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This diploma thesis was focused on basic physicochemical properties of caffeine and investigation of the interaction between the polysaccharide sodium hyaluronan (HA), pectin (PEC) or lignohumate (HUM) and the alkaloid caffeine (CAF). The reason why this thesis was focused on study of aqueous solutions of hyaluronan or pectin with caffeine is that the presence of polysaccharide- caffeine complex could be beneficial for slowing the absorption of caffeine. In the theoretical part of the diploma thesis there is described a group of alkaloids, which caffeine belongs to and also polysaccharides hyaluronic acid and pectin. The most attention has been paid to caffeine, its properties and effects on the human organism. This diploma thesis also deals with non-alcoholic beverages with caffeine, 4-methylimidazole, and phosphoric acid. The last chapter of the theoretical part is dedicated to fluorescence spectroscopy. In the experimental part of this work I deal with the basic properties of caffeine and interaction between the caffeine and selected polysaccharides or lignohumate. At first, the chemical and physical properties of caffeine were determined. Based on these results the interaction of caffeine and hyaluronan or pectin was studied by using the fluorescence and absorbance. The HA of molecular weight of 1,7 MDa and pectin from citrus fruits were chosen for the study of the interaction with the caffeine. These two substances were mixed with the caffeine of concentration contained in Coca-Cola. The results of interaction that would lead to the influence of the emission or absorption properties of caffeine, were not proved. As a part of the work there was determined how pectin behaves in strongly acidic solution and then there was monitored the pH, conductivity and solubility of caffeine in water by using thermogravimetry and visual experiments. The obtained results of the interaction of hyaluronan-caffeine, pectin caffeine or caffeine-lignohumate can be used for description of the behavior of caffeine in the presence of selected polysaccharides and natural organic substance.

<p>Stabilizatori na bazi biopolimera postaju posebno povoljna prirodna re&scaron;enja za primenu u kompleksnim sistemima, kao &scaron;to su emulzije tipa ulja u vodi. U ovoj disertaciji je ispitan individualni i kombinovani uticaj tri stabilizatora na bazi polisaharida (vlakna &scaron;ećerne repe, pektin &scaron;ećerne repe i OSA maltodekstrin) na formiranje i stabilnost emulzija kukuruznog ulja u vodi. Ispitan je uticaj četiri različite tehnike emulgovanja (ultrazvučna homogenizacija, visokopritisna homogenizacija, membransko emulgovanje i rotor-stator homogenizacija). Sveobuhvatna karakterizacija koja je obuhvatala ispitivanje fizičko-hemijskih, strukturnih, tenziometrijskih, konduktometrijskih, viskozimetrijskih osobina ispitivanih polisaharida je prikazana u ovoj disertaciji. Dobijeni rezultati veličine kapi emulzija i njihove fizičke stabilnosti ukazuju na dobra stabilizaciona svojstva ispitivanih polisaharida. Interakcije između primenjenih stabilizatora na bazi polisaharida rezultovale su značajnim promenama veličine kapi, zeta potencijala, kriming indeksa i strukture emulzije.</p> / <p>Biopolymer based stabilizers become particularly favorable natural solutions for use in complex systems, such as oil-in-water emulsions. In following dissertation, the individual and combined effects of three polysaccharide based stabilizers (sugar beet fibers, sugar beet pectin and OSA maltodextrin) on the formation and stability of corn oil emulsions in water were tested. The influence of four different emulsion techniques (ultrasonic homogenization, high-pressure homogenization, membrane emulsification and rotor-stator homogenization) were examined. A comprehensive characterization that included the examination of physico-chemical, structural, tensiometric, conductometric, viscometric characteristics of the polysaccharides examined is presented in this dissertation. The obtained results of the emulsion droplet size and their physical stability indicate good stabilization properties of the polysaccharides examined. The interactions between the applied polysaccharide-based stabilizers resulted in significant changes in the droplet size, zeta potential, creaming index and the structure of the emulsion.</p>

Grewia optiva är en utav många outnyttjade stjälkfibrer som skulle kunna bidra till ökandet utav de naturliga fiberalternativen. Fibern har idag inte så många användningsområden på grund utav dess hårda och styva uppbyggnad, vilket gör den svår att spinna till garn. På uppdrag av organisationen Bhartiya Gramotthan Sanstha (BGS) har i detta projekt en redan befintlig metod utvecklats för att förädla fibern. Vad som främst eftersöktes var nedbrytandet av pektin som är en av de faktorer som bidrar till fiberns hårda och styva struktur. I metoden användes biologiskt nedbrytbara enzym som katalysatorer. En fungerande metod skulle kunna öka användningsområdet hos stjälkfibrer generellt och öka möjligheten till användandet utav fler naturliga fibrer. Enzymet som har använts i metoden är ett pektatlyas EC 4.2.2.2 som katalyserar reaktionen som sker då pektinmolekyler klyvs. För att effektivisera processen adderades en komplexbildare, EDTA, som tidigare visat goda resultat för lin. Efter enzymbehandlingen skedde en viktreduktion av fibrerna samt förändring av deras utseende. I svepelektronmikroskop observerades förändring av ytstruktur samt separation mellan fiberbuntarna. Dessa parametrar är viktiga och har stor inverkan på spinnbarheten hos fibrer. I projektet har försök att spinna fibern gjorts men inte lyckats helt. Förändringen på ytstruktur och separation mellan fibrerna tyder dock på att behandlingen är ett steg i rätt riktning. / Grewia optiva is one of many unused bast fibres that could contribute to an increase of natural textile fibres on the industrial market. This fibre has to-day not as many applications due to its stiff and hard structure that makes the fibre difficult to spin into yarn. On behalf of the organisation Bhartiya Gramotthan Sanstha (BGS) has an existing method been developed to process the Grewia optiva fibre. The method is developed to break down substances like pectin that is responsi-ble for the hard and stiff structure of the fibre. Degradable biological en-zymes were used as catalyser in the method. With a functioning method like this the applications of bast fibres could increase and contribute to the use of more natural fibres. The enzyme used to catalyse the chemical reaction and the cleavage of pec-tin molecules in this method was a pectate lyase EC 4.2.2.2. In this method EDTA was used as a chelator to efficient the chemical process. EDTA has been used as a chelator in earlier reports and showed good results. After the enzymatic treatment a weight reduction of the fibre was notable. In SEM-analysis separation between fibres and changes on the fibre surfaces was observed. These parameters are important and affect the spinning capability of the fibre. To test the spinning capability of the enzyme treated fibre they were spun in a ring spinning system, unfortunately not successfully. The surface changes and the separation shows that the enzymatic treatment had occurred and indicates that the method has developed in the right direction.

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Pectic polysaccharides, a class of plant cell wall polymers, form one of the most complex networks known in nature. Despite their complex structure and their importance in plant biology, little is known about the molecular mechanism of their biosynthesis, modification, and turnover, particularly their structure-function relationship. One way to gain insight into pectin metabolism is the identification of mutants with an altered pectin structure. Those were obtained by a recently developed pectinase-based genetic screen. Arabidopsis thaliana seedlings grown in liquid medium containing pectinase solutions exhibited particular phenotypes: they were dwarfed and slightly chlorotic. However, when genetically different A. thaliana seed populations (random T-DNA insertional populations as well as EMS-mutagenized populations and natural variations) were subjected to this treatment, individuals were identified that exhibit a different visible phenotype compared to wild type or other ecotypes and may thus contain a different pectin structure (pec-mutants). After confirming that the altered phenotype occurs only when the pectinase is present, the EMS mutants were subjected to a detailed cell wall analysis with particular emphasis on pectins. This suite of mutants identified in this study is a valuable resource for further analysis on how the pectin network is regulated, synthesized and modified. Flanking sequences of some of the T-DNA lines have pointed toward several interesting genes, one of which is PEC100. This gene encodes a putative sugar transporter gene, which, based on our data, is implicated in rhamnogalacturonan-I synthesis. The subcellular localization of PEC100 was studied by GFP fusion and this protein was found to be localized to the Golgi apparatus, the organelle where pectin biosynthesis occurs. Arabidopsis ecotype C24 was identified as a susceptible one when grown with pectinases in liquid culture and had a different oligogalacturonide mass profile when compared to ecotype Col-0. Pectic oligosaccharides have been postulated to be signal molecules involved in plant pathogen defense mechanisms. Indeed, C24 showed elevated accumulation of reactive oxygen species upon pectinase elicitation and had altered response to the pathogen Alternaria brassicicola in comparison to Col-0. Using a recombinant inbred line population three major QTLs were identified to be responsible for the susceptibility of C24 to pectinases. In a reverse genetic approach members of the qua2 (putative pectin methyltransferase) family were tested for potential target genes that affect pectin methyl-esterification. The list of these genes was determined by in silico study of the pattern of expression and co-expression of all 34 members of this family resulting in 6 candidate genes. For only for one of the 6 analyzed genes a difference in the oligogalacturonide mass profile was observed in the corresponding knock-out lines, confirming the hypothesis that the methyl-esterification pattern of pectin is fine tuned by members of this gene family. This study of pectic polysaccharides through forward and reverse genetic screens gave new insight into how pectin structure is regulated and modified, and how these modifications could influence pectin mediated signalling and pathogenicity. / Pektin Polysaccharide, eine Klasse pflanzlicher Zellwand Polymere, formen eine der komplexesten natürlichen Strukturen. Trotz seiner immensen Bedeutung in der Biologie der Pflanzen sind die Kenntisse über die molekularen Mechanismen der Pektin Biosynthese, dessen Modifikation und Abbau überraschend gering. Eine Möglichkeit neue Einblicke in den pflanzlichen Pektin Metabolismus zu erhalten, ist die Identifizierung von Mutanten mit veränderter Pektinstruktur. Solche Mutanten konnten durch ein neuatiges Selektionsverfahren gefunden werden. Zieht man Keimlinge der Ackerschmalwand (Arabidopsis thaliana) in Flüssigmedium mit Pektinase an, so lässt sich ein typischer Phänotyp beobachten: Die Pflanzen sind kleinwüchsig und leicht chlorotisch. Diesem Verfahren wurden Populationen verschiedener Genotypen (Insertions Linien, EMS Mutanten, natürlich vorkommende Varianten) ausgesetzt. Auf diese Weise wurden Individuen identifiziert, die gegenüber der Pektinase Behandlung eine verminderte oder erhöhte Resistenz aufweisen, was auf eine veränderte Pektinstruktur hindeutet. Die EMS Mutanten wurden einer detaillierten Zellwand Analyse unterzogen. die so in dieser Arbeit identifizierte Kollektion von Mutanten stellt eine wertvolle Ressource für weitere Forschungsansätze zur Regulation, Biosynthese und Modifikation des Pektins dar. Die Lokalisation der Insertionen in den T-DNA Linien führte zur Identifikation interessanter Gene, zu denen der putative Zuckertransporter PEC100 gehört. Dieses Gen steht vermutlich in Verbindung mit der Synthese von Rhamnogalakturonan-I, einem Bestandteil des Pektins. In dieser Arbeit konnte PEC100 im Golgi Apparat, dem Ort der Pektin Biosynthese, lokalisiert werden. Die natürlich vorkommende Variante C24 ist besonders empfindlich gegenüber der Pektinase. Diese Empfindlichkeit konnte anhand rekombinanter Inzucht Linien auf drei bedeutende quantitative Merkmalsloci (QTL) eingegrenzt werden. C24 zeigte zudem ein gegenüber der Referenz verändertes Massenprofil der Oligogalakturonide. Diese werden derzeit als Signalmoleküle in der pflanzlichen Pathogenabwehr diskutiert, was mit der in dieser Arbeit geseigten Resistenz von C24 gegenüber Schwarzfleckigkeit verursachende Pilz (Alternaria brassicicola) korreliert. In einem revers-genetischen Ansatz wurden zudem Mitglieder der Pektin Methyltransferase Familie als potentielle Enzyme getestet, die die Pektin Methylesterifikation beeinflussen könnten. Diese Mutation in einer dieser Methyltransferasen führte zu Veränderungen des Oligogalakturonid Massenprofils. Dies bestätigt die Hypothese, dass Mitglieder dieser Genfamilie an der Regulation der Methylesterifikation von Pektin beteiligt sind. Die vorliegende Studie, in der ein genetishen Selektionverfahren und Methoden der reversen Genetik kombiniert wurden, hat neue Einblicke in die Regulation und Modifikation von Pektin geliefert.

Qualität wird nach ISO 8402 als Gesamtheit von Eigenschaften und Merkmalen eines Produktes, die sich auf deren Eignung zur Erfüllung festgelegter oder vorausgesetzter Erfordernisse beziehen, definiert. Produkte mit hohem Wassergehalt und großer Atmungsrate können erhebliche Qualitätsbeeinträchtigungen in der Nachernteperiode bis zum Verkauf bzw. Verzehr erleiden. Am Beispiel des Spargels, also junger und physiologisch sehr aktiver Pflanzensprosse, werden äußere und innere Qualitätsmerkmale sowie deren Beeinflussbarkeit durch züchterisch- pflanzenbauliche Maßnahmen und Lagerverfahren einschließlich Vorkühlung und Lagerdauer aufgezeigt. Das im Land Brandenburg gewachsene Untersuchungsmaterial wurde aus einem 2- faktoriellen Parzellenfeldversuch, einem Praxis- Sortenvergleichsversuch und durch Zukauf gewonnen. Bonituren und Sortierung nach äußerlich erfassbaren, Qualitätskriterien sowohl von frisch geerntetem als auch von zwei bzw. fünf Wochen unter Kühl- und CA- Lagerbedingungen (verschiedene Gaszusammensetzungen) gelagerten Grün- und Bleichspargels der männlichen Hybridsorte `Gijnlim` sowie der gemischtgeschlechtlichen, anthocyanfreien Sorte `Huchels Schneewittchen` ergaben differenzierte Aussagen zur äußeren Qualität. Untersuchungen von Brix/ Zuckergehalt, Gehalt an Apfel-, Zitronen- und Weinsäure sowie proteinogenen Aminosäuren ( speziell essentielle und halbessentielle AS) und Gesamtpektin in den verschiedenen Spargelproben zeigten bei diesen wert- und geschmacksgebenden Inhaltsstoffen die Dynamik in ihrem Auftreten vom Erntezeitpunkt bis zum Auslagerungstermin auf. In diesem Zeitraum wurden auch der Frischgewichtsverlust erfasst und alle Untersuchungswerte stets vergleichend bei beiden Spargelanbauformen sowie beiden Sortentypen betrachtet. Dem bisherigen Kenntnisstand konnten einige neue Ergebnisse zur unterschiedlichen Qualitätsbildung und Qualitätserhaltungsmöglichkeit bei Grün- und Bleichspargel in Abhängigkeit von Sortenwahl, Bestandesdichte, Lagerverfahren und- dauer hinzugefügt werden. Sie bedürfen aber teilweise noch weiterer modifizierter Bearbeitung bzw. längerer Bearbeitungsdauer. / The quality is defined according to ISO 8402 as a whole of characteristics and significations of a product, which refer to their suitability for the fulfilment of fixed or presupposed requirements. Products with high water content and large breathing rate can suffer substantial quality impairments in the post-harvest period up to the time for selling and/or consumption. By the example of the asparagus, a rather young and physiologically very active plant rung, we register both external and internal breeding and plant-structural measures and storage procedures including precooling and duration of storage. The investigation material gathered in the Federal State of Brandenburg was won from a 2-factorial plot-field-test, a practice sort comparison test and by additional purchase. Bonitures and assortment after outwardly detectable, quality criteria both of freshly harvested and of two and/or five weeks under cool and APPROX. storage conditions (different gas compositions) of stored green and bleached Asparagus of the male hybrid sort ` Gijnlim` as well as the mixed-gender; Anthocyan-free sort ` Huchels Schneewittchen` resulted in differentiated statements to the outside quality. Investigations of Brix/ sugar content, content of apple -, lemon and tartaric acid as well as proteinogenen amino acids (particularly essential and halfessential AS) and total Pectin in the different asparagus samples pointed the dynamics in their occurrence from the harvest time to the paging date to taste-giving contents materials out with this worth and. In this period also the fresh decrease in weight and all investigation values always comparatively with both forms of cultivation of asparagus as well as both sort types were seized were regarded. The past level of knowledge could some new results for different quality formation and quality preservation possibility with green and bleaching asparagus as a function of sort choice, inventory density, camp procedure and duration be added. They require however partly still further modified treatment and/or longer machining time.

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.