2013 9 networks simulations, including all the steps of cycle, showed good agreement with experimental data, with a correlation coefficient of approximately 0.974. The effects of input variables on profiles of the stages of loading, washing and elution were consistent with the literature

Recuperação e purificação de proteínas do soro de queijo tipo coalho usando cromatografia de troca iônica e interação hidrofóbica em leito na forma expandida

Expanded Bed Adsorption plays an important role in the downstream processing mainly for reducing costs as well as steps besides could handling cells homogenates or fermentation broth. In this work Expanded Bed Adsorption was used to recover and purify whey proteins from coalho cheese manufacture using Streamline DEAE and Streamline SP both ionic resins as well as a hydrophobic resin Streamline Phenyl. A column of 2.6 cm inner diameter with 30 cm in height was coupled to a peristaltic pump. Hydrodynamics study was carried out with the three resins using Tris-HCl buffer in concentration of 30, 50 and 70 mM, with pH ranging from 7.0 to 8.0. In this case, assays of the expansion degree as well as Residence Time Distribution (RTD) were carried out. For the recovery and purification steps, a whey sample of 200 mL, was submitted to a column with 25mL of resin previously equilibrated with Tris/HCl (50 mM, pH 7.0) using a expanded bed. After washing, elution was carried out according the technique used. For ionic adsorption elution was carried out using 100 mL of Tris/HCl (50 mM, pH 7.0 in 1M NaCl). For Hydrophobyc interaction elution was carried out using Tris/HCl (50 mM, pH 7.0). Adsorption runs were carried out using the three resins as well as theirs combination. Results showed that for hydrodynamics studies a linear fit was observed for the three resins with a correlation coefficient (R2) about 0.9. In this case, Streamline Phenyl showed highest expansion degree reaching an expansion degree (H0/H) of 2.2. Bed porosity was of 0.7 when both resins Streamline DEAE and Streamline SP were used with StremLine Phenyl showing the highest bed porosity about 0.75. The number of theorical plates were 109, 41.5 and 17.8 and the axial dipersion coefficient (Daxial) were 0.5, 1.4 and 3.7 x 10-6 m2/s, for Streamline DEAE, Streamline SP and Streamline Phenyl, respectively. Whey proteins were adsorved fastly for the three resins with equilibrium reached in 10 minutes. Breakthrough curves showed that most of proteins stays in flowthrough as well as washing steps with 84, 77 and 96%, for Streamline DEAE, Streamline SP and Streamline Phenyl, respectively. It was observed protein peaks during elution for the three resins used. According to these peaks were identified 6 protein bands that could probably be albumin (69 KDa), lactoferrin (76 KDa), lactoperoxidase (89 KDa), β-lactoglobulin (18,3 KDa) e α-lactoalbumin (14 KDa), as well as the dimer of beta-lactoglobulin. The combined system compound for the elution of Streamline DEAE applied to the Streamline SP showed the best purification of whey proteins, mainly of the α-lactoalbumina

A Transposição Didádica dos Gêneros do domínio do discursivo publicitários nos livros didáticos

Advertising text has been a subject of many investigations, because of its multimodal universe. Embodied by a linguistic and discursive materiality laying on scene persuasion, argumentation and power imagery of multissemiotics elements, the advertising text acts as an instrument of power, creating and destroying, promising and denying (CARVALHO, 2007). Advertising not only invites us to act for it, but directs us to look at it. It was under this moving look - of charm and interrogations - that discussions raised in this research were born. Investigations are directed to the school environment, in special, for the discursive advertising domain in Portuguese Language Textbook. It is from this environment that was born our research whose main objective is to analyze how does the didactic transposition of textual genres, described by Marcuschi (2008) as belonging to the "advertising" discourse domain (focusing on advertising genre) in didactic books teaching Portuguese Language before and after the advent of the PCN. Textbooks taken as reference for the study are historically situated in the 90s of the twentieth century and 10 century. Such books refer to elected 7th and 8th grades, currently corresponding to the 8th and 9th grades of elementary school. The choice is justified by the fact that in these Textbook teaching series the presence of advertising domain is recurrent and "didactized". In addition, we are also concerned in analyzing books that circulated around us and our regional reality. Hence, we elected books that were used by two public high schools: Municipal School Clementina Ana da Conceição in Jaçanã city in the Rio Grande do Norte state, State School for Elementary and High school Carlota Barreira in Areia city, Paraíba state. In our research the following categories of analysis were highlighted: (1) presence of advertising in DB, (2) fluctuation terminology: concepts and classifications; (3) The complexity of concepts facilitation, (4) what they propagate, and from which nature are the explored advertisements. From our analysis, we observe how the treatment of textual "advertising" genres have been inserted into Textbooks, and how occurs, in general, their didactic transposition. Focusing on the issue of fluctuating terminology, we noted the difficulty in drawing boundaries between the genres of advertising domain in the Textbooks. However, this also would result in the complexity - in the field of scientific knowledge - of delimiting genres of the same domain. To accomplish our studies, it was required a thorough and systematic dialogue with theories regarding the concept of "Didactic Transposition", due to the theoretical Chevallard Yves (1991), research on the textual genres - Bezerra (2005), Marcuschi (2008), Bazerman (2005), Swales (2004), among others - and studies involving the field of "Advertising" - Sandmann (2002), Carvalho (2007) and others

Letramento no trânsito: eventos e práticas na formação de condutores de veículos

In the contemporary society, the language is presented in all social spaces and assumes many different purposes in order to meet the needs that emerge from each of these sphere. In traffic, this reality is not different. To guide vehicles, it is necessary to know, by means of reading artifacts, what the legislation establishes in what concerns the way to act in this domain. Thus, this works aims at describing the practices of literacy held in events of driver trainings and know the expectations generated by drivers/learners from this training. In theoretical terms, it anchors in Literacy Studies, comprehended here as social practices (BARTON; HAMILTON, 1998; KLEIMAN, 1995, 2008; MORTATTI, 2004; STREET, 1984; OLIVEIRA, 2008, 2010; ROJO, 2009; PAZ, 2008). Genre Theory (BRONCKART, 2004, 1999; OLIVEIRA, 2010) and in your multimodal instance (KRESS; VAN LEEUWEN, 1996; DIONÍSIO, 2006). In terms of methodology, it follows the bias of qualitative research, because of its ethnographic nature (BOGDAN; BIKLEN, 1994; MINAYO, 2010; CANÇADO, 1994; CHIZZOTTI, 2005). The research corpus was generated by reading the Brazilian Traffic Code, by observing the literacy events held in Drivers Training Centers of Natal, analysis of course books used in these events, plus questionnaires with open and closed questions and semistructured interviews. The collaborators are constituted of drivers in training, and instructors who work in this field. The analyses show significant contributions regarding the placement more committed of future drivers with the welfare and safety of those who use the public roads, from the practice or reading done during the traffic training. The contribution of this work lies in the possibility to expand the discussion about the language practice uses regarding the training for the traffic, more specifically, the training of drivers of vehicles

O desenvolvimento da interlíngua na aprendizagem da escrita em inglês em uma escola bilíngue: um estudo exploratório

The aim of this study is to investigate the development of written Interlanguage in English as an Additional Language (AL) by students in the 2nd grade of Ensino Fundamental I in a bilingual school in the city of Natal-RN. For this purpose two research questions guided this study: (a) which hypotheses could be inferred from the writing development of the bilingual learners of English as AL? and, (b) what is the impact of the type of input monomodal or multimodal in the Interlanguage development in the AL of bilingual learners? The 38 learners were divided into a control group, with 21 learners exposed to monomodal input, and an experimental group, with 17 learners exposed to multimodal input, and pre and post-tests were applied to both groups. A mixed methods research design was conducted (DÖRNYEI, 2007) to involve both qualitative and quantitative data collection and analysis. The qualitative aspect comprehended descriptive characteristics that interpreted the central cognitive processes in the acquisition of writing in AL by the learners. Through these interpretations, it was possible to understand the constitution of written Interlanguage (SELINKER, 1972) according to the data generated by the learners. The quantitative data were presented as the results generated from the experimental design. Thus, they narrowed the relations between the dependent variable the writing development, that is, how close it is to the target form which was modified throughout the process by the independent variable the quality of input (VAN PATTEN, 2002, GASS, 1997, SCHMIDT, 1986, PARADIS, 2009; 2010, ELLIS, 1995), which, being monomodal or multimodal, its function was possibly to alter the route of acquisition. The quantitative results pointed towards significant gains by the experimental group, which had multimodality present, suggesting that the learners in this group seem to have been more able to cognitively register (SCHIMDT, 1990) aspects of learning than the learners in the control group

O ensaio jornalístico na revista Veja: uma análise multidimensional

This work has as objective generality to make a multidimensional analysis in the genre journalistic assay, communicative genre that, beyond complex and multimodal, presents hybrid characteristics. Specifically, with the intention to propose defining criteria of the cited genre, this research looks for to establish differences and similarities between the assay and other genres of the same sphere, from the description and interpretation of used multimodal resources. The analysis of the formal, schematical and rhetorical resources identified in the formatting of the journalistic assay sample that the analyses are supported in the socio-semiotic and socio-rhetorical approaches. In the formal dimension, we contemplate elements that constitute design of the text, including the forms of representation from the typography, the colors, images, as well as the aspects communicative-linguistics: the modalization indices, the communicative operators and the category time; in the schematical dimension, we present the organizational structure, considering the rhetorical movements postulates for Swales (1990) and in the rhetorical dimension we observe the categories: who writes, for who it writes, on what it writes and where writes. The adopted methodologicals postulates are of qualitative nature and the procedure is documentary, data that in we are valid them written texts of this genre as analysis object. Corpus it is constituted by a composed sample for 14 extracted texts of a set of 173 propagated journalistic assays weekly for the magazine Veja, in the period between August of 2004 and January of 2008. The analysis of the data showed that the journalistic assay, object of this study, materializes through multiple symbolic representations and multiple subjects that turn since a small episode of the daily facts of great social relevance in the present time, of historical and cultural nature, nationwide or international. Used for the first time by Montaigne in 1580, to assign, in saying of the proper author, written fast on its life and historical events, which could nor be remembered later , the term `assay' was enriched with other specifications, of form to enclose the one that if they call scientific assay today, academic assay, journalistic assay and other types of specific assays. These denominations have to see with the enrollment of the members of diverse of practices communities, in virtue of the multiplicity of activities carried through in these spheres. The conclusions the one that we arrive had been the following ones: 1. the discursive genre is not a pure entity, in virtue of the multiplicity of situations where the sorts if insert in the social actions; 2. the institutions define the configuration of one definitive genre, also its proper assignment, since for backwards of all discursive genre a voice exists to discipline - institutional voice, and in the case of the assays for analyzed us, the institutional voice if it presents, really, as a defining trace; 3. the journalistic assay, for its multiple symbolic representations, multiple subjects and for passing explicit or implicit opinions of its author, resembles it other genres, being able, therefore, to be inserted in a colony of opinionatives genres

Application of an HPLC method for analysis of propolis extract

Propolis obtained from honeybee hives has been widely used in medicine, cosmetics, and industry due to its versatile biological activities (antioxidant, antimicrobial, fungicidal, antiviral, antiulcer, immunostimulating, and cytostatic). These activities are mainly attributed to the presence of flavonoids in propolis, which points out the interest in quantifying these constituents in propolis preparations, as well as validation of analytical methodologies. High-performance liquid chromatography (HPLC) methods have been reported to quantify isolated flavonoids or these compounds in complex biological matrices, such as herbal raw materials and extractive preparations. An efficient, precise, and reliable method was developed for quantification of propolis extractive solution using HPLC with UV detection. The chromatograms were obtained from various gradient elution systems (GES) tested in order to establish the ideal conditions for the analysis of propolis extractive solution, using methanol and water: acetonitrile (97.5 : 2.5, v/v) as mobile phase. Gradient reversed phase chromatography was performed using a stainless steel column (250 x 4.6 mm i.d., 5 mum) filled with Chromsep RP 18 (Varian), column temperature at 30.0 +/- 0.1degreesC and detection at 310 nm. The main validation parameters of the method were also determined. The method showed linearity for chrysin in the range 0.24-2.4 mug mL(-1) with good correlation coefficients (0.9975). Precision and accuracy were determined. The obtained results demonstrate the efficiency of the proposed method. The analytical procedure is reliable and offers advantages in terms of speed and cost of reagents.

Major carotenoid composition of Brazilian Valencia orange juice: Identification and quantification by HPLC

The carotenoid composition of Brazilian Valencia orange juice was determined by open column chromatography (OCC) and high-performance liquid chromatography. Carotenoid pigments were extracted using acetone and saponified using 10% methanolic potassium hydroxide. Sixteen pigments were isolated by OCC and identified as alpha-carotene, zeta-carotene, beta-carotene, alpha-cryptoxanthin, beta-cryptoxanthin, lutein-5,6-epoxide, violaxanthin, lutein, antheraxanthin, zeaxanthin, luteoxanthin A, luteoxanthin B, mutatoxanthin A, mutatoxanthin B, auroxanthin B and trollichrome B. Thirteen carotenoid pigments were separated using a ternary gradient (acetonitrile-methanol-ethyl acetate) elution on a C-18 reversed-phase column. Among these, violaxanthin, lutein, zeaxanthin, beta-cryptoxanthin, zeta-carotene, alpha-carotene, and beta-carotene were quantified. The total carotenoid content was 12 +/- 6.7 mg/1, and the major carotenoids were lutein (23%), beta-cryptoxanthin (21%), and zeaxanthin (20%). 2005 Elsevier Ltd. All rights reserved.

Development and validation of an analytical method by RP-HPLC for quantification of sibutramine hydrochloride in pharmaceutical capsules

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caracterização e hidrólise in vitro da globulina principal de grão-de-bico (Cicer arietinum L.), var. IAC-Marrocos

No presente estudo procedeu-se ao isolamento e caracterização da fração globulina majoritária (11 S) de grão-de-bico, var. IAC-Marrocos. A globulina majoritária extraída foi isolada por cromatografia de filtração em gel e de troca-iônica mostrando apenas uma banda de proteína na eletroforese em gel de poliacrilamida. A globulina majoritária, após passagem em coluna de Sephadex, revelou duas bandas protéicas de 55 e 52,5kDa e três bandas menores em gel de poliacrilamida dodecilsulfato de sódio. Na presença de 2-mercaptoetanol 6 polipeptídios na faixa de 18 a 42kDa foram revelados na eletroforese. A globulina isolada foi submetida à ação da tripsina e quimotripsina onde a forma nativa mostrou-se resistente à ação enzimática enquanto o aquecimento (96 e 121°C/15min) não foi suficiente para aumentar a susceptibilidade à hidrólise, significativamente. Adição de NaCl 0,3M levou a um aumento da estabilidade estrutural com menor susceptibilidade à digestão proteolítica, fato em parte perdido com o aquecimento. As hidrólises foram acompanhadas por eletroforese em gel de poliacrilamida dodecilsulfato de sódio.

Effect of chickpea (Cicer arietinum L.) germination on the major globulin content and in vitro digestibility

A germinação das sementes de grão-de-bico foi acompanhada por um período de 6 dias, no qual pequenas variações nos teores de nitrogênio e globulina total foram registradas. A globulina majoritária (tipo 11 S) apresentou maiores variações após o quarto dia de germinação. A natureza e distribuição da fração globulina majoritária isolada na cromatografia em Sepharose CL-6B mostrou pequenas modificações ao final do período de germinação. A eletroforese em gel de poliacrilamida com dodecilssulfato de sódio do pico eluído na cromatografia em Sepharose CL-6B demonstra modificações nas bandas de proteínas entre os pesos moleculares de 20 e 30 kDa e acima de 60 kDa, indicando degradação protéica durante o período. Atividade proteolítica foi detectada na fração albumina da semente que aumentou até o quarto dia, seguido de queda até o sexto dia de germinação, quando da utilização de globulina total isolada da semente e caseína como substratos. Farinha de grão-de-bico, frações albumina e globulina total isoladas não apresentaram aumento na digestibilidade in vitro; entretanto, a fração globulina majoritária isolada foi mais suscetível à hidrólise após germinação.

Semipreparative enantioseparation of methamidophos by HPLC-UV and preliminary in vitro study of butyrylcholinesterase inhibition

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reactive power dispatch and planning using a non-linear branch-and-bound algorithm

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)