Diploma thesis deals with the characterization of chokeberry (Aronia melanocarpa) and saskatoon berry (Amelanchier alnifolia) fruits. The theoretical part focuses on the botanical classification, chemical composition, curative effects and the use of chokeberry and saskatoon berry in the food industry. Further, the attention is also devoted to a review of methods for the determination of important antioxidants, specialized on the determination of polyphenols, anthocyanins, ascorbic acid and total antioxidant activity. The experimental part of the work deals with the determination of selected parameters in different varieties of chokeberry and serviceberry, harvested in 2012 and 2013 using spectroscopic and chromatographic techniques with an emphasis on effect of solvent (50% ethanol, destilled water, DMSO). Specifically total content of polyphenols and anthocyanins, color characteristics using the CIE L*a*b* system and the ability of the sample to reduce the iron are determined using the UV-VIS-NIR spectroscopy. Further ascorbic acid concentration in individual extracts is quantified by High performance liquid chromatography coupled with a diode array detector and the total antioxidant activity is characterized by electron paramagnetic resonance involving •DPPH and ABTS•+ assays.

Ovocné šťávy jsou celosvětově nejoblíbenějšími nealkoholickými nápoji. Kromě vitamínů a minerálů jsou také bohatým zdrojem polyfenolů, které jsou považovány za látky přispívající k celkové antioxidační aktivitě. Zvyšující se požadavky konzumentů na kvalitu a bezpečnost potravin nutí výrobce k zvyšování výrobních a kvalitativních standardů potravin. Nedávné inovace ve výrobě ovocných šťáv vedly k modifikacím výrobních podmínek, které zahrnují změny ve složení šťávy (např. přídavek ovocné dužiny nebo malých ovocných kousků), vylepšování vlastností obalových materiálů a modifikace produkční atmosféry. Cílem diplomové práce bylo pomocí metod EPR a UV-VIS spektroskopie posoudit vliv různých výrobních postupů na vybrané kvalitativní znaky (antioxidační aktivita, změna barvy a koncentrace vybraných fytochemikálií) pomerančových šťáv s dužinou a ananasových šťáv, a charakterizovat jejich změny v průběhu dlouhodobého skladování. Získané výsledky byly zpracovány metodami multivariační statistické analýzy s cílem posoudit vliv původu suroviny, roku produkce a použitých výrobních podmínek na sledované parametry a celkovou kvalitu ovocných džusů. Výsledky jasně prokázaly, že pasterizace ovlivňuje kvalitu výrobku; ačkoli byla použita „šetrná“ pasterizace, byl u těchto vzorků pozorován mírný pokles prakticky všech monitorovaných parametrů v porovnání s čerstvými – nepasterizovanými šťávami. Avšak pokud jde o po-pasterizační změny, byl prokázán významný vliv produkční atmosféry na monitorované kvalitativní parametry šťáv, prokazující, že náhrada kyslíku inertními plyny může výrazně prodloužit trvanlivost šťáv. Kinetické studie změn sledovaných parametrů na době skladování ukazují, že ve většině případů dochází k postupnému zhoršování všech monitorovaných parametrů, a to bez ohledu na technologii zpracování šťávy. Ve vzorcích vyrobených použitím modifikované atmosféry bylo, v závislosti na typu inertního plynu, pozorováno zpomalení poklesu jednotlivých kvalitativních parametrů šťáv jako výsledek aplikace inertní atmosféry. Tento trend byl zřejmý především pro delší skladovací období, zatímco na začátku skladování (6 - 7 týdnů po výrobě dané šťávy) byly pozorované trendy nejednoznačné. Lze proto předpokládat že tento „ochranný“ efekt inertní atmosféry je ovlivňován fyzikálně-chemickými vlastnostmi jednotlivých plynů, zejména jejich rozpustností v kapalném médiu. Výsledky také prokázaly že vlastnosti šťáv jsou ,bez ohledu na druh ovoce, výrazně ovlivňovány také sezónními vlivy a původem surové šťávy. Z výsledků statistické analýzy vyplývá, že rok výroby je silným diskriminačním faktorem. Na základě všech monitorovaných charakteristik, bylo dosaženo absolutní diskriminace vzorků pomocí kanonické diskriminační analýzy. Pokud jde o původ surovin, byla dosažena pouze částečná diferenciace, i když s poměrně vysokým klasifikačním skóre. Pokud jde o technologii výroby, byla dosažena pouze částečná diferenciace vzorků podle typu použitého inertního plynu. Získané poznatky pomohou výrobcům ovocných šťáv s optimalizací výrobních podmínek s cílem získat výrobek s maximem prospěšných vlastností, které by si udržel po celou dobu expirace, jejíž délka může být také optimalizovány na základě prezentovaných výsledků.

Antioxidant properties and some other characteristics of selected organic and conventional products (wine, spelt flours) were evaluated in view of their potential utilization as markers suitable for food unambiguous differentiation according to the affiliation to production system. Beside these, study of fruit juices stability during storage in dependence on different technological procedures of juice processing was performed. In both cases, combination of dominantly EPR and UV/VIS spectroscopy but also other methods (e.g., HPLC, AAS, electrophoresis) was applied to evaluate the stability and antioxidant properties of selected samples. Statistical methods, especially methods of multivariate statistics were used to assess the influence of production and technological-processing operations on antioxidant status of foods and their selected characteristics. Qualitative analysis of Slovak and foreign organic and conventional wines proved that their antioxidant properties are influenced by production system, vintage and variety. On the basis of statistical analysis, descriptors of AAE, kTEMPOL and %RS were selected as the most powerful markers for Slovak organic and conventional wines discrimination. The discrimination of white and red organic and conventional wines possessed 100% and 95.2% correctness, respectively. High correctness of differentiation of Slovak and foreign wines was also achieved. Monitored descriptors of antioxidant activity of spelt flours varied in dependence on season, origin, variety, production system and depend also on way of spelt treatment, although this effect is not so significant. It was also confirmed that the type of extraction agent substantially affects the composition of extracts including polyphenols and flavonoids concentration, affecting thus also their antioxidant activity. Mixture of 50% ethanol/water was selected as the most promising extraction system. Although the results of individual spectroscopic assays revealed high variability and often ambiguous trends, the differences found were sufficient for successful differentiation of both wines and spelt flour samples according to the selected criteria by multivariate statistical processing of the data. Influence of changes in technological procedures of fruit juice processing (nitrogen atmosphere application, utilization of caps with oxygen scavengers) on radical-scavenging properties of 100% pineapples juices with pieces of pineapple and 100% orange juices with pulp was assessed as well. Results obtained proved that antioxidant activity of juices varied in dependence on storage conditions (temperature, storage time, light exposure) and depends also on origin, variety and quality of fruit used. However, the application of nitrogen atmosphere and active packaging materials is not sufficient enough to eliminate the oxidation reactions in juices, but changes in antioxidant activity are better predictable. It can be concluded that EPR spectroscopy could be effectively applied for wines and spelt flours differentiation according to the way of production and to assess the stability of food products, either alone or in combination of UV/VIS spectroscopy and the other methods, utilizing multivariate statistics for processing of experimental data.

Present diploma thesis is focused on natural antimicrobial substances. The aim of the experimental part of thesis was testing of antimicrobial activity of selected natural substances in free and encapsulated forms, preparation of particles and monitoring of their long-term stability. Various teas extracts (aqueous, ethanoic and DMSO) and aqueous herbs extracts were used as plant antimicrobial substances. Liposomes and chitosan particles with encapsulated aqueous teas extracts were prepared by ultrasonication. Encapsulation efficiency, size, stability and antioxidant activity for all particles were specified. Consequently, within a two months period, long-term stability for these particles was monitored. Spectrophotometric method for determining total polyphenols was used for evaluating encapsulation efficiency as well as for monitoring of long-term stability. The DLS method was used for determination of size and stability of particles. Antioxidation activity was monitored by using spectrophotometric method using ABTS. For testing antimicrobial activity of various types of teas, herbs extracts and their mixtures and combianitons with lysozyme and antibiotics, broth dilution methods were used while using both Gram positive and Gram negative bacterial test strains. Antimicrobial activity of prepared particles was also monitored, the antimicrobial activity was especially significant with the chitosan particles. Suitable combinations of natural substances whatever with antimicrobial or antioxidation effect either in free or encapsulated form could be used for applications for example in cosmetics industry, food processing or in medicine.

MSc (Botany) / Department of Botany / See the attached abstract below

Orientador: Marcelo de Almeida Silva / Resumo: A soja possui grande importância no mercado mundial, e para atender a sua crescente demanda é preciso aumentar a produtividade, que tem se tornado um desafio em condições de ausência de chuva. Para contornar os efeitos negativos do déficit hídrico, tem-se adotado práticas agrícolas com a aplicação de bioestimulantes como os compostos por ácidos fúlvicos e extrato da alga marinha Ascophyllum nodosum (L.), que atuam na melhoria do desenvolvimento das plantas em condições estressantes. Neste estudo, objetivou-se compreender como ácidos fúlvicos e extrato da alga marinha Ascophyllum nodosum (L.) atuam em conjunto nos mecanismos fisiológicos e bioquímicos das plantas e possibilitando a mitigação dos efeitos negativos gerados pelo estresse, e ainda determinar qual formulação e dose do bioestimulante tem melhor efeito sobre o desenvolvimento de plantas de soja submetidas ao déficit hídrico. O experimento foi conduzido em casa de vegetação, em vasos contendo 10 kg de solo sob delineamento experimental de blocos casualizados em fatorial duplo com dois fatores adicionais, com cinco repetições e 11 tratamentos constituídos por três formulações do bioestimulante e três doses (0,25; 0,50 e 1,0 kg ha-1), controle com déficit hídrico e controle sem déficit hídrico. As plantas foram submetidas a 50% da capacidade de campo por três dias, em seguida foram reidratadas e receberam a aplicação do bioestimulante, com exceção dos controles. Foram avaliados parâmetros de crescimento, fisiológicos, b... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Soy has great importance in the world market, and to meet its growing demand it is necessary to increase productivity, which has become a challenge in conditions of absence of rain. To circumvent the negative effects of the water deficit, agricultural practices have been adopted with the application of biostimulants such as those composed of fulvic acids and extract from the seaweed Ascophyllum nodosum (L.), which act in improving the development of plants in stressful conditions. In this study, the objective was to understand how fulvic acids and extract of the seaweed Ascophyllum nodosum (L.) act together in the physiological and biochemical mechanisms of plants and enabling the mitigation of the negative effects generated by stress, and also determine which formulation and dose of biostimulant has a better effect on the development of soybean plants submitted to water deficit. The experiment was carried out in a greenhouse, in pots containing 10 kg of soil under a randomized double-block design with two additional factors, with five replications and 11 treatments consisting of three biostimulant formulations and three doses (0.25; 0.50 and 1.0 kg ha-1), control with water deficit and control without water deficit. The plants were submitted to 50% of the field capacity for three days, then they were rehydrated and received the application of the biostimulant, with the exception of the controls. Growth, physiological, biochemical parameters and productivity components were e... (Complete abstract click electronic access below) / Doutor

This Ph.D. thesis deals with the deep analysis of polyphenols effects toward vascular disorders. This work provides a number of experimental results of studying both the effects of natural bioactive compounds in red wines and potatoes, and their application to the experiments which includes experimental animal models with spontaneous hypertension. Methods and equipment used in experimental studies allowed us to make several new statements regarding the universal nature of the relationships between the antioxidant capacity and the polyphenolic content in examined foodstuffs. Besides this it is also recognized that food is a source of minerals which also contribute to the total antioxidant capacity and therefore may have influence the endogenous antioxidant enzyme system by providing the essential cofactors. Experimentally we have found that the concentrations of magnesium known for its therapeutic action in wine and red wine extract are comparable to the mineral waters recommended for the treatment and prevention of cardiovascular and metabolic diseases. Moreover, synergistic interactions between selected minerals have been found. The key issue of bioavailability of polyphenols for supporting the idea of the beneficial effects of diet rich in fruits and vegetables toward vascular disease prevention...

Diploma thesis deals with determination of individual chemical parameters contained in modern varietes of gooseberries (Grossularia uva-crispa). The theoretical part is focused on the botanical characteristics of the plant, a description of selected varietes, chemical composition and food utilization fruits. The second half of the theoretical part is devoted to the characterization of selected nutritional parameters and methods for determination of total anthocyanins, phenolic compounds, vitamin C and antioxidant activity. Total of 17 varietes of gooseberries, which come from growers Vondráček and the Research and Breeding Institute of Pomology Holovousy Ltd. were analyzed in the experimental part. pH differential method was used to determination total monomeric anthocyanins. Total content of polyphenols were determined by using the Folin-Ciocalteu reagent. Next was determined content of vitamin C in all varieties by HPLC and was evaluated the total antioxidant aktivity using radical DPPH. In conclusion, individual varieties of gooseberries were compared by the contents of the determined chemical parameters.

The biochemical factors affecting post-harvest preservation in apples indicated that well-preserved varieties of apples had increased superoxide dismutase (SOD) activity initially and the activity declined during later storage as apples deteriorated. The SOD link to better preservation correlated with higher phenolic content and free-radical scavenging linked antioxidant activity. Well-preserved varieties were able to maintain a more stable pentose phosphate pathway (PPP) (measured by the activity of glucose-6-phosphate dehydrogenase, G6PDH) throughout the storage period. Proline content increased in all varieties with an increase in proline dehydrogenase (PDH) activity in the initial period indicating proline catabolism supporting potential ATP synthesis. During later storage succinate dehydrogenase (SDH) activity increased while PDH activity declined indicating a shift to tricarboxylic acid cycle and likely NADH generation for ATP synthesis. This shift coupled with the declining SOD activity coincides with rapid deterioration. The guaiacol peroxidase activity (GPX) activity generally declined in late stages indicating post-harvest deterioration. Increasing number of studies have shown that regular intake of fruits and vegetables have clear links to reduced risk of chronic diseases like diabetes and cardiovascular disease. The beneficial effects in many cases have been attributed to the phenolic and antioxidant content of the fruits and vegetables. Apples are a major source of fiber and contain good dietary phenolics with antioxidant function. Previous epidemiological studies have indicated that intake of apples reduces the risk of developing Type II diabetes. Our studies indicate that this reduced risk is potentially due to modulation of postprandial glucose increase by phenolics present in apples via inhibition of a-glucosidase. Phenolic content was evaluated during 3 months of post-harvest storage of four varieties of apples and results indicated positive linkage to enhanced post-harvest preservation and a-glucosidase inhibition. These in vitro results along with existing epidemiological studies provide strong biochemical rationale for further animal or human clinical studies.

I det här projektet extraherades värdefulla komponenter från äpple-pomace som är en restprodukt från cidertillverkning. Restprodukten var tillhandahållen av en svensk cidertillverkare. Extraktionen fokuserades mot pektin och fenolföreningar som uppvisar antioxiderande aktivitet. Extraktionsprocessen designades ur ett miljövänligt perspektiv, inga skadliga kemikalier användes. Fenolära substanser extraherades initialt från pomacen med en vattenlösning innehållande 50% etanol. Därefter studerades kompositionen och den antioxiderande aktiviteten i dessa extrakt. Fokuseringen i projektet låg främst på pektin. Pektin extraherades med subkritiskt vatten vid tre olika pH (3,5 och 7) samt vid två skilda temperaturer (120°C och 140°C) i antingen 5, 10 och 15 minuters sekvenser. Pektinet karakteriserades med avseende på extraktionsutvinningsgrad, sockerkomposition, molekylvikt och antioxiderande aktivitet. Resultaten jämfördes för att undersöka effekten av pH, temperatur och extraktionstid. Gelningsegenskapen hos olika pektin studerades som ett bevis på koncept i ett empiriskt experiment där hög-viskösa vätskor bestående av 5% pektin och 60% sackaros framställdes. Extraherade fenolföreningar har potential att fungera som naturligt utvunna antioxidanter i kosmetiska produkter och pektin kan tänkas användas som reologimodifierare i vattenbaserade formuleringar med låga pH-värden utan behov av kemiska modifieringar. / In this project, apple pomace from a Swedish cider making factory as a by-product was used as a raw material to extract valuable compounds. The extraction was focused on pectin and phenolic compounds with antioxidant activity. For the extraction procedures environmentally friendly processes were chosen without using any harsh chemicals. Phenolic compounds wereinitially extracted from the pomace using 50% aqueous ethanol and then the composition, total phenolic content and antioxidant activity were studied in these extracts. The main focus was on pectins, which were extracted by subcritical water at three different pH conditions (pH 3, 5 and 7) and two different temperatures (120°C and 140°C) in 5-, 10- and 15 minute sequences. Then the pectins were characterized in terms of extraction yield, sugar composition, molecular weight and antioxidant activity and the results were compared in terms of the effect of pH, temperatur eand extraction time. The gelling properties of the different pectins were also studied as a proof of concept in an empirical experiment, where highly viscous liquids were obtained at 5% pectinand 60% sucrose content. In summary, the extracted phenolic compounds have potential to function as naturally derived antioxidants in cosmetics and the pectin may be used as a rheology modifier in water-based formulations of low pH without any additional chemical modifications.