Determination of bioactive and potentially toxic compounds in food: raw material analysis and shelf life evaluation

"Bioactive compounds" are extranutritional constituents that typically occur in small quantities in food. They are being intensively studied to evaluate their effects on health. Bioactive compounds include both water soluble compounds, such as phenolics, and lipidic substances such as n-3 fatty acids, tocopherols and sterols. Phenolic compounds, tocopherols and sterols are present in all plants and have been studied extensively in cereals, nuts and oil. n-3 fatty acids are present in fish and all around the vegetable kingdom. The aim of the present work was the determination of bioactive and potentially toxic compounds in cereal based foods and nuts. The first section of this study was focused on the determination of bioactive compounds in cereals. Because of that the different forms of phytosterols were investigated in hexaploid and tetraploid wheats. Hexaploid cultivars were the best source of esterified sterols (40.7% and 37.3% of total sterols for Triticum aestivum and Triticum spelta, respectively). Significant amounts of free sterols (65.5% and 60.7% of total sterols for Triticum durum and Triticum dicoccon, respectively) were found in the tetraploid cultivars. Then, free and bound phenolic compounds were identified in barley flours. HPLCESI/ MSD analysis in negative and positive ion mode established that barley free flavan-3- ols and proanthocyanidins were four dimers and four trimers having (epi)catechin and/or (epi)gallocatechin (C and/or GC) subunits. Hydroxycinnamic acids and their derivatives were the main bound phenols in barley flours. The results obtained demonstrated that barley flours were rich in phenolic compounds that showed high antioxidant activity. The study also examined the relationships between phenolic compounds and lipid oxidation of bakery. To this purpose, the investigated barley flours were used in the bakery production. The formulated oven products presented an interesting content of phenolic compounds, but they were not able to contain the lipid oxidation. Furthermore, the influence of conventional packaging on lipid oxidation of pasta was evaluated in n-3 enriched spaghetti and egg spaghetti. The results proved that conventional packaging was not appropriated to preserve pasta from lipid oxidation; in fact, pasta that was exposed to light showed a high content of potentially toxic compounds derived from lipid oxidation (such as peroxide, oxidized fatty acids and COPs). In the second section, the content of sterols, phenolic compounds, n-3 fatty acids and tocopherols in walnuts were reported. Rapid analytical techniques were used to analyze the lipid fraction and to characterize phenolic compounds in walnuts. Total lipid chromatogram was used for the simultaneous determination of the profile of sterols and tocopherols. Linoleic and linolenic acids were the most representative n-6 and n-3 essential dietary fatty acids present in these nuts. Walnuts contained substantial amounts of γ- and δ-tocopherol, which explained their antioxidant properties. Sitosterol, Δ5-avenasterol and campesterol were the major free sterols found. Capillary electrophoresis coupled to DAD and microTOF was utilized to determine phenolic content of walnut. A new compound in walnut ((2E,4E)- 8-hydroxy-2,7-dimethyl-2,4-decadiene-1,10-dioic acid 6-O-β-D-glucopiranosyl ester, [M−H]− 403.161m/z) with a structure similar to glansreginins was also identified. Phenolic compounds corresponded to 14–28% of total polar compounds quantified. Aglycone and glycosylated ellagic acid represented the principal components and account for 64–75% of total phenols in walnuts. However, the sum of glansreginins A, B and ((2E,4E)-8-hydroxy- 2,7-dimethyl-2,4-decadiene-1,10-dioic acid 6-O-β-D-glucopiranosyl ester was in the range of 72–86% of total quantified compounds.

Sistema per l'estrazione e purificazione di sostanze antiossidanti naturali nelle acque di vegetazione delle olive

The olive oil extraction industry is responsible for the production of high quantities of vegetation waters, represented by the constitutive water of the olive fruit and by the water used during the process. This by-product represent an environmental problem in the olive’s cultivation areas because of its high content of organic matter, with high value of BOD5 and COD. For that reason the disposal of the vegetation water is very difficult and needs a previous depollution. The organic matter of vegetation water mainly consists of polysaccharides, sugars, proteins, organic acids, oil and polyphenols. This last compounds are the principal responsible for the pollution problems, due to their antimicrobial activity, but, at the same time they are well known for their antioxidant properties. The most concentrate phenolic compounds in waters and also in virgin olive oils are secoiridoids like oleuropein, demethyloleuropein and ligstroside derivatives (the dialdehydic form of elenolic acid linked to 3,4-DHPEA, or p-HPEA (3,4-DHPEA-EDA or p-HPEA-EDA) and an isomer of the oleuropein aglycon (3,4-DHPEA-EA). The management of the olive oil vegetation water has been extensively investigated and several different valorisation methods have been proposed, such as the direct use as fertilizer or the transformation by physico-chemical or biological treatments. During the last years researchers focused their interest on the recovery of the phenolic fraction from this waste looking for its exploitation as a natural antioxidant source. At the present only few contributes have been aimed to the utilization for a large scale phenols recovery and further investigations are required for the evaluation of feasibility and costs of the proposed processes. The present PhD thesis reports a preliminary description of a new industrial scale process for the recovery of the phenolic fraction from olive oil vegetation water treated with enzymes, by direct membrane filtration (microfiltration/ultrafiltration with a cut-off of 250 KDa, ultrafiltration with a cut-off of 7 KDa/10 KDa and nanofiltration/reverse osmosis), partial purification by the use of a purification system based on SPE analysis and by a liquid-liquid extraction system (LLE) with contemporary reduction of the pollution related problems. The phenolic fractions of all the samples obtained were qualitatively and quantitatively by HPLC analysis. The work efficiency in terms of flows and in terms of phenolic recovery gave good results. The final phenolic recovery is about 60% respect the initial content in the vegetation waters. The final concentrate has shown a high content of phenols that allow to hypothesize a possible use as zootechnic nutritional supplements. The purification of the final concentrate have garanteed an high purity level of the phenolic extract especially in SPE analysis by the use of XAD-16 (73% of the total phenolic content of the concentrate). This purity level could permit a future food industry employment such as food additive, or, thanks to the strong antioxidant activity, it would be also use in pharmaceutical or cosmetic industry. The vegetation water depollutant activity has brought good results, as a matter of fact the final reverse osmosis permeate has a low pollutant rate in terms of COD and BOD5 values (2% of the initial vegetation water), that could determinate a recycling use in the virgin olive oil mechanical extraction system producing a water saving and reducing thus the oil industry disposal costs .

From Folke Medicine to Medicine Chemistry: Study, Using in Vitro and Cellular Assays, of Receptors Mechanism Involved in the Activities of Natural Compounds

My Doctorate Research has been focused on the evaluation of the pharmacological activity of a natural extract of chestnut wood (ENC) towards the cardiovascular and gastrointestinal system and on the identification of the active compounds. The ENC has been shown to contain more than 10% (w/w) of phenolic compounds, of which tannins as Vescalgin and Castalgin are the more representative. ENC cardiovascular effects have been investigated in guinea pig cardiac preparations; furthermore its activity has been evalueted in guinea pig aorta strips. ENC induced transient negative chronotropic effect in isolated spontaneously beating right atria and simultaneously positive inotropic effect in left atria driven at 1 Hz. Cardiac cholinergic receptors are not involved in the negative chronotropic effect and positive inotropic effects are not related to adrenergic receptors. In vascular smooth muscle, natural extract of chestnut did not significantly change the contraction induced by potassium (80 mM) or that induced by noradrenaline (1μM). In guinea pig ileum, ENC reduced the maximum response to carbachol in a concentrationdependent manner and behaved as a reversible non competitive antagonist. In guinea pig ileum, the antispasmodic activity of ENC showed a significant antispasmodic activity against a variety of different spasmogenic agents including histamine, KCl, BaCl2. In guinea pig proximal colon, stomach and jejunum, ENC reduced the maximum response to carbachol in a concentrationdependent manner and behaved as a reversible non competitive antagonist. ENC contracted gallbladder guinea pig in a reversible and concentration-dependent manner. This effect does not involve cholinergic and cholecystokinin receptors and it is reduced by nifedipine. ENC relaxed Oddi sphincter smooth muscle. The cholecystokinetic and Oddi sphincter relaxing activities occurred also in guinea pigs fed a lithogenic diet. The cholecystokinetic occurred also in human gallbladder. The Fractionation of the extract led to the identification of the active fraction.

Phenotype and genotype characterization of Monilinia spp. isolates and preformed antifungal compounds in peach peel fruit at different developmental stages

The brown rot fungi belong to a group of fungal pathogens that causes considerable damage to cultivated fruits trees, particularly stone fruits and apples in the temperate regions of the World and during the postharvest with an important economic impact. In particular in Italy, it is important to monitor the Monilinia population to control economic losses associated to the peach and nectarine market. This motivates the research steps presented in this dissertation on Monilinia Italian isolates. The Monilinia species collected from stone fruits have been identified using molecular analysis based on specific primers. The relevant role of M. fructicola was confirmed and, for the first time, it was found also on apple fruits. To avoid the development of resistant strains and implement valid treatment strategies, the understanding of the fruit natural resistance during different developmental stages and the assessment of the Monilinia sensitivity/resistance to fungicides are required. The relationship between the inhibition spots and the phenolic compounds in peach fruit peel was highlighted in this research. Three methods were used to assess isolate resistance/sensitivity, the amended medium, the Spiral Gradient Endpoint Method (SGD) and the Alamar Blue method. The PCR was used to find possible mutation points in the b-tubulin gene that is responsible for fungicide resistance. Interestingly, no mutation points were observed in resistant M. laxa isolates, suggesting that the resistance could be stimulated by environmental factors. This lead to the study of the effect of the temperature on the resistance and the preliminary results of in vitro tests showed that maximum inhibition was observed at 30°C.

Use of Biotechnology to increase the content of bioactive compounds in fermented foods of plant origin

The objectives of this PhD research were: i) to evaluate the use of bread making process to increase the content of β-glucans, resistant starch, fructans, dietary fibers and phenolic compounds of kamut khorasan and wheat breads made with flours obtained from kernels at different maturation stage (at milky stage and fully ripe) and ii) to study the impact of whole grains consumption in the human gut. The fermentation and the stages of kernel development or maturation had a great impact on the amount of resistant starch, fructans and β-glucans as well as their interactions resulted highly statistically significant. The amount of fructans was high in kamut bread (2.1g/100g) at the fully ripe stage compared to wheat during industrial fermentation (baker’s yeast). The sourdough increases the content of polyphenols more than industrial fermentation especially in bread made by flour at milky stage. From the analysis of volatile compounds it resulted that the sensors of electronic nose perceived more aromatic compound in kamut products, as well as the SPME-GC-MS, thus we can assume that kamut is more aromatic than wheat, so using it in sourdough process can be a successful approach to improve the bread taste and flavor. The determination of whole grain biormakers such as alkylresorcinols and others using FIE-MS AND GC-tof-MS is a valuable alternative for further metabolic investigations. The decrease of N-acetyl-glucosamine and 3-methyl-hexanedioic acid in kamut faecal samples suggests that kamut can have a role in modulating mucus production/degradation or even gut inflammation. This work gives a new approach to the innovation strategies in bakery functional foods, that can help to choose the right or best combination between stages of kernel maturation-fermentation process and baking temperature.

Study of the effect of minor compounds on the oxidative stability and physical properties of bulk oil, oil-in-water emulsion, and food emulsions

Minor components are of particular interest due to their antioxidant and biological properties. Various classes of lipophilic minor components (plant sterols (PS) and α-tocopherol) were selected as they are widely used in the food industry. A Fast GC-MS method for PS analysis in functional dairy products was set up. The analytical performance and significant reduction of the analysis time and consumables, demonstrated that Fast GC-MS could be suitable for the PS analysis in functional dairy products. Due to their chemical structure, PS can undergo oxidation, which could be greatly impacted by matrix nature/composition and thermal treatments. The oxidative stability of PS during microwave heating was evaluated. Two different model systems (PS alone and in combination) were heated up to 30 min at 1000 W. PS degraded faster when they were alone than in presence of TAG. The extent of PS degradation depends on both heating time and the surrounding medium, which can impact the quality and safety of the food product destined to microwave heating/cooking. Many minor lipid components are included in emulsion systems and can affect the rate of lipid oxidation. The oxidative stability of oil-in-water (O/W) emulsions containing PS esters, ω-3 FA and phenolic compounds, were evaluated after a 14-day storage at room temperature. Due to their surface active character, PS could be particularly prone to oxidation when they are incorporated in emulsions, as they are more exposed to water-soluble prooxidants. Finally, some minor lipophilic components may increase oxidative stability of food systems due to their antioxidant activity. á-tocopherol partitioning and antioxidant activity was determined in the presence of excess SDS in stripped soybean O/W emulsions. Results showed that surfactant micelles could play a key role as an antioxidant carrier, by potentially increasing the accessibility of hydrophobic antioxidant to the interface.

Actividad antioxidante de los compuestos fenólicos contenidos en las semillas de la vid (vitis vinifera l.)

El propósito de este trabajo es obtener un antioxidante natural a partir de las semillas de uva (Vitis vinifera L.), para emplear en alimentos. Para ello se compararon distintos solventes para la extracción de fenoles de las semillas de la uva, de modo de obtener el extracto más concentrado en compuestos activos con la mínima degradación de su poder antioxidante durante el proceso de obtención. La concentración de fenoles totales de los extractos se determinó por el método Folin Ciocalteu. El poder reductor de los extractos se midió empleando el método de Oyaizu. Una vez seleccionado el solvente más adecuado para la extracción, se analizó la cinética de extracción, optimizando el tiempo de tratamiento. El extracto fue concentrado al vacío, y se veríficó la conservación del poder reductor en el extracto concentrado, por el método de Oyaizu. El extracto de semillas concentrado y sin concentrar se empleó en un sistema real sujeto a oxidación, tal como el jugo de manzanas. El grado de oxidación del jugo se midió por el método de Özoglu. El extracto concentrado fue deshidratado por secado en lecho de espuma y por liofilizado. En ambos casos se verificó el efecto del tratamiento de secado sobre el poder reductor. Finalmente, se evaluó la actividad antioxidante del extracto líquido concentrado de semillas de vid, respecto de otros antioxidantes comerciales como ácido ascórbico y dióxido de azufre. El sustrato oxidable fue el jugo de manzanas, y el grado de oxidación se midió por el método de Özoglu. El análisis estadístico de los datos se realizó mediante el análisis de la varianza; cuando no fue posible emplear el mencionado análisis, debido a que no se verificaban los supuestos básicos para su aplicación, se empleó la prueba de Kruskal –Wallis. En todos los casos, se utilizó el programa Statgraphics plus ®4.0. Para obtener un extracto antioxidante a partir de semillas de vid se utilizó una ex-tracción con agua a 90ºC, durante 4 horas. La relación sólido- líquido empleada fue de 1g de semillas enteras por 10 ml de solvente. El extracto obtenido presentaba una concentración de 12,587 mg de fenoles totales por gramo de semillas de uva extractadas y un poder reductor de 1,290 unidades. Como consecuencia del análisis de la cinética de extracción, el tiempo de tratamiento se redujo de 4 horas a 3 horas. La concentración del extracto se realizó al vacío a 60ºC, verificándose un aumen-to del poder reductor en el extracto concentrado, comprobado sobre jugo de manzanas. Comparando el extracto concentrado y el extracto sin concentrar se observa que la concentración de fenoles totales aumentó 29,57 veces, mientras que el poder reductor aumentó 37,39 veces. El deshidratado del extracto por medio del lecho de espuma permitió conservar el poder reductor del mismo, no ocurrió lo mismo en el deshidratado por liofilizado, donde se produjo un deterioro del poder reductor. Para un mismo contenido de fenoles totales agregado al jugo de manzanas, el ex-tracto líquido sin concentrar produjo un 28,4% de inhibición de la oxidación, mientras que el de extracto líquido concentrado produjo un 51,5 % de inhibición de la oxidación del jugo de manzanas. El extracto de semillas de vid, aplicado como antioxidante en jugo de manzanas, inhibió el desarrollo de la oxidación en un 31,51%, considerando 24 horas el tiempo de tratamiento. Este desempeño supera al ácido ascórbico, que en iguales condiciones, inhibió el desarrollo de la oxidación en un 2,6%. Pero en las condiciones de tra-bajo, el dióxido de azufre resulta mejor antioxidante que ambos, ya que logró inhibir el desarrollo de la oxidación en un 97,40 %.

Extracto de semillas de vid (Vitis vinifera L.) con actividad antioxidante : eficiencia de diferentes solventes en el proceso de extracción

Los objetivos de este trabajo fueron comparar la eficiencia de diferentes solventes en la extracción de compuestos fenólicos a partir de las semillas de vid; evaluar la actividad antioxidante a través del poder reductor de los extractos obtenidos y analizar el proceso de extracción a diferentes temperaturas, utilizando el solvente más eficiente. Se emplearon semillas de vid cv. Cabernet Sauvignon provenientes de vinificaciones. Los solventes ensayados y las temperaturas de tratamiento fueron los siguientes: agua destilada a 90°C, alcohol metílico al 70% a 30°C, acetona al 75% a 30°C y alcohol etílico al 20% a 30°C. La concentración de los compuestos fenólicos fue determinada por el método de Folin-Ciocalteu. La actividad antioxidante se determinó midiendo el poder reductor, por el método de Oyaizu. El agua a 90°C fue el solvente más eficiente para la extracción de compuestos fenólicos de las semillas de la vid (12,587 mg/g de materia seca). La acetona 75% a 30°C extrajo 7,268 mg de compuestos fenólicos / g de materia seca, el metanol al 70% a 30°C extrajo 0,963 mg de compuestos fenólicos / g de materia seca y el etanol al 20% a 30°C extrajo 0,799 mg de compuestos fenólicos / g de materia seca. Se estudió el proceso de extracción empleando agua a 60 y a 90°C. El agua a 90°C extrajo aproximadamente cinco veces más polifenoles que el agua a 60°C, después de cinco horas de tratamiento.

Extracto de semillas de vid (Vitis vinifera L.) con actividad antioxidante : concentración, deshidratación y comparación con antioxidantes de uso comercial

Los objetivos de este trabajo fueron: 1) Evaluar un proceso de concentración y deshidratación de un extracto polifenólico de semillas de vid (Vitis vinifera L.). El proceso debía conservar el poder reductor del extracto. 2) Valorar la eficiencia del antioxidante obtenido por comparación con antioxidantes de uso habitual en un producto vegetal susceptible de ser oxidado, tal como el jugo de manzanas. El extracto de semillas de vid fue concentrado a 60°C en un concentrador rotativo al vacío de laboratorio. La concentración de los compuestos fenólicos fue determinada por el método de Folin-Ciocalteu modificado. La actividad antioxidante se determinó midiendo el poder reductor, por el método de Oyaizu. La oxidación del jugo de manzanas se determinó por el método de Özoglu. El extracto concentrado obtenido fue deshidratado por dos métodos: liofilización y secado en lecho de espuma. A igual concentración fenólica, el extracto concentrado mostró mayor capacidad antioxidante que el extracto simple. El extracto secado en lecho de espuma conservó la misma capacidad antioxidante que el extracto concentrado. El extracto liofilizado experimentó una pérdida significativa de la actividad antioxidante. El extracto concentrado de semillas de vid inhibió la oxidación del jugo de manzanas en un 31,51%, el ácido ascórbico en un 2,60% y el dióxido de azufre en un 97,40%.

Polpa de guavira (Campomanesia cambessedeana Berg) desidratada em spray dryer: efeitos das condições de processo e composição da alimentação nas propriedades físico químicas e atividade antioxidante

O objetivo desse trabalho foi obter polpa de guavira desidratada por atomização, utilizando maltodextrina ou goma arábica como agentes carreadores. Inicialmente, avaliou-se a influência das condições de processo, temperatura do ar de secagem (130, 155 e 180) °C e vazão volumétrica da mistura (20 e 40) mL/min, o tipo e concentração de agente carreador (10 e 20) % nas características físicas, físico-químicas e atividade antioxidante do produto obtido. As propriedades analisadas foram umidade, atividade de água, higroscopicidade, solubilidade, cor, distribuição e tamanho médio de partículas, morfologia, compostos fenólicos totais e atividade antioxidante. A temperatura do ar de secagem e a vazão volumétrica de alimentação influenciaram significativamente todas as propriedades da guavira em pó. A umidade e atividade de água apresentaram os menores valores na temperatura intermediária, independentemente do tipo e concentração do carreador usado. A solubilidade das amostras adicionadas de maltodextrina foram superiores às amostras com goma arábica. O aumento da concentração de agente carreador, em geral, proporcionou um aumento no parâmetro L* e diminuição dos parâmetros a* e b*, tornando as amostras mais claras e reduzindo as tonalidades vermelha e amarela. A guavira em pó apresentou coloração próxima do amarelo e marrom, com grande variação nos parâmetros de cor C* e H* em função das diferentes condições de secagem. A distribuição do tamanho de partículas não teve um padrão definido e o tamanho médio das amostras com maltodextrina foram maiores do que as com goma arábica para a temperatura do ar a 130 °C. No entanto, para as outras temperaturas (155 e 180) °C não houve um comportamento específico do tamanho das partículas em função da vazão de alimentação, tipo e ou concentração de agente carreador. A análise de microscopia eletrônica de varredura permitiu observar que as partículas obtidas tanto com maltodextrina como goma arábica apresentaram formato esférico, superfície rugosa e com adesão de partículas menores nas de maior tamanho, sendo que a superfície das partículas com goma arábica também apresentaram concavidades. A atividade antioxidante foi superior quando utilizada a temperatura de secagem intermediária. A partir das condições selecionadas na primeira etapa (temperatura do ar de 155 °C, vazão volumétrica da mistura de 40 mL/min e 10% de maltodextrina ou goma arábica) a polpa de guavira em pó foi caracterizada quanto a temperatura de transição vítrea, as isotermas de adsorção e a estabilidade à estocagem do ácido ascórbico, compostos fenólicos totais e da atividade antioxidante da polpa de guavira em pó produzida por spray drying ao longo de 120 dias. As temperaturas de transição vítrea foram de (25,2 ± 2,7 °C e 31,4 ± 0,4) °C para os pós produzidos com goma arábica e maltodextrina, respectivamente. O modelo de BET apresentou ajuste muito bom (R2>0,99) para descrever o comportamento de sorção de água das amostras nas temperaturas de (20, 30 e 40) °C. A polpa de guavira em pó produzida com goma arábica apresentou maior adsorção de água do que as amostras obtidas com maltodextrina. No estudo da estabilidade, as amostras foram acondicionadas em embalagem de polietileno laminado e armazenadas a 25 °C e umidade relativa de 75%. A embalagem de polietileno laminado foi eficiente na manutenção do teor de ácido ascórbico e atividade antioxidante da guavira em pó por um período de 120 dias, independente do carreador adicionado. O teor de compostos fenólicos para a guavira em pó com goma arábica apresentou uma redução nos primeiros 22 dias, contudo a amostra com maltodextrina manteve-se estável durante 120 dias de armazenamento.

Filmes e coberturas a base do resíduo da extração de corante de cúrcuma

O objetivo deste trabalho foi avaliar o potencial de uso do resíduo da extração de pigmento de cúrcuma na produção de filmes e coberturas. Para o estudo dos filmes, foram utilizados glicerol e sorbitol como plastificantes e avaliados os efeitos da concentração de farinha de cúrcuma e do plastificante sobre as propriedades mecânicas, solubilidade, permeabilidade ao vapor de água (PVA), molhabilidade, atividade antioxidante, teor de curcuminóides e teor de compostos fenólicos totais utilizando um Delineamento Central Composto Rotacional 22, e os resultados foram avaliados utilizando a metodologia de superfície de resposta (MSR). A concentração de farinha afetou de forma positiva a espessura, PVA e o teor de curcuminóides totais dos filmes plastificados com glicerol e sorbitol. Entretanto, esta variável afetou as propriedades de solubilidade, molhabilidade e teor de compostos fenólicos totais somente dos filmes com glicerol. A concentração de plastificante (glicerol ou sorbitol) afetou significativamente a solubilidade, PVA e molhabilidade de ambos os filmes. Filmes de farinha de cúrcuma com boas propriedades mecânicas, baixa permeabilidade ao vapor de água, alta atividade antioxidante, alto teor de curcuminóides e alto teor de compostos fenólicos totais podem ser produzidos utilizando 27,9 a 30 g glicerol/100 g farinha ou 30 a 42 g sorbitol/100 g farinha e concentração de farinha na faixa de 5% a 6,41%. A cobertura de farinha de cúrcuma contendo 6% de farinha e 30 g glicerol/100 g de farinha foi aplicada em bananas Maçã (Musa acuminata) armazenadas a 27ºC e 65% UR. Assim, foi avaliado o efeito da cobertura na qualidade pós-colheita das bananas em função à suas características físico-químicas como perda de massa, firmeza da polpa, pH, acidez titulável, sólidos solúveis, açúcares redutores e cor da casca. Os resultados mostraram que a cobertura foi eficiente em diminuir a perda de massa, o teor de açúcares redutores, a acidez, a perda da firmeza e a cor da casca principalmente durante a etapa de maturação do fruto. Entretanto, não foi observado grande efeito da cobertura sobre o pH e o teor de sólidos solúveis durante o período estudado. As bananas sem a cobertura tiveram vida útil de 6 dias, enquanto as bananas com cobertura tiveram vida útil de 9 dias.

Bioactive compounds and physical and chemical characteristics of mini tomatoes: influence of postharvest treatments

Tomatoes are among the most cultivated and used vegetables in the world. They are very succeptible to post harvest losses due to high perishability, therefore the use of postharvest treatments may contribute to conservation of this fruit, however the treatments might affect significantly physico-chemical, sensory and nutritional characteristics of tomatoes. Given the perishability of tomato and the economic importance of small tomato fruits, the purpose of the present study was to determine the effect of gamma radiation, carnauba coating and 1-MCP treatments on tomato fruit quality during storage. The study may be divided into two parts. In the first, mini tomatoes cv. Sweet Grape were harvested at breaker stage, divided into 4 grous and treated with gamma radiation (0.6 kGy), carnauba coating (1 L 1000 kg-1) and 1-MCP (500 nL L-1) and then stored at 25±2°C for 30 days with a control group of tomatoes. In the seconnd part, tomatoes harvested at light-red stage were submitted to the same treatments and storage period. Every 6 days tomatoes were evaluated for color modifications, fruit firmness, souble and total pectin (only for light-red tomatoes), mass loss, titratable acidity (TA), soluble solids (SS), SS/TA ratio, carotenoids profile, formation of lycopene isomers, total phenolic compounds, ascorbic acid and antioxidant capacity. For tomatoes harvested at breaker stage and submitted to the treatments the results showed mass loss was delaying mainly by carnauba wax, and to a lesser extend by 1-MCP. Fruit firmness were better retained for 1-MCP treated fruits and carnauba treatment showed a transient effect in preserving fruit firmness. SS/TA of tomatoes treated with gamma radiation and carnauba presented no differences from control values, and were lower with the application of 1-MCP. Color was negatively affected by 1-MCP and earlier changed (6th day) when gamma radiation was applied. In relation to bioactive compounds of tomatoes harvest at breaker stage, results indicated gamma radiation and 1-MCP decreased the final content of lycopene and produced more (Z)-isomers of lycopene. Gamma radiation also induced a decreased in ?-carotene and an increased in phenolic compounds by the end of storage period. 1-MCP treatment promoted a slow down increase in ascorbic acid content during storage. Antioxidant capacity of the hydrophilic fraction was not dramatically affected by treatments and the lipophilic fraction was lower, especially for 1-MCP fruits. In addition, contents of ?-carotene, lycopene, (Z)-isomers of lycopene, ascorbic acid and antioxidant capacity increased during the period of storage while contents of lutein and phenolic compounds tended to decrease. Regarding tomatoes harvest at light-red stage, the most effective treatments for delaying fruit firmness and mass loss was carnauba and 1-MCP, while gamma radiation was the treatment with higher mass loss and the less fruit firmness, which could be associated with the higher solubilization of pectins promoted by radiation treatment. Color (L* and Hue) was mainly affected by 1-MCP treatment which delayed color development, however, by the end of storage, the values were not different from the other treatments. SS/TA ratio was lower for fruits treated with 1-MCP and TA was not so dramatically affected by treatments. Furthermore, mini tomatoes harvested at light-red stage, demonstrated irradiation induced changes in the final content of lycopene, increasing it, and formed less (13Z)-lycopene, while 1-MCP and carnauba coating slow down the increase in lycopene and slown down the decrease of ascorbic acid and phenolic compounds. Antioxidant capacity of lipophilic fraction was not affected by treatments and the hydrophilic fraction was lower for irradiated fruits only on day 0 as well as phenolic compounds. In the other days, no differences among treatments were observed for hydrophilic antioxidant capacity. Considering the results, the best combination of SS and TA and fruit preservation for mini tomatoes harvest at breaker stage was promoted by carnauba coating, which seems to be the treatment that causes fewer changes in bioactive compounds of breaker tomatoes. However, when mini tomatoes were harvested at light-red stage, SS/TA ratio and color were better and, to preserve the quality of these fruits, besides carnauba coating, 1-MCP also could be indicated

Degradação anaeróbia de fenol sob diferentes condições nutricionais

A tecnologia anaeróbia tem sido utilizada com sucesso no tratamento de água residuária contendo compostos fenólicos. Recentes pesquisas incluem tais compostos entre aqueles que podem ser degradados através desse processo. O objetivo desse trabalho foi avaliar a degradação do fenol em diferentes condições nutricionais, com ênfase na redução do sulfato. Os experimentos foram realizados com meio de cultura específico para esses microrganismos anaeróbios. Foram realizados ensaios de degradação em reatores em batelada alimentados nas seguintes condições: (1) fenol e sulfato, a diferentes concentrações, com inóculo previamente enriquecido; (2) fenol, sulfato e co-substratos e; (3) fenol, sulfato e extrato de levedura. Todos os ensaios foram realizados em temperatura de 30 graus Celsius, sob agitação de 150 rpm. Foi avaliado o consumo de fenol e sulfato e, produção de metano, em função do tempo, para diferentes concentrações iniciais de fenol e sulfato. Nos ensaios com reatores alimentados com fenol (329,3 mg/l); fenol (307,3 mg/l) e sulfato (160 mg/l); fenol (322.3 mg/l), sulfato (160 mg/l) e lactato (478,16 mg/l); fenol (332,1 mg/l), sulfato (150 mg/l) e etanol (129,76 mg/l), a remoção foi de, respectivamente, 99,8%, 98,2%, 98,8% e 98,8%. Os reatores alimentados com fenol (239,7 mg/l) obtiveram 100% de eficiência na degradação em apenas 11 dias e, os reatores alimentados com fenol (234,3 mg/l) e sulfato (162,5 mg/l) e fenol (256,0 mg/l) e sulfato (500 mg/l) tiveram eficiências de degradação de, respectivamente, 98,8% e 99,3% com 17 dias de operação. Tais eficiências foram obtidas pelo acréscimo de extrato de levedura nos reatores, no início dos ensaios. A caracterização morfológica foi realizada através de microscopia óptica. A diversidade microbiana referente aos Domínios Bacteria e Archaea, além do grupo de bactérias redutoras de sulfato foi avaliada através da técnica de PCR DGGE, onde foram observadas alterações nas populações microbianas, em função das condições nutricionais. Para o Domínio Archaea não foram observadas diferenças nos ensaios realizados. Para o Domínio Bacteria e Grupo das BRS essas diferenças foram, mais facilmente, percebidas com relação ao inóculo e entre os diversos reatores. A alteração na diversidade microbiana pode ter sido decorrente da composição do meio que, nesse caso, foi específico para BRS e a composição do inóculo que continha parte previamente adaptada às BRS. Essas condições adequadas puderam propiciar surgimento e desenvolvimento de populações microbianas capazes de degradar fenol, utilizando sulfato.

Extração, determinação da composição fenólica e avaliação do potencial de desativação de espécies reativas de oxigênio e da atividade anti-inflamatória de resíduos de amendoim, pimenta-rosa e pimenta-do-reino

O setor agroindustrial tem se expandido muito nos últimos anos, levando o país a um aumento na geração de resíduos agroindustriais, sendo que a maior parte deles ainda é descartada no meio ambiente, sem tratamento adequado, ou utilizada na alimentação animal, destinos que a priori, não geram ganhos econômicos para a agroindústria além de representarem gargalos logísticos e ambientais na sua disposição. Nesse sentido, o presente trabalho teve como objetivos otimizar o processo de extração de compostos bioativos, avaliar in vitro as atividades antioxidante, por meio da desativação de espécies reativas de oxigênio, e anti-inflamatória, bem como determinar a composição fenólica dos resíduos agroindustriais a saber: película de amendoim (Arachis hypogaea) (cultivares IAC886 e IAC505), pimenta-rosa (Schinus terebinthifolius Raddi) e pimenta-do-reino (Piper Nigrum L). O processo de otimização da extração de compostos antioxidantes foi realizado utilizando dois processos de extração, extração convencional e subcrítica, em delineamento composto central rotacional, utilizando como variáveis a temperatura e tempo e a pressão e temperatura, respectivamente, com os solventes etanol 80%, água e propilenoglicol 80%. Durante o processo de otimização a atividade antioxidante foi avaliada pelo método de sequestro do radical ABTS. Os melhores resultados foram obtidos para a extração convencional com os solventes etanol 80%, água e propilenoglicol 80%. A película de amendoim (IAC505) apresentou as maiores atividades antioxidantes (1.396,67, 580,44 e 859,89 μmol.g-1 em equivalentes de trolox, para os solventes etanol 80%, água e propilenoglicol 80%, respectivamente). A partir dos resultados obtidos para os solventes testados, utilizando a extração convencional, foram feitas outras análises de atividade antioxidante considerando o tempo e temperatura ideal de extração. Foram realizadas análises de avaliação da capacidade de redução de Folin-Ciocalteau e potencial de desativação dos radicais livres sintéticos (DPPH e ABTS) e espécies reativas de oxigênio (radicais peroxila, superóxido e ácido hipocloroso). O solvente de extração que apresentou melhores resultados em todos os ensaios foi o etanol 80%, sendo, portanto o solvente utilizado nas análises subsequentes. A partir da definição do melhor sistema extrator foram realizadas análises da composição fenólica, por meio das técnicas de cromatografia líquida de alta eficiência em modo analítico (HPLC-RP), cromatografia gasosa acoplada com espectrometria de massas (GC-MS), e avaliação in vitro da atividade anti-inflamatória. Foram identificados nos resíduos estudados procianidinas B1 e B2, ácido p-cumárico, miricetina, ácido ferúlico, ácido siríngico, ácido sinápico, epicatequina e catequina. A pimenta-do-reino diminui significativamente os níveis de TNF-α e nitritos, reduzindo assim o processo inflamatório gerado. Os resultados obtidos neste trabalho demonstram que estes resíduos agroindústrias possuem grande potencial biológico, podendo assim ser melhores aproveitados tanto pela indústria de alimentos quanto pela indústria farmacêutica.