Extraction and purification of triterpenic acids from eucalypt bark

A indústria da pasta e do papel é um sector importante da economia mundial, particularmente a que assenta em espécies de Eucalyptus. No entanto, essas indústrias geram quantidades significativas de correntes secundárias de subprodutos e resíduos de biomassa que podem ser exploradas em aplicações de valor acrescentado em vez de serem queimadas para produção de energia. Um exemplo nobre pode ser a produção de ácidos triterpénicos com estruturas dos tipos lupano, oleanano e ursano, dada a sua abundância em alguns destes resíduos, em particular na casca, adotando o conceito de biorrefinaria integrada numa fábrica de pasta. Estes compostos são conhecidos pelas suas inúmeras actividades biológicas, por exemplo, antioxidante, anti-inflamatória e antitumoral, e podem encontrar aplicações em produtos de elevado valor, tais como cosméticos, nutracêuticos ou farmacêuticos. Neste sentido, o estudo das potencialidades das cascas das espécies de eucalipto mais exploradas enquanto fontes de compostos triterpénicos é um tópico relevante. Por conseguinte, foram analisados e comparados em pormenor os teores e composições em ácidos triterpénicos (TTAs) das cascas externas de várias espécies de eucalipto (E. globulus, E. grandis, E. urograndis, E. maidenii e E. nitens). Os teores dos principais TTAs identificados nestas espécies variaram entre 4.5 g/kg no E. urograndis e 21.6 g/kg no E. nitens. Observou-se que as cascas externas de Eucalyptus de zonas temperadas e Mediterrânicas, nomeadamente E. nitens e E. globulus, são mais ricas em TTAs que as espécies de regiões tropicais e subtropicais. Além disso, a casca externa do E. globulus é claramente a mais rica em ácidos com estruturas do tipo ursano enquanto a do E. nitens é a mais rica em ácidos do tipo oleanano e lupano. Estes resultados levaram-nos a estudar a extração dos TTAs da casca de Eucalyptus, bem como a sua posterior concentração e purificação, a qual foi efetuada por extração sólido-líquido convencional combinada com a precipitação de solutos, e por extração com fluidos supercríticos (SFE). No que diz respeito à primeira abordagem referida, foi desenvolvido neste trabalho um método patenteado que permite obter extratos enriquecidos em TTAs das cascas de eucalipto baseado em tecnologias disponíveis no imediato. Em relação à segunda abordagem, e de forma a apostar em processos de baixo impacto ambiental exigidos pelas biorrefinarias do futuro, a SFE surge como uma opção natural. Assim, foi efetuada a SFE da casca caduca do E. globulus com dióxido de carbono puro e modificado para recuperar a fração de TTAs, e os resultados foram comparados com os obtidos por extração em Soxhlet com diclorometano. Foram realizados estudos preliminares sobre a influência da pressão (100-200 bar), a adição de co-solvente (0, 5 e 8% m/m de etanol), e operação em múltiplos passos a fim de avaliar a aplicabilidade da alternativa supercrítica para a sua produção eficiente e selectiva. Os resultados destacaram a influência da pressão e o importante papel resumo (cont.) desempenhado pelo co-solvente neste processo, cujo efeito foi mais relevante do que o aumento da pressão em várias dezenas de bar. Este trabalho foi depois otimizado, usando o planeamento factorial de experiências e a metodologia de superfícies de resposta, para analisar a influência da temperatura (40-60 ºC), pressão (100-200 bar), e teor de etanol (0.0-5.0% m/m) na recuperação dos TTAs e respectiva concentração nos extractos. Nestes intervalos, as melhores condições de operação encontradas foram 200 bar, 40 °C e 5% de etanol, para as quais os modelos de regressão estatisticamente validados previram um rendimento de extração de 1.2% com 50% de concentração em TTAs, correspondendo ao rendimento em TTAs de 5.1 g/kg de casca e uma recuperação de 79.2% comparativamente ao valor do Soxhlet. Os TTAs livres e acetilados apresentaram tendências de extracção bastante distintas devido às suas diferentes afinidades para o CO2 causadas pelas diferentes polaridades: os derivados acetilados aproximam-se de um patamar máximo a cerca de 200 bar e 5% de etanol, enquanto a extração dos TTAs livres apresenta uma tendência sempre crescente no intervalo de condições estudado. Foram também medidas curvas cumulativas de SFE da casca do E. globulus de forma a analisar o comportamento cinético do processo em termos de rendimento total, rendimento em TTAs, rendimento em TTAs livres, rendimento em TTAs acetilados, e concentração dos TTAs nos extractos. Foi analisada a influência da pressão, temperatura, teor de co-solvente e caudal do dióxido de carbono sobre as respostas anteriores. Os dados experimentais foram modelados com os modelos Logístico, de Dessorção, de Placa Plana Simples, e de Difusão. Na globalidade, os resultados confirmaram que a pressão e o teor de etanol têm um efeito significativo sobre as curvas de extração, os rendimentos finais e as concentrações dos extratos, e mostraram a presença de limitações externas à transferência de massa em alguns ensaios. Mais uma vez, as famílias individuais de TTAs livres e acetilados apresentaram diferentes tendências de extracção. A modelação permitiu-nos confirmar não só o importante papel desempenhado pela difusão intraparticular na SFE, mas também a contribuição da resistência no filme em alguns ensaios. Após a análise de todos os resultados, foi efetuado um ensaio em duas etapas em série, possibilitando o enriquecimento do teor em TTAs no extracto devido às diferentes condições adotadas em cada etapa. Por último, um éster metílico de um ácido triterpénico do tipo oleanano - morolato de metilo - foi identificado pela primeira vez enquanto componente da casca de Eucalyptus na casca externa do Eucalyptus grandis x globulus, onde ocorre em teores elevados. A sua extração com CO2 supercrítico foi também realizada, visando a conceção de uma alternativa de extração ambientalmente benigna para este composto. A 200 bar e 60 ºC, a remoção do morolato de metilo atingiu um patamar às 6 h para 5.1 kg h-1 de CO2 / kg de casca. Em geral, e de forma semelhante à SFE da casca do E. globulus, os TTAs acetilados foram mais significativamente extraídos quando comparados com os seus ácidos livres, o que está diretamente relacionado com a natureza menos polar destas moléculas. O trabalho apresentado nesta tese é uma contribuição para a valorização de uma corrente de biomassa com baixo valor na indústria de pasta em duas vertentes complementares. Por um lado, aumentou o conhecimento da composição lipofílica das cascas de Eucalyptus spp. com interesse comercial para a produção de pasta, destacando o seu potencial enquanto fontes de ácidos triterpénicos. Por outro lado, foram desenvolvidos dois processos alternativos e facilmente integráveis numa fábrica de pasta para a sua exploração a partir da casca: um baseado em tecnologias convencionais bem estabelecidas a nível industrial, prevendo a sua aplicação a curto prazo, e um outro baseado na SFE, seguindo as tendências das futuras biorrefinarias.

Mathematical modelling of supercritical CO2 extraction of volatile oils from aromatic plants

The modelling of the experimental data of the extraction of the volatile oil from six aromatic plants (coriander, fennel, savoury, winter savoury, cotton lavender and thyme) was performed using five mathematical models, based on differential mass balances. In all cases the extraction was internal diffusion controlled and the internal mass transfer coefficienty (k(s)) have been found to change with pressure, temperature and particle size. For fennel, savoury and cotton lavender, the external mass transfer and the equilibrium phase also influenced the second extraction period, since k(s) changed with the tested flow rates. In general, the axial dispersion coefficient could be neglected for the conditions studied, since Peclet numbers were high. On the other hand, the solute-matrix interaction had to be considered in order to ensure a satisfactory description of the experimental data.

Compositional evolution with mass transfer in closed systems

Evolution of compositions in time, space, temperature or other covariates is frequent in practice. For instance, the radioactive decomposition of a sample changes its composition with time. Some of the involved isotopes decompose into other isotopes of the sample, thus producing a transfer of mass from some components to other ones, but preserving the total mass present in the system. This evolution is traditionally modelled as a system of ordinary di erential equations of the mass of each component. However, this kind of evolution can be decomposed into a compositional change, expressed in terms of simplicial derivatives, and a mass evolution (constant in this example). A rst result is that the simplicial system of di erential equations is non-linear, despite of some subcompositions behaving linearly. The goal is to study the characteristics of such simplicial systems of di erential equa- tions such as linearity and stability. This is performed extracting the compositional dif ferential equations from the mass equations. Then, simplicial derivatives are expressed in coordinates of the simplex, thus reducing the problem to the standard theory of systems of di erential equations, including stability. The characterisation of stability of these non-linear systems relays on the linearisation of the system of di erential equations at the stationary point, if any. The eigenvelues of the linearised matrix and the associated behaviour of the orbits are the main tools. For a three component system, these orbits can be plotted both in coordinates of the simplex or in a ternary diagram. A characterisation of processes with transfer of mass in closed systems in terms of stability is thus concluded. Two examples are presented for illustration, one of them is a radioactive decay

Deficits in agency in schizophrenia, and additional deficits in body image, body schema, and internal timing, in passivity symptoms

Individuals with schizophrenia, particularly those with passivity symptoms, may not feel in control of their actions, believing them to be controlled by external agents. Cognitive operations that contribute to these symptoms may include abnormal processing in agency as well as body representations that deal with body schema and body image. However, these operations in schizophrenia are not fully understood, and the questions of general versus specific deficits in individuals with different symptom profiles remain unanswered. Using the projected-hand illusion (a digital video version of the rubber-hand illusion) with synchronous and asynchronous stroking (500 ms delay), and a hand laterality judgment task, we assessed sense of agency, body image, and body schema in 53 people with clinically stable schizophrenia (with a current, past, and no history of passivity symptoms) and 48 healthy controls. The results revealed a stable trait in schizophrenia with no difference between clinical subgroups (sense of agency) and some quantitative (specific) differences depending on the passivity symptom profile (body image and body schema). Specifically, a reduced sense of self-agency was a common feature of all clinical subgroups. However, subgroup comparisons showed that individuals with passivity symptoms (both current and past) had significantly greater deficits on tasks assessing body image and body schema, relative to the other groups. In addition, patients with current passivity symptoms failed to demonstrate the normal reduction in body illusion typically seen with a 500 ms delay in visual feedback (asynchronous condition), suggesting internal timing problems. Altogether, the results underscore self-abnormalities in schizophrenia, provide evidence for both trait abnormalities and state changes specific to passivity symptoms, and point to a role for internal timing deficits as a mechanistic explanation for external cues becoming a possible source of self-body input.

Influence of the geometry of an immersed steel workpiece on mass transfer coefficient in a chemical heat treatment fluidised bed

The mass transfer during carburising in a fluidised bed and in a steel workpiece has been studied experimentally in this work. This involved carburising experiment in an electrically heated fluidised bed at 900–970°C with natural gas and air as the atmosphere. A steel workpiece was designed to provide a range of carbon transfer surfaces of different geometries in the fluidised bed, and the carbon transfer coefficient was measured at these surfaces. The carbon transfer coefficient was determined from the carbon distribution within the diffusion layer of the sample. An empirical relationship of the carbon potential as a function of carburising atmosphere, bed temperature and fluidising velocity was determined, based on the understanding of the mass transfer mechanism and analysis of the experimental results.

Internal auditors' assessment of their contribution to financial statement audits : the relation with audit committee and internal audit function characteristics

This paper examines the relation between audit committee characteristics, internal audit function characteristics and internal auditors' assessment of their contribution to financial statement audits. Using survey data from chief internal auditors of 76 Malaysian publicly-listed firms, we provide evidence of a positive relationship between internal auditors' assessment of their contribution to financial statement audits and three audit committee characteristics: the proportion of independent audit committee members, their knowledge and experience of accounting and auditing, and the extent of audit committee review of internal audit programmes, budget and coordination proposals. Further, a positive relationship is found between internal auditors' evaluation of their contribution to the financial statement audit and internal audit function characteristics including size, prior experience of staff in auditing, time availability and the closeness of the function's relationship with the external auditor. The results indicate that more effective audit committees and well-resourced internal audit units tend to be positively associated with the internal auditors' assessment of their contribution to the external audit.

Measurement of the mass transfer coefficient at workpiece surfaces in heat treatment furnaces

The mass (e.g. carbon) transfer coefficient at a workpiece surface is an important kinetic factor to control the heat treatment process of the workpiece and to evaluate heat treatment equipment. The coefficient can be calculated from the carbon concentration at the surface of a sample carburized in a carburizing furnace for a given time. Two common measurement methods which use a thin plate and employ a component as samples respectively are evaluated and compared for sensitivity and uncertainty. The comparison shows that the use of a component produces higher measurement precision and also has the advantage in measuring the carbon transfer coefficients at different treated positions. This method is then extended and discussed methodologically. Also two equations are proposed to calculate the carbon transfer coefficient and its uncertainty, respectively. This method is also applied to measure the carbon transfer coefficient in a fluidized bed heat treatment furnace.

Pd embedded in porous carbon (Pd@CMK-3) as an active catalyst for Suzuki reactions: accelerating mass transfer to enhance the reaction rate

Heterogeneous catalysts are promising candidates for use in organic reactions due to their advantages in separation, recovery, and environment compatibility. In this work, an active porous catalyst denoted as Pd embedded in porous carbon (Pd@CMK-3) has been prepared by a strategy involving immersion, ammoniahydrolysis, and heating procedures. Detailed characterization of the catalyst revealed that Pd(0) and Pd(II) species co-exist and were embedded in the matrix of the porous carbon (CMK-3). The as-prepared catalyst has shown high activity toward Suzuki reactions. Importantly, if the reaction mixture was homogenized by two minutes of ultrasonication rather than magnetic stirring before heating, the resistance to mass transfer in the pore channels was significantly reduced. As a result, the reactions proceeded more rapidly and a four-fold increase in the turnover frequency (TOF) could be obtained. When the ultrasonication was employed throughout the entire reaction process, the conversion could also exceed 90% even without the protection of inert gas, and although the reaction temperature was lowered to 30 °C. This work provides a method for fabricating highly active porous carbon encapsulated Pd catalysts for Suzuki reactions and proves that the problem of mass transfer in porous catalysts can be conveniently resolved by ultrasonication without any chemical modification being necessary.[Figure not available: see fulltext.] © 2014 Tsinghua University Press and Springer-Verlag Berlin Heidelberg.

Marginal and Internal Discrepancies Related to Margin Design of Ceramic Crowns Fabricated by a CAD/CAM System

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

Development of intraxylary phloem and internal cambium in Ipomoea hederifolia (Convolvulaceae)

V. S. PATIL (Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara-390002 India), K. S. RAO (BRD School of Bioscieces, S. P. University, Vallabh Vidyanagar, India), and K. S. RAJPUT (Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara-390002 India). Development of intraxylary phloem and internal cambium in Ipomoea hederifolia (Convolvulaceae). J. Torrey Bot. Soc. 136: 423-432. 2009-In Ipomoea hederifolia L. (Convolvulaceae), internal/intraxylary phloem originated as isolated strands from the procambially derived cells after the formation of protoxylem and protophloem. Bands of internal phloem were apparent in the sixth internode after the development of metacambium. In the relatively thick stems several small arcs/segments of internal cambium ensues from the parenchyma cells between the protoxylem and internal protophloem. Though all the segments were active, some of them (two of them located opposite to each other) were relatively more active. Bidirectional differentiation of these segments gave rise to secondary xylem centrifugally and secondary phloem centripetally, resulting inverted vascular bundles. Rest of the internal cambium segments were unidirectional and formed only secondary phloem centripetally. Like external vascular cambium, the internal cambium was non-storied. Structurally, secondary xylem and phloem was composed of axial and radial system in which rays were mostly uni- to biseriate. Secondary xylem produced by the internal cambium was more or less similar to the xylem formed by the external successive cambia. Secondary phloem produced by the internal cambium was composed of sieve tubes, companion cells, axial and ray parenchyma cells. Simple sieve plates of internal phloem were mostly arranged on transverse end walls in contrast to compound and obliquely placed sieve plates of external phloem formed by the successive cambia.

Reactores aeróbios de lecho fluidizado trifásico con circulación interna: Caracterización hidrodinámica y del soporte

Aerobic internal-loop reactors use active biomass attached in a supporting media (biofilm) with the advantage of retaining a big biomass concentration in a small physical space, removing carbonaceous matter and nitrogen in only one reactor. Liquid circulation occurs due to hydrostatic pressure difference produced by air injection in the riser. In biphasic conditions liquid circulation velocities, gas holdup and oxygen transfer coefficient in four different reactor configurations were studied. For the three-phase conditions, the same parameters in just one of those configurations were evaluated. Also, there were three granular supporting media characterized. On the other hand, the relationship between internal and external tube areas and supporting media concentrations influence the liquid velocity, gas holdup and oxygen mass transfer values and some important supporting media characteristics were observed and compared.

A mass transfer model applied to the supercritical extraction with CO2 of curcumins from turmeric rhizomes (Curcuma longa L)

ABSTRACT: Increasing restrictions on the use of artificial pigments in the food industry, imposed by the international market, have increased the importance of raw materials containing natural pigments. Of those natural substances with potential applications turmeric rhizomes (Curcuma longa L), are one of the most important natural sources of yellow coloring. Three different pigments (curcumin, desmetoxycurcumin, and bis-desmetoxycurcumin) constitute the curcuminoids. These pigments are largely used in the food industry as substitutes for synthetic dyes like tartrazin. Extraction of curcuminoids from tumeric rhizomes with supercritical CO2 can be applied as an alternative method to obtain curcuminoids, as natural pigments are in general unstable, and hence degrade when submitted to extraction with organic solvents at high temperatures. Extraction experiments were carried out in a supercritical extraction pilot plant at pressures between 25 and 30 MPa and a temperature of 318 K. The influence of drying pretreatment on extraction yield was evaluated by analyzing the mass transfer kinetics and the content of curcuminoids in the extracts during the course of extraction. The chemical identification of curcuminoids in both the extract and the residual solid was performed by spectrophotometry. Mass transfer within the solid matrix was described by a linear first-order desorption model, while that in the gas phase was described by a convective mass transfer model. Experimental results showed that the concentration profile for curcuminoids during the supercritical extraction process was higher when the turmeric rhizomes were submitted to a drying pretreatment at 343 K.

Assembly and characterization of supramolecular architectures for biosensor applications

The research has included the efforts in designing, assembling and structurally and functionally characterizing supramolecular biofunctional architectures for optical biosensing applications. In the first part of the study, a class of interfaces based on the biotin-NeutrAvidin binding matrix for the quantitative control of enzyme surface coverage and activity was developed. Genetically modified ß-lactamase was chosen as a model enzyme and attached to five different types of NeutrAvidin-functionalized chip surfaces through a biotinylated spacer. All matrices are suitable for achieving a controlled enzyme surface density. Data obtained by SPR are in excellent agreement with those derived from optical waveguide measurements. Among the various protein-binding strategies investigated in this study, it was found that stiffness and order between alkanethiol-based SAMs and PEGylated surfaces are very important. Matrix D based on a Nb2O5 coating showed a satisfactory regeneration possibility. The surface-immobilized enzymes were found to be stable and sufficiently active enough for a catalytic activity assay. Many factors, such as the steric crowding effect of surface-attached enzymes, the electrostatic interaction between the negatively charged substrate (Nitrocefin) and the polycationic PLL-g-PEG/PEG-Biotin polymer, mass transport effect, and enzyme orientation, are shown to influence the kinetic parameters of catalytic analysis. Furthermore, a home-built Surface Plasmon Resonance Spectrometer of SPR and a commercial miniature Fiber Optic Absorbance Spectrometer (FOAS), served as a combination set-up for affinity and catalytic biosensor, respectively. The parallel measurements offer the opportunity of on-line activity detection of surface attached enzymes. The immobilized enzyme does not have to be in contact with the catalytic biosensor. The SPR chip can easily be cleaned and used for recycling. Additionally, with regard to the application of FOAS, the integrated SPR technique allows for the quantitative control of the surface density of the enzyme, which is highly relevant for the enzymatic activity. Finally, the miniaturized portable FOAS devices can easily be combined as an add-on device with many other in situ interfacial detection techniques, such as optical waveguide lightmode spectroscopy (OWLS), the quartz crystal microbalance (QCM) measurements, or impedance spectroscopy (IS). Surface plasmon field-enhanced fluorescence spectroscopy (SPFS) allows for an absolute determination of intrinsic rate constants describing the true parameters that control interfacial hybridization. Thus it also allows for a study of the difference of the surface coupling influences between OMCVD gold particles and planar metal films presented in the second part. The multilayer growth process was found to proceed similarly to the way it occurs on planar metal substrates. In contrast to planar bulk metal surfaces, metal colloids exhibit a narrow UV-vis absorption band. This absorption band is observed if the incident photon frequency is resonant with the collective oscillation of the conduction electrons and is known as the localized surface plasmon resonance (LSPR). LSPR excitation results in extremely large molar extinction coefficients, which are due to a combination of both absorption and scattering. When considering metal-enhanced fluorescence we expect the absorption to cause quenching and the scattering to cause enhancement. Our further study will focus on the developing of a detection platform with larger gold particles, which will display a dominant scattering component and enhance the fluorescence signal. Furthermore, the results of sequence-specific detection of DNA hybridization based on OMCVD gold particles provide an excellent application potential for this kind of cheap, simple, and mild preparation protocol applied in this gold fabrication method. In the final chapter, SPFS was used for the in-depth characterizations of the conformational changes of commercial carboxymethyl dextran (CMD) substrate induced by pH and ionic strength variations were studied using surface plasmon resonance spectroscopy. The pH response of CMD is due to the changes in the electrostatics of the system between its protonated and deprotonated forms, while the ionic strength response is attributed from the charge screening effect of the cations that shield the charge of the carboxyl groups and prevent an efficient electrostatic repulsion. Additional studies were performed using SPFS with the aim of fluorophore labeling the carboxymethyl groups. CMD matrices showed typical pH and ionic strength responses, such as high pH and low ionic strength swelling. Furthermore, the effects of the surface charge and the crosslink density of the CMD matrix on the extent of stimuli responses were investigated. The swelling/collapse ratio decreased with decreasing surface concentration of the carboxyl groups and increasing crosslink density. The study of the CMD responses to external and internal variables will provide valuable background information for practical applications.

Ocean acidification alleviates low-temperature effects on growth and photosynthesis of the red alga Neosiphonia harveyi (Rhodophyta)

This study aimed to examine interactive effects between ocean acidification and temperature on the photosynthetic and growth performance of Neosiphonia harveyi. N. harveyi was cultivated at 10 and 17.5 °C at present (~380 µatm), expected future (~800 µatm), and high (~1500 µatm) pCO2. Chlorophyll a fluorescence, net photosynthesis, and growth were measured. The state of the carbon-concentrating mechanism (CCM) was examined by pH-drift experiments (with algae cultivated at 10 °C only) using ethoxyzolamide, an inhibitor of external and internal carbonic anhydrases (exCA and intCA, respectively). Furthermore, the inhibitory effect of acetazolamide (an inhibitor of exCA) and Tris (an inhibitor of the acidification of the diffusive boundary layer) on net photosynthesis was measured at both temperatures. Temperature affected photosynthesis (in terms of photosynthetic efficiency, light saturation point, and net photosynthesis) and growth at present pCO2, but these effects decreased with increasing pCO2. The relevance of the CCM decreased at 10 °C. A pCO2 effect on the CCM could only be shown if intCA and exCA were inhibited. The experiments demonstrate for the first time interactions between ocean acidification and temperature on the performance of a non-calcifying macroalga and show that the effects of low temperature on photosynthesis can be alleviated by increasing pCO2. The findings indicate that the carbon acquisition mediated by exCA and acidification of the diffusive boundary layer decrease at low temperatures but are not affected by the cultivation level of pCO2, whereas the activity of intCA is affected by pCO2. Ecologically, the findings suggest that ocean acidification might affect the biogeographical distribution of N. harveyi.