Advances in (poly)phenol research: novel sources, bioavailability, bioaccumulation and bioactivity

In the last decades, increasing scientific evidence has correlated the regular consumption of (poly)phenol-rich foods to a potential reduction of chronic disease incidence and mortality. However, epidemiological evidence on the role of (poly)phenol intake against the risk of some chronic diseases is promising, but not conclusive. In this framework a proper approach to (poly)phenol research is requested, using a step by step strategy. The plant kingdom produces an overwhelming array of structurally diverse secondary metabolites, among which flavonoids and related phenolic and (poly)phenolic compounds constitute one of the most numerous and widely distributed group of natural products. To date, more than 8000 structures have been classified as members of the phytochemical class of (poly)phenol, and among them over 4000 flavonoids have been identified. For this reason, a detailed food (poly)phenolic characterization is essential to identify the compounds that will likely enter the human body upon consumption, to predict the metabolites that will be generated and to unravel the potential effects of phenolic rich food sources on human health. In the first part of this work the attention was focused on the phenolic characterization of fruit and vegetable supplements, considering the increasing attention recently addressed to the so called "nutraceuticals", and on the main coffee industry by-product, namely coffee silverskin. The interest oriented toward (poly)phenols is then extended to their metabolism within the human body, paramount in the framework of their putative health promoting effects. Like all nutrients and non-nutrients, once introduced through the diet, (poly)phenols are subjected to an intense metabolism, able to convert the native compounds into similar conjugated, as well as smaller and deeply modified molecules, which in turn could be further conjugated. Although great strides have been made in the last decades, some steps of the (poly)phenol metabolism remain unclear and are interesting points of research. In the second part of this work the research was focused on a specific bran fraction, namely aleurone, added in feed pellets and in bread to investigate the absorption, metabolism and bioavailability of its phenolic compounds in animal and humans, with a preliminary in vitro step to determine their potential bioaccesibility. This part outlines the best approaches to assess the bioavailability of specific phenolics in several experimental models. The physiological mechanisms explaining the epidemiological and observational data on phenolics and health, are still far from being unraveled or understood in full. Many published results on phenolic actions at cell levels are biased by the fact that aglycones or native compounds have been used, not considering the previously mentioned chemical and biological transformations. In the last part of this thesis work, a new approach in (poly)phenol bioactivity investigation is proposed, consisting of a medium-long term treatment of animals with a (poly)phenol source, in this specific case resveratrol, the detection of its metabolites to determine their possible specific tissue accumulation, and the evaluation of specific parameters and/or mechanism of action at target tissue level. To conclude, this PhD work has contributed to advancing the field, as novel sources of (poly)phenols have been described, the bioavailability of (poly)phenols contained in a novel specific bran fraction used as ingredient has been evaluated in animal and in humans, and, finally, the tissue accumulation of specific (poly)phenol metabolites and the evaluation of specific parameters and/or mechanism of action has been carried out. For these reasons, this PhD work should be considered an example of adequate approach to the investigation of (poly)phenols and of their bioactivity, unavoidable in the process of unequivocally defining their effects on human health.

Production of renewable phenolic resins by thermochemical conversion of biomass: a review

This review covers the production and utilisation of liquids from the thermal processing of biomass and related materials to substitute for synthetic phenol and formaldehyde in phenol formaldehyde resins. These resins are primarily employed in the manufacture of wood panels such as plywood, MDF, particle-board and OSB. The most important thermal conversion methods for this purpose are fast pyrolysis and vacuum pyrolysis, pressure liquefaction and phenolysis. Many feedstocks have been tested for their suitability as sources of phenolics including hard and softwoods, bark and residual lignins. Resins have been prepared utilising either the whole liquid product, or a phenolics enriched fraction obtained after fractional condensation or further processing, such as solvent extraction. None of the phenolics production and fractionation techniques covered in this review are believed to allow substitution of 100% of the phenol content of the resin without impacting its effectiveness compared to commercial formulations based on petroleum derived phenol. This survey shows that considerable progress has been made towards reaching the goal of a price competitive renewable resin, but that further research is required to meet the twin challenges of low renewable resin cost and satisfactory quality requirements. Particular areas of concern are wood panel press times, variability of renewable resin properties, odour, lack of reactive sites compared to phenol and potential for increased emissions of volatile organic compounds.

Pyrolysis and gasification of biomass and acid hydrolysis residues

This research was carried for an EC supported project that aimed to produce ethyl levulinate as a diesel miscible biofuel from biomass by acid hydrolysis. The objective of this research was to explore thermal conversion technologies to recover further diesel miscible biofuels and/or other valuable products from the remaining solid acid hydrolysis residues (AHR). AHR consists of mainly lignin and humins and contains up to 80% of the original energy in the biomass. Fast pyrolysis and pyrolytic gasification of this low volatile content AHR was unsuccessful. However, successful air gasification of AHR gave a low heating value gas for use in engines for power or heat with the aim of producing all the utility requirements in any commercial implementation of the ethyl levulinate production process. In addition, successful fast pyrolysis of the original biomass gave organic liquid yields of up to 63.9 wt.% (dry feed basis) comparable to results achieved using a standard hardwood. The fast pyrolysis liquid can be used as a fuel or upgraded to biofuels. A novel molybdenum carbide catalyst was tested in fast pyrolysis to explore the potential for upgrading. Although there was no deoxygenation, some bio-oil properties were improved including viscosity, pH and homogeneity through decreasing sugars and increasing furanics and phenolics. AHR gasification was explored in a batch gasifier with a comparison with the original biomass. Refractory and low volatile content AHR gave relatively low gas yields (74.21 wt.%), low tar yields (5.27 wt.%) and high solid yields (20.52 wt.%). Air gasification gave gas heating values of around 5MJ/NM3, which is a typical value, but limitations of the equipment available restricted the extent of process and product analysis. In order to improve robustness of AHR powder for screw feeding into gasifiers, a new densification technique was developed based on mixing powder with bio-oil and curing the mixture at 150°C to polymerise the bio-oil.

In situ catalytic upgrading of bio-oil using supported molybdenum carbide

The aim of this work is to improve some of the less desirable properties of bio-oil via the catalytic fast pyrolysis of sugarcane bagasse using a novel supported molybdenum carbide (20 wt.% MoC/AlO ) catalyst. Proximate and elemental analysis of the bagasse were carried out to determine the moisture, ash, carbon, hydrogen, nitrogen and oxygen content. The ground pellets were classified in sieves to a size range of 0.25-1 mm and were pyrolysed in a 300 g h fluidised bed reactor at 500 C. MoC/AlO replaced the sand in the fluidised bed reactor in different proportions (0 wt.%, 12 wt.%, 25 wt.% and 50 wt.%) to investigate the effect of this catalyst on the pyrolysis products. Bio-oil yield results showed that ground sugarcane bagasse pellets gave high organic yields in the bio-oil of 60.5 wt.% on dry feed with a total liquid yield of 73.1 wt.% on dry feed without catalyst. Increasing the catalyst proportions in the fluidised bed reduced bio-oil yields, significantly reduced sugars (as a-levoglucosan) concentration and increased furanics and phenolics concentration in the bio-oil. It was observed that the higher the concentration of the 20 wt.% MoC/AlO catalyst in the fluidised bed the lower the viscosity of the bio-oil. © 2013 Elsevier B.V. All rights reserved.

The ethnopharmacology of plants used as both food and medicine in a Totonac community of Mexico

Most ethnopharmacological studies overlook food plants, yet many edible plants, also have medicinal value. I documented plants that are used as both food and medicine by the Totonac of Zapotitlan de Mendez, Mexico and recorded the presence of selected secondary compounds, and physical characteristics in these plants. Photoactivity, antimicrobial, and antifungal assays also were performed. The presence of these properties were compared among food/medicine plants, food, medicinal, and randomly selected plants. I predicted that a higher percentage of medicinal plants would contain the secondary compounds, physical characteristics, and bioactivity compared to the other groups. Phenolics and cyanogenic glycosides in the medicinal group were significantly greater than in the food/medicine group. The food plants did not differ greatly from the medicinal plants. This research indicates that including food plants in ethnomedical studies could provide a more complete knowledge of peoples therapeutic resources and practices. ^

Ocean acidification and the loss of phenolic substances in marine plants

Rising atmospheric CO2 often triggers the production of plant phenolics, including many that serve as herbivore deterrents, digestion reducers, antimicrobials, or ultraviolet sunscreens. Such responses are predicted by popular models of plant defense, especially resource availability models which link carbon availability to phenolic biosynthesis. CO2 availability is also increasing in the oceans, where anthropogenic emissions cause ocean acidification, decreasing seawater pH and shifting the carbonate system towards further CO2 enrichment. Such conditions tend to increase seagrass productivity but may also increase rates of grazing on these marine plants. Here we show that high CO2 / low pH conditions of OA decrease, rather than increase, concentrations of phenolic protective substances in seagrasses and eurysaline marine plants. We observed a loss of simple and polymeric phenolics in the seagrass Cymodocea nodosa near a volcanic CO2 vent on the Island of Vulcano, Italy, where pH values decreased from 8.1 to 7.3 and pCO2 concentrations increased ten-fold. We observed similar responses in two estuarine species, Ruppia maritima and Potamogeton perfoliatus, in in situ Free-Ocean-Carbon-Enrichment experiments conducted in tributaries of the Chesapeake Bay, USA. These responses are strikingly different than those exhibited by terrestrial plants. The loss of phenolic substances may explain the higher-than-usual rates of grazing observed near undersea CO2 vents and suggests that ocean acidification may alter coastal carbon fluxes by affecting rates of decomposition, grazing, and disease. Our observations temper recent predictions that seagrasses would necessarily be "winners" in a high CO2 world.

Seawater carbonate chemistry and Celleporella hyalina biological processes during experiments, 2011

Increased anthropogenic CO2 emissions in the last two centuries have lead to rising sea surface temperature and falling ocean pH, and it is predicted that current global trends will worsen over the next few decades. There is limited understanding of how genetic variation among individuals will influence the responses of populations and species to these changes. A microcosm system was set up to study the effects of predicted temperature and CO2 levels on the bryozoan Celleporella hyalina. In this marine species, colonies grow by the addition of male, female and feeding modular individuals (zooids) and can be physically subdivided to produce a clone of genetically identical colonies. We studied colony growth rate (the addition of zooids), reproductive investment (the ratio of sexual to feeding zooids) and sex ratio (male to female zooids) in four genetically distinct clonal lines. There was a significant effect of clone on growth rate, reproductive investment and sex ratio, with clones showing contrasting responses to the various temperature and pH combinations. Overall, decreasing pH and increasing temperature caused reduction of growth, and eventual cessation of growth was often observed at the highest temperature, especially during the latter half of the 15-day trials. Reproductive investment increased with increasing temperature and decreasing pH, varying more widely with temperature at the lowest pH. The increased production of males, a general stress response of the bryozoan, was seen upon exposure to reduced pH, but was not expressed at the highest temperature tested, presumably due to the frequent cessation of growth. Further to the significant effect of pH on the measured whole-colony parameters, observation by scanning electron microscopy revealed surface pitting of the calcified exoskeleton in colonies that were exposed to increased acidity. Studying ecologically relevant processes of growth and reproduction, we demonstrate the existence of relevant levels of variation among genetic individuals which may enable future adaptation via non-mutational natural selection to falling pH and rising temperature.

Ferulates and lignin structural composition in cork

The structure of lignin and suberin, and ferulic acid (FA) content in cork from Quercus suber L. were studied. Extractive-free cork (Cork), suberin, desuberized cork (Cork(sap)), and milled-cork lignins (MCL) from Cork and Cork(sap) were isolated. Suberin composition was determined by GC-MS/FID, whereas the polymers structure in Cork, Corksap, and MCL was studied by Py-TMAH and 2D-HSQC-NMR. Suberin contained 94.4% of aliphatics and 3.2% of phenolics, with 90% of omega-hydroxyacids and alpha,omega-diacids. FA represented 2.7% of the suberin monomers, overwhelmingly esterified to the cork matrix. Py-TMAH revealed significant FA amounts in all samples, with about 3% and 6% in cork and cork lignins, respectively. Py-TMAH and 2D-HSQC-NMR demonstrated that cork lignin is a G-lignin (>96% G units), with a structure dominated by beta-O-4' alkyl-aryl ether linkages (80% and 77% of all linkages in MCL and MCLsap, respectively), followed by phenylcoumarans (18% and 20% in MCL and MCLsap, respectively), and smaller amounts of resinols (ca. 2%) and dibenzodioxocins (1%). HSQC also revealed that cork lignin is heavily acylated (ca. 50%) exclusively at the side-chain gamma-position. Ferulates possibly have an important function in the chemical assembly of cork cell walls with a cross-linking role between suberin, lignin and carbohydrates.

Características fitoquímicas e propriedades antioxidantes do grão de café verde

Entre os produtos comercializados a nível mundial, o café é um dos mais valiosos, sendo apenas superado pelo petróleo em termos de movimentações financeiras. Assim, torna-se indiscutível a importância do café na política e na economia de muitos países, pois o seu cultivo, processamento, comercialização, transporte e mercado criam milhões de empregos por todo o mundo. A composição química do café constitui um parâmetro fundamental na distinção das diferentes variedades deste produto, recorrendo-se frequentemente à análise de compostos como a cafeína, o ácido hidroxicinâmico e o ácido clorogénico, entre outros. O principal objetivo deste trabalho focou-se no estudo do perfil dos compostos bioativos presentes em grãos de café verde provenientes de Cabo Verde e das inerentes propriedades antioxidantes. Mais concretamente, procedeu-se à quantificação dos compostos bioativos com maior importância no café, como é o caso dos fenólicos e flavonoides totais, das antocianinas e dos carotenoides. A quantidade de compostos fenólicos totais encontrados na amostra dos grãos de café verde foi 4,855 mg/g. Quanto à quantidade de flavonoides totais, foi 41,2 mg/g e de antocianinas 0,465 mg/g. Relativamente aos carotenoides estudados, a clorofila a apresenta-se em quantidade igual a 1,5×10 mg/g, a clorofila b, 1,6×10 mg/g, o licopeno, 6×10 mg/g e o β-caroteno aparenta ser inexistente nesta amostra. A quantificação destes compostos bioativos comprovou a sua presença na amostra de café verde e, consequentemente, evidenciou os potenciais benefícios que este produto traz para a saúde, sendo a capacidade antioxidante o mais prevalente.

Seleção de jabuticabeiras juvenis considerando o vigor, o potencial antioxidante e a tolerância a geadas

The jabuticaba tree has great potential for commercial exploitation. However, its is very little used. This fact shows to be necessary to do studies that allow understand their growth behavior during the year and, if it is tolerant to frost. So that it can establish management strategies for cultivation in orchard. Other point, it is the fact that the long juvenile period of jabuticaba tree limits its use. However, many species have compound leaves that characterize them as functional compounds, what to posible its commercialization. If the leaf jabuticaba tree also present such nutraceutical compounds, this it may become an alternative source of income until the plant to start its yield. The objectives of this study were to analyze the growth behavior, the occurrence of flowering and fruit set, and the frost tolerance of jabuticaba tree genotypes present in the collection of Native Fruit from UTFPR – Câmpus Dois Vizinhos. Associated growth analysis was made evaluation of genetic divergence among these genotypes, checking the adaptive behavior in orchard condition through adaptability and stability analysis based on growth measures to stem and shoots; estimating the repeatability coefficient of stem length of characters and primary shoots, and determine the minimum number of evaluations able to provide certain levels of prediction of the actual value of these individuals. Also determined the genetic divergence among genotypes as the leaves of antioxidant activity by DPPH and ABTS methods, as well as the determination of total phenolics. The genotypes studied were put in orchard in 2009. The growth response in the three cycles was variable between months and genotypes, what it can be difficult the practices in the orchard if it do not use clones. Genotypes 'Silvestre' and 'Açú' showed greater width and leaf area compared with other genotypes, but such behavior is not favored for increased stem growth and primary shoots. Foliar increments in most genotypes occurred in the fall for leaf width, spring for length and leaf area, despite the winter also arise with genotypes, it showed superiority to width and leaf area. Most jabuticabas trees were juvenile stage with only four starting at its transition between the vegetative and reproductive phase. Tolerance to frost was observed in 26 families jabuticabeira of the 29 present in the collection. The diversity among the genotypes was to change with the time, already in each cycle, there was the formation of different groups by the methods used. The methods tested for adaptability and stability of the jabuticaba tree growth behavior did not show the same pattern in the results. The number of measurements needed to predict the actual value of genotypes based on variables evaluated was approximately one to the stem length and four for the shoots based on the method of main components of covariance with 90% probability. he antioxidant activity of the extracts of leaves of jabuticaba tree genotypes were demonstrated high when compared to other species by methods DPPH and ABTS, as well as the amount of phenolic compounds. Genotype 'Silvestre' and 'IAPAR' showed the highest antioxidant activity in the leaves. However, the genetic divergence among genotypes jabuticaba tree from collection of Native Fruit trees at UTFPR - Câmpus Dois Vizinhos for antioxidant activity leaves showed that they have great homogeneity among them and the low divergence. However, it is recommended as possible hybridization the use as parents, José 4, IAPAR 4 and Fernando Xavier genotypes.

Functional studies of lignin biosynthesis genes and putative flowering gene in Miscanthus x giganteus and studies on indolyl glucosinolate biosynthesis and translocation in Brassica oleracea

The first topic area of this thesis involved studies on the accumulation and translocation of glucosinolates (GSs), bioactive secondary plant compounds, in broccoli plants. Changes in GS accumulation and gene expression levels in response to exogeneous methyl jasmonate (MeJA) treatment were analyzed in different tissue types at different developmental stages of broccoli. Greater accumulation of GSs with MeJA treatment was observed in apical leaves of broccoli seedlings and florets of plants at harvest maturity. Increases in indolyl GS in apical leaves of seedlings and florets were coupled with the up-regulation of indolyl GS biosynthesis genes. The accumulation of indolyl GSs appears to be modulated by MeJA treatment in an organ-specific manner for optimal distribution of defense substances in the plant. Metabolic profiling of hydrophilic metabolites using GC-MS demonstrated increased accumulation of various phenolics, ascorbates and amino acids in broccoli tissues after MeJA treatment. Distinct changes in carbohydrate levels observed between different tissues (vegetative leaves and floret tissues) of broccoli plants after treatment suggest that carbon metabolism is differentially modulated by MeJA treatment in different tissue types depending on sink-source relationships. Reduced levels of hexose sugars and tricarboxylic acid intermediates after MeJA treatment may reflect the increased requirement for carbon and energy needed to drive secondary product biosynthesis to accumulate metabolites for defense against insects and other herbivores. Substantial increases of indolyl and aromatic GSs after exogenous treatment with MeJA in stem and petioles of seedlings and the existence of intact indolyl-GS forms in phloem exudates suggest enhanced de novo synthesis in combination with active transport. Indoly GSs share structural similarities with the auxin, IAA, and may interact with components of the auxin transport system for intra- and extra-cellular transport or translocation. Application of the auxin efflux inhibitor, 1-naphthylphthalamic acid (NPA) reduced MeJA-mediated accumulation of indolyl GSs in broccoli florets and seedling tissues. NPA did not inhibit expression of indolyl GS biosynthesis genes shown to be upregulated by MeJA treatment or the accumulation of tryptophan, the amino acid precursor of indolyl GSs. Exogenous application of benzyl GS to Arabidopsis roots induced ectopic expression of the PIN1 protein associated with the auxin transport system similar to treatment with NPA, again suggesting GS interaction with the auxin efflux carrier system. The inhibitory effect of NPA on MeJA-mediated accumulation of GS may be due to competitive binding of NPA to auxin efflux carrier components and that GS transport is mediated by the auxin transport system. The inhibitory effect of NPA on indolyl and aromatic GS accumulation and the bioactivity of exogenous treatment of these GS compounds in PIN1 localization, Arabidopsis root growth, and gravitrophic response suggest that indolyl and aromatic GSs may be antagonistic to IAA transport and biosynthesis. Indolyl and aromatic GSs can also be potentially converted into IAA by hydrolysis. This intrinsic feature of GSs may be the part of a sophisticated regulatory process where the metabolic pathways in the plant shift from active growth to a reversible defense posture in response to biotic or abiotic stress. It seems likely that indolyl and aromatic GSs are important compounds that provide connections between jasmonate and auxin signaling. Further studies are required to reveal the regulatory mechanism for crosstalk between the two hormones. The third part of this research was to investigate effect of selenium fertilization and MeJA treatment on accumulation of GSs in broccoli florets. Increasing dietary intake of the element selenium (Se) has been shown to reduce the risk of cancer. Simultaneous enhancement of both Se and GS concentrations in broccoli floret tissue were conducted through the combined treatment of MeJA with Se fertilization. A low level of Se fertilization (concentration) with MeJA treatment displayed no significant changes in total aliphatic GS concentrations with 90% and 50% increases in indolyl and total GSs concentrations, respectively. This result suggests that Se- and GS-enriched broccoli with improved health-promoting properties can be generated by this combined treatment. The second topic of this thesis was conducted to provide basic information required to improve biomass quality and productivity and develop tools for gene transformation in Miscanthus x giganteus. The perennial rhizomatous grass, Miscanthus x giganteus is an ideal biomass crop due to its rapid vegetative growth and high biomass yield potential. As a naturally occurring sterile hybrid, M. x giganteus must be propagated vegetatively by mechanicalling divided rhizomes or from micropropagated plantlets. The effect of callus type, age and culture methods on regeneration competence was studied to improve regeneration efficiency and shorten the period of tissue culture in M. x giganteus propagation. Seven lignin biosynthesis genes and one putative flowering gene were isolated from M. x giganteus by PCR reactions using maize othologous sequences. Southern hybridization and nuclear DNA content analysis indicated that the genes isolated from M. x giganteus exist in the genome of other Miscanthus species as multiple copies. Analysis of lignin content and histological staining of lignin deposition indicated that higher lignin content is found in mature stem node tissues compared to young leaves and apical stem nodal tissues. Cell wall lignification is associated with increasing tissue maturity in Miscanthus species. RNAi and antisense constructs harboring sequences of these genes were developed to generate Miscanthus transgenic plants with suppressed of lignin biosynthesis and delayed flowering.

Potencial antifúngico, antioxidante e inibidor da produção de aflatoxina por extratos fenólicos de Chlorella sp. e Spirulina platensis

A contaminação fúngica acarreta alterações na qualidade nutricional e no valor econômico de produtos alimentícios podendo causar danos patológicos em plantas, animais e humanos. A identificação da atividade antioxidante, antifúngica e antimicotoxinas, em extratos de microalgas com propriedade de inibir a multiplicação de fungos e subseqüente produção de micotoxinas abre a perspectiva de empregar substâncias mais eficientes e com maior ação específica contra estes microorganismos. Entre os compostos com propriedades inibidoras de radicais livres, de crescimento fúngico e produção de micotoxinas, destacam-se os compostos fenólicos, que podem inibir a atividade metabólica microbiana, dificultando a atividade de enzimas. Neste estudo foram avaliados o poder de inibição de multiplicação fúngica de Rhizopus oryzae e Aspergillus flavus pelos extratos fenólicos de Chlorella sp. e Spirulina platensis, bem como sua atividade antioxidante, e a atividade antimicotoxinas da última microalga contra Aspergillus flavus. O conteúdo de fenóis totais foi em média 1000 µgfenóis/g Spirulina platensis e 600 µgfenóis/g Chlorella sp., sendo que o acido gálico e o cafeíco foram identificados como compostos majoritários na Spirulina platensis. As determinações de glicosamina (parede celular) e ergosterol (membrana celular) mostraram-se bons indicativos do desenvolvimento microbiano permitindo uma boa estimativa da inibição dele. O extrato fenólico de Spirulina platensis apresentou capacidade de inibir cerca de 50% a formação da parede e da membrana celular para ambos os fungos estudados e de 100% a produção de aflatoxina B1 até o 10º dia de cultivo do Aspergillus flavus. Além disso, o extrato metanólico de Spirulina platensis inativou 53,5% o DPPH reativo, limitou o escurecimento enzimático ocasionado pela peroxidase em 55% e inibiu a peroxidação lipídica em 46% após 14 dias de armazenamento sob luz. Estes resultados mostram que a ação antifúngica, antimicotoxinas e antioxidante está naturalmente presente em alguns tecidos microbianos e que encontrar a forma de extraí-los e aplicá-los como conservantes alimentícios é muito promissor para substituição aos antifúngicos e outros conservantes químicos.

Validação de metodologia analítica por CLAE-DAD para determinação de compostos fenólicos e atividade biológica em coprodutos agroindustriais

The southern region of Brazil, especially the states of Parana and Santa Catarina stand out for growing grapes and apples for fresh consumption and in order to add value to these products, process the material for the production of wine, juices and jellies . As a result large quantities of by-products, such as peels, seeds and pulp are produced becoming environmental problems. Studies reuse of these by-products have attracted interest because they have shown a high biological potential, due to the presence of high levels of phenolic compounds, which are associated with a lower incidence of disease caused by oxidative stress, due to its antioxidant, antiinflammatory and antibacterial properties. Currently, few studies are presented on the phenolic composition and biological potential of waste grape variety Bordô (Vitis labrusca) and apple (Malus domestica) Gala variety, cultivated in southern Brazil. Within this context, the objectives of this study were: compare the efficiency of solidliquid and liquid-liquid extraction, perform the optimization and validation of analytical methodology by HPLC-DAD for the separation, identification and quantification of multiclass phenolic compounds, evaluate the activity antioxidant by sequestering methods of free radical 2,2-diphenyl-1 picrilhidrazina (DPPH) and 2,2-azino-bis (3- ethyl-benzthiazoline-6-sulphonic acid) (ABTS) solution, reduction of Fe3+ in Fe2+ method (FRAP), ORAC, RP-HPLC-ABTS online, Rancimat and determination of total phenolics three agro-industrial byproducts, pomace and stems grape Bordô produced in Paraná Southwest region and Gala apple pomace coming from the Santa Catarina West. Optimization and validation of chromatographic method showed satisfactory quality parameters for the compounds of interest and the solidliquid extraction was more efficient in extracting phenolic evaluated. The three byproducts evaluated showed significant levels of phenolic compounds when analyzed by HPLC, especially flavonoids, catechin and epicatechin besides that showed significant antioxidant capacity. The grape stems extract had the highest sequestration capacity of DPPH and ABTS radical and reduced iron, and high content of phenolic compounds. The apple pomace extract showed the best response to the Rancimat method, which indicates a high potential to protect the oil from lipid oxidation, was no significant difference when compared to synthetic antioxidant TBHQ. The results of this study showed that the agro-industrial coproducts analyzed are rich in phenolic compounds of high antioxidant capacity and therefore must be better explored by the food and pharmaceutical industries.

Estudo das propriedades físico-químicas e avaliação de compostos bioativos em camarinhas ('Corema album' L.D. Don)

As camarinhas ('Corema album' L. D. Don) são pequenos frutos selvagens que se desenvolvem em arbustos dunares ao longo das costas da Península Ibérica. Apesar de serem ainda pouco conhecidas, as camarinhas são pequenos frutos que podem pertencer ao vasto grupo dos frutos vermelhos, vulgarmente conhecidos pelos seus efeitos benéficos na saúde. Desta forma, neste estudo foram realizadas várias análises às camarinhas no que respeita a propriedades de natureza física e química e em particular a alguns compostos com efeitos bioativos. Com a realização deste trabalho pretendeu-se avaliar as propriedades físico-químicas das camarinhas, bem como dos compostos bioativos com potenciais benefícios para a saúde. Nesse sentido, as bagas de camarinha foram avaliadas quanto às suas propriedades físicas (dimensões, peso, cor e textura), propriedades químicas (humidade, acidez, ºBrix, fibra, açúcares totais, açúcares redutores e vitamina C) e propriedades fenólicas (compostos fenólicos totais, orto-difenóis, flavonóides, taninos e atividade antioxidante, por DPPH e ABTS), em diferentes extratos de amostras de polpas e de grainhas liofilizadas. Os primeiros extratos ainda foram submetidos a uma simulações das condições do trato digestivo, para avaliar a bioacessibilidade dos compostos fenólicos e da atividade antioxidante. Este trabalho teve ainda como objetivo conhecer a bioacessibilidade dos compostos fenólicos totais e da sua atividade antioxidante, através do método da simulação “in vitro” das diferentes etapas do trato gastrointestinal. No que diz respeito às propriedades físicas analisadas, as camarinhas frescas demonstraram uma altura média de 8,6 mm, um diâmetro médio de 9,4 mm e uma massa média de 0,7 g. Relativamente à caraterização da cor, estas apresentavam uma cor clara (L*=79,8). Quanto à textura apresentaram alguma elasticidade média (2,9 mm) e uma baixa dureza (1,9 N). Nas análises químicas as camarinhas revelaram ser compostas, maioritariamente, por água (87,9%), por açúcares totais e fibras. Para além disso, apresentavam um teor em sólidos solúveis totais de 6,3 ºBrix, uma acidez de 1,4 g ácido tartárico/100 g e um teor de vitamina C de 2,8 mg de ácido ascórbico/100 g.Na quantificação dos compostos fenólicos totais e flavonóides, os extratos de acetona:água das amostras de polpas de camarinha branca apresentaram os valores mais elevados, 1614,1 mg EAG/100 g e 143,7 mg EQ/100 g, respetivamente. Relativamente à quantificação dos orto-difenóis e dos taninos os extratos de metanol e de acetona:água das amostras de grainhas de camarinha branca registaram os valores de 23,4 mg EAG/100 g e 915,7 mg/100 g, respetivamente. Na atividade antioxidante por DPPH e ABTS os extratos de acetona:água das amostras de polpas de camarinha branca apresentaram, respetivamente, os valores de 40,1 µmol ET/g e de 79,6 µmol ET/g. Na avaliação da bioacessibilidade verificou-se que ocorreu uma maior percentagem de compostos fenólicos disponíveis para absorção e uma maior preservação da sua atividade antioxidante nos extratos das grainhas, comparativamente aos extratos das polpas. Com este estudo concluiu-se que as camarinhas são pequenos frutos portadores de um grande potencial em diversos compostos bioativos benéficos para a saúde dos consumidores.

Estudo das propriedades físico-químicas e avaliação de compostos bioativos em 'Physalis peruviana L.'

O rápido crescimento da procura e na produção de Physalis está associado às suas características nutracêuticas e medicinais, por estarem associadas ao bemestar e à saúde. O reconhecimento e o conceito da qualidade dos frutos é cada vezmais importante, abrangendo a aparência do produto, o aroma, a textura e o valor nutricional. Com a realização deste trabalho pretendeu-se avaliar as propriedades físicoquímicas das bagas de physalis, bem como os conteúdos em compostos bioativos com potenciais benefícios para a saúde humana. Neste trabalho foi avaliada uma variedade de physalis (Physalis peruviana L.), sendo os frutos provenientes de uma exploração localizada na região Norte-Centro de Portugal. Foram realizadas análises físicas às 'physalis' frescas, tendo sido determinados o calibre, a cor e a textura. Na análise química, realizaram-se as determinações, da humidade, da fibra bruta, dos açúcares totais e redutores, da acidez total titulável, do teor de sólidos solúveis totais, do ácido ascórbico, dos carotenóides, dos compostos fenólicos totais, dos orto-difenóis e dos flavonóides. Foi ainda determinada a atividade antioxidante pelos métodos ABTS e DPPH e os extratos submetidos a condições simulantes do trato digestivo. As physalis em estudo apresentaram, em média um diâmetro de 1,69 cm e uma massa de 2,77 g. Relativamente à cor, as bagas apresentaram-se claras, (L*=65,72), e com uma tendência para a cor vermelha (a*=16,69), e uma forte intensidade amarela (b*=58,11). No que diz respeito à textura, a firmeza foi de 2,40 N e a elasticidade foi de 2,94 mm. Quanto à caracterização química foram encontrados os seguintes resultados: 83,02% de água, 4,61% de fibra, 8,79% de açúcares totais, 1,25% de acidez total, expressa em ácido cítrico. A amostra continha ainda 5,95 μg/g de carotenoides 26,7 mg de ácido ascórbico por 100 g. Os valores de fenóis totais (42,74 e 59,95 mg EAG/100 g) e de atividade antioxidante determinada pelo método DPPH (7,73 e 9,61 μmol TE/g), e pelo método ABTS (12,28 e 13,71 μmol TE/g) variam de acordo com as condições de extração. Verificou-se ainda uma correlação forte entre os dois métodos. No que diz respeito às condições in vitro de simulação das condições do trato digestivo, verificou-se um decréscimo ao longo do trato digestivo tanto no teor em compostos fenólicos bem como na atividade antioxidante. Em termos globais houve uma retenção de 43% dos compostos fenólicos totais e 26% da atividade antioxidante.