Synthetic antioxidants have traditionally been added directly to food products in a single initial dose to protect against oxidation of lipids and generation of free radicals. Natural antioxidants have been shown to undergo loss of activity and become prooxidants at high concentrations; therefore, a need exists to develop active packaging which can gradually deliver antioxidants in a controlled manner. The objectives of this research were to (1) form and characterize cyclodextrin inclusion complexes with the natural antioxidants, alpha-tocopherol and quercetin, (2) incorporate cyclodextrin inclusion complexes of natural antioxidants into linear low density polyethylene (LLDPE), and (3) measure the release kinetics of inclusion complexes of natural antioxidants from LLDPE into a model food system. Cyclodextrin inclusion complexes of alpha-tocopherol and quercetin were formed by the coprecipitation method and characterized in the solid state by NMR, IR spectroscopy, and thermal analyses. Solid inclusion complex products of alpha-tocopherol:beta-cyclodextrin and quercetin:gamma-cyclodextrin had molar ratios of 1.7:1 as determined by UV spectrophotometry, which were equivalent to 18.1% (w/w) alpha-tocopherol and 13.0% (w/w) quercetin. Free and cyclodextrin complexed antioxidant additives were compounded with a twin-screw mixer into two LLDPE resin types followed by compression molding into films. Release of alpha-tocopherol and quercetin from LLDPE films into coconut oil at 30 °C was quantified by HPLC during 4 weeks of storage. The total release of alpha-tocopherol after 4 weeks was 70% from the free form and 8% from the complexed form averaged across both LLDPE resins. The mechanism by which alpha-tocopherol was released was modified due to its encapsulation inside the beta-cyclodextrin cavity within the LLDPE matrix as indicated by its diffusion coefficient decreasing by two orders of magnitude. Molecular encapsulation of natural antioxidants using cyclodextrins may be used as a controlled release mechanism within polymer food packaging to gradually deliver an effective antioxidant concentration to a food product, thereby, limiting oxidation, maintaining nutritional quality, and extending shelf life. / Ph. D.

Natamycin is an antimycotic compound that is widely used in the cheese industry to increase the shelf life of cheeses, especially shredded cheeses, by inhibiting the growth of molds. Natamycin is applied to the surface of cheese as an aqueous suspension or as a powder. However, natamycin is not readily water soluble making it harder to distribute evenly over shredded cheese Natamycin is degraded by ultraviolet (UV) light at wavelengths of 350 nm and below. Typical packaging applications do not provide adequate UV protection causing natamycin to degrade. This work was undertaken to determine the efficacy of UV absorber film to prevent UV light degradation of natamycin on the surface of shredded cheese. Current accepted methods to determine concentration of natamycin were evaluated for appropriateness in natamycin degradation studIes. The use of cyclodextrins to increase water solubility was tested to see if a uniform distribution of natamycin over the shredded cheese could be done effectively. Furthermore, a known application of mold was performed to see how well natamycin and each of its applications could prevent visible mold growth from occurring. The International Dairy Federation recognizes two methods to quantify natamycin on shredded cheese: high performance liquid chromatography (HPLC) and spectrophotometry. Concentrations of natamycin in aqueous suspensions were determined using both methods. Results show that spectrophotometry is flawed when quantifying the amount of active natamycin because the method gives erroneously high results. The amount of active natamycin is not accurately quantified using spectrophotometric techniques because it cannot separate the active form from the inactive form of natamycin. Polymer packages containing a UV absorber (11.4% light transmission at 350 nm) allow significantly less UV-associated degradation of natamycin than those packages that lacked a UV protectant (90.0% light transmission at 350 nm) (p<0.05). Incorporating a UV absorber into a package helps protect natamycin and its various complexes from UV light degradation, which can increase the shelf life of shredded cheese. However, even with a UV absorber, natamycin is still able to degrade. Natamycin was complexed with different cyclodextrins to help better solubilize natamycin â β-cyclodextrin, hydroxy-propyl β-cyclodextrin and γ-cyclodextrin. Using cyclodextrins to apply natamycin more uniformly onto shredded cheese did not significantly increase the consistency of distribution (p<0.05). Variability was uniform throughout all treatments with the exception of HPBCD complex. After 27 days, all of the UV packages treated with each of the cyclodextrin treatments containing shredded cheese began to show visible mold growth. Those packages stored in total darkness remained mold free through the duration of the experiment ending on day 62. When untreated with natamycin and an initial concentration of 101-102 spores/gram of Penicillium roqueforti, shredded cheese remained free from visible mold growth for 24 days in total darkness at 4°C. Samples treated with one of the natamycin treatments were able to remain mold free for at least 9 more days, showing visible signs of mold growth at day 33. There was no statistical difference between the treatments of dry natamycin, aqueous suspension natamycin, β-cyclodextrin-natamycin complex, and γ-cyclodextrin-natamycin complex (p<0.05). However, there was a difference with the use of hydroxy-propyl β-cyclodextrin-natamycin complex. Hydroxy-propyl β-cyclodextrin-natamycin complex allowed the shredded cheese to last for 41 days, 17 days longer than the control sample. / Master of Science

Odor detection thresholds of hexanal, 2-heptenal, 2-pentanone, and 2,4-nonadienal were determined in spring water, high temperature short time (HTST) 2% fat milk, and extended shelf life soymilk. The efficacy of odor scavenger's beta-cyclodextrin, D-sorbitol, and nylon 6 in removing these odors was also determined. The odor thresholds of the different odor and media combinations were as follows: hexanal in spring water, milk, and soymilk were 585, 339, and 536 ppb respectively; 2-heptenal in spring water, milk, and soymilk were 2,092, 2,322, and 3,184 ppb respectively; 2-pentanone in spring water, milk and soymilk were 24,925, 29,255 and 33,271 ppb respectively; and 2,4-nonadienal in spring water, milk, and soymilk were 164, 326, and 243 ppb respectively. These amounts reference the initial spiked concentration that was added directly to the media. Both hexanal and 2,4-nonadienal had lower thresholds than 2-heptenal and 2-pentanone in all of the media. The odor detection thresholds of 2-heptenal, 2-pentanone, and 2,4-nonadienal did show a significant difference between soymilk and water, but not for milk. The efficacy of the odor scavengers were determined by use of solid phase micro-extraction gas chromatography (SPME-GC) and sensory evaluation. Hexanal, 2-heptenal, 2-pentanone, and 2,4-nonadienal were spiked at 1,000, 3,000, 30,000, and 300 ppb respectively in all three media. Beta-cyclodextrin, D-sorbitol, and nylon 6 were added at a level of 0.1% w/v and 1.0% w/v. In all of the media, beta-cyclodextrin was found to significantly reduce hexanal, 2-pentanone, 2-heptenal, and 2,4-nonadienal at both 0.1% w/v and 1.0% w/v. Nylon 6 was not found beneficial. / Master of Science

Natamycin is an antimycotic with very low water solubility and extremely high photosensitivity, which is used to extend the shelf life of shredded cheese products. The objectives of this research are: (a) to find a new delivery system for natamycin, which increases its aqueous solubility and (b) to increase the chemical stability of natamycin so that it has a prolonged antifungal effect on the surface of the shredded cheese. Molecular inclusion complexes of natamycin were formed with β-, hydroxypropyl β-, and γ- cyclodextrins (CDs) which allowed large increases in aqueous solubility without the use of organic co-solvents or surfactants. The water solubility of natamycin was increased 16-fold, 73- fold, and 152-fold with β-CD, γ-CD, and hydroxypropyl β-CD, respectively. The natamycin:CD inclusion complexes resulted in nearly equivalent in vitro antifungal activity as natamycin in its free state. Nuclear magnetic resonance (NMR) was utilized to prove the formation of true inclusion complexes. 1H NMR shift titrations of N-(3 -N-dimethylaminosuccimido) natamycin with β- and γ-CDs enabled determination of the stoichiometry of both complexes as 1:1. Aqueous solutions of natamycin (20 mg/L) were found by quantitative HPLC to be completely degraded after 24 hours of exposure to 1000 lux fluorescent lighting at 4 °C. After 14 days of storage in darkness at 4 °C, 92.2% of natamycin remained in active form. Aqueous solutions of natamycin:β-CD complex and natamycin:γ-CD complex were significantly more stable (p < 0.05) than natamycin in its free state when stored in darkness at 4 °C. Clear poly(ethylene terephthalate) packaging with an ultraviolet light absorber allowed 85.0% natamycin to remain after 14 days of storage under 1000 lux fluorescent lighting at 4 °C. Such dramatic increases in water solubility and light stability will enable natamycin to function as a more effective antimycotic in the food industry. / Master of Science

Tesis por compendio / Starch is the major energy reserve in plants, extensively present in many food and non-food applications, and is one of the most abundant carbohydrates in human diet. In addition to starch native form currently used as a raw material for industrial applications, modified starches have become very attractive to develop numerous products that have greatly expanded starch use and utility. Enzymatic modifications are carried out to enhance starch functionality with the aim of overcoming technological constraints and to improve final product quality. Above all starches, corn starch is widely produced and consumed, and used as main ingredient to produce gluten-free products. This Thesis focuses to the study of the individual effect of three different enzymes, fungal alpha-amylase, amyloglucosidase and cyclodextrin glycosyltransferase, on the above sub-gelatinization temperature of corn starch, in order to modify its properties and extend its applications. Different analyses have been performed to improve the understanding of functional and structural changes promoted by the enzyme action. Moreover, corn starch modified with cyclodextrin glycosyltransferase was selected to investigate the glycemic response in mice. The presence/absence of hydrolysis products released from the enzyme catalytic activity and gelatinization properties were related to glycemic index. Enzymatically modified corn starch resulted in a diversity of functional starch granules that may be used in food applications for many purposes. / El almidón es la principal fuente de reserva de energía en las plantas, está ampliamente presente en diversas aplicaciones alimentarias y no alimentarias, y constituye uno de los hidratos de carbono más abundantes en la dieta humana. Además del uso frecuente y clásico del almidón nativo como materia prima en la producción de alimentos, los almidones modificados han experimentado gran expansión en el desarrollo de numerosos productos debido a su carácter versátil. Las modificaciones enzimáticas se llevan a cabo para realzar la funcionalidad del almidón con objeto de soslayar las restricciones tecnológicas y mejorar la calidad del producto final. En concreto, el almidón de maíz ampliamente producido y consumido, es apropiado como ingrediente principal en la producción de alimentos libres de gluten. Esta Tesis aborda el estudio del efecto individual de tres diferentes enzimas, alfa-amylasa fúngica, amiloglucosidasa y ciclodextrina glucosiltransferasa, sobre la temperatura de sub-gelatinización del almidón de maíz. Se realizaron diferentes análisis para ampliar el conocimiento de los cambios estructurales y funcionales asociados a la acción de la enzima. Así mismo, se seleccionó el almidón de maíz modificado con ciclodextrina glucosiltransferasa para investigar la respuesta glicémica en ratones. El índice glicémico se relacionó con la ausencia/presencia de productos de hidrólisis liberados por la acción catalítica de la enzima y las propiedades de gelatinización. Los almidones de maíz enzimáticamente modificados presentaron alteraciones funcionales en los gránulos de almidón, lo que les confiere características de interés para diversos usos alimentarios. / El midó és la principal font de reserva d'energia en les plantes, està àmpliament present en diverses aplicacions alimentàries i no alimentàries, i constitueix un dels hidrats de carboni més abundants en la dieta humana. A més de l'ús freqüent i clàssic del midó natiu com a matèria primera en la producció d'aliments, els midons modificats han experimentat gran expansió en el desenvolupament de nombrosos productes a causa del seu caràcter versàtil. Les modificacions enzimàtiques es duen a terme per a realçar la funcionalitat del midó a fi d'esbiaixar les restriccions tecnològiques i millorar la qualitat del producte final. En concret, el midó de dacsa àmpliament produït i consumit, és apropiat com a ingredient principal en la producció d'aliments lliures de gluten. Aquesta Tesi aborda l'estudi de l'efecte individual de tres diferents enzims, alfa-amylasa fúngica, amiloglucosidasa i ciclodextrina glucosiltransferasa, sobre la temperatura de sub-gelatinizació del midó de dacsa. Es van realitzar diferents anàlisi per a ampliar el coneixement dels canvis estructurals i funcionals associats a l'acció de l'enzim. Així mateix, es va seleccionar el midó de dacsa modificat amb ciclodextrina glucosiltransferasa per a investigar la resposta glicémica en ratolins. L'índex glicémic es va relacionar amb l'absència/presencia de productes d'hidròlisis alliberats per l'acció catalítica de l'enzim i les propietats de gelatinización. Els midons de dacsa enzimàticament modificats van presentar alteracions funcionals en els grànuls de midó, la qual cosa els confereix característiques d'interès per a diversos usos alimentaris. / Durá De Miguel, Á. (2017). EFFECT OF ENZYMATIC TREATMENTS ON CARBOHYDRATE MATRICES TOWARDS HEALTHY GLUTEN FREE FOODS APPLICATION [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/79509 / Compendio

Cinnamic acid (CA) is a naturally occurring organic acid that is found in some fruits and a number of spices. CA has antimicrobial activity against certain spoilage microorganisms and pathogenic bacteria. However, the acid is poorly soluble in water. Cyclodextrin molecules have a hydrophobic cavity that allows them to serve as a host for insoluble molecules in aqueous matrices. This study was conducted to determine if the aqueous solubility of cinnamic acid could be improved via complexation with α- or β-cyclodextrins, and if these complexes could be used to control bacterial pathogens in juices. Based upon phase solubility analysis, α-cyclodextrin was chosen as the host molecule for the remainder of this study. In complex with α-cyclodextrin, the solubility of cinnamic acid increased from approximately 400 mg/L to 3800 mg/L. Prepared cinnamic acid complexed with α-cyclodextrin was aseptically added (400 mg/L and 1000 mg/L) to orange juice inoculated with a Salmonella enterica (7 log CFU/mL) and apple cider inoculated with Escherichia coli O157:H7 (7 log CFU/mL). Cider and orange juice samples were extracted on day 0 and at 24 h intervals for seven days and spread plated onto Tryptic Soy Agar. Cinnamic acid was effective for reducing populations of both bacterial pathogens in juice. Populations of E. coli O157:H7 in the apple cider were significantly reduced after 7 days at 25.6 ± 0.42°C at concentrations of 400 mg/L (5-log CFU/mL reduction) and 1000 mg/L (6-log CFU/mL reduction) cyclodextrin-cinnamic acid. S. enterica counts were also reduced in orange juice at 4° C treated with 400 mg/L (2.7-log CFU/mL reduction) and 1000 mg/L (3.2-log CFU/mL reduction) complexed cinnamic acid. The much improved solubility of this compound provides food processors with greater flexibility in using cinnamic acid in their product formulations. / Master of Science

This thesis explores the structures and structural changes of supramolecular host-guest systems, proteins, and other small molecules in the gas phase, utilizing a combination of computational modeling and experimental data. The primary instruments employed were a Fourier transform ion cyclotron resonance mass spectrometer (FTICR-MS) and an ion mobility mass spectrometer (IM-MS). In the IM-MS experiments, the focus was on investigating the binding behavior of cyclodextrin macrocycles—specifically α, β, and γ-cyclodextrin—with per-fluoroalkane substances (PFAS), which are pervasive environmental contaminants. This investigation involved measuring ion-neutral collision cross sections and using computational modeling to determine whether PFAS compounds bind inside or outside the cyclodextrin cavity. The results indicate that only β-cyclodextrin binds PFAS compounds internally, attributed to its seven-fold symmetry and the localized hydrogen bonding network across the macrocycle's secondary face. Conversely, α and γ-cyclodextrin appear to favor collapsing inward, enhancing internal hydrogen bonding while keeping the PFAS bound externally. The FTICR-MS instrument was used for time-resolved CRAFTI (TR-CRAFTI) collision cross section measurements on various systems, including tetraalkylammoniums (TAA), cytochrome C, and β-cyclodextrin host-guest complexes. This involved activating gas-phase ions using sustained off-resonance irradiation (SORI) activation, followed by a variable delay for collisional cooling. Subsequently, a CRAFTI measurement was conducted to obtain a timeresolved view of the collision cross section. Initial findings suggest the feasibility of measuring and modeling structural changes post-activation over varying time scales, ranging from approximately 100 milliseconds to 10 seconds, depending on the size and complexity of the system being studied.

Wood-based panel products (WBPs) are considered highly sustainable due to their carbon neutrality, low embodied energy, and ability to be produced from various low-grade wood species. However, WBPs exhibit limited resistance to biological degradation, which can deteriorate their mechanical performance and shorten their service life. Additionally, WBPs may not be fully compatible with traditional wood preservation chemicals. This dissertation demonstrated a facile method of synthesizing essential oil-based natural preservatives compatible with WBPs. Four essential oils (EOs), namely thymol (THY), trans-cinnamaldehyde (tCN), carvacrol (CRV), and eugenol (EUG), were complexed with beta-cyclodextrin (beta-CD) to form beta-CD-EOs inclusion complexes through a sonochemical process using a high-power ultrasonicator (750W). The sonochemical variables, such as a molar ratio between beta-CD and EO, ultrasonication power, and centrifugation speed and time, were optimized to improve the inclusion yield of EOs in the complex and facilitate the large-scale production of beta-CD-EOs. Attenuated Fourier transform infrared spectroscopy (AT-FTIR) and X-ray diffraction analysis confirmed the formation of inclusion complexes at the different production scales. The inclusion yield of EOs in the complexes was estimated by ultraviolet-visible spectrophotometry (UV-vis) as well as thermogravimetric analysis (TGA), which was capable of differentiating unreacted EOs and loosely encapsulated EO molecules from the estimation of the inclusion yield. Differential scanning calorimetry (DSC) revealed that beta-CD-tCN did not adversely affect the curing properties of polymeric methylene diisocyanate (pMDI) used as a glue for plywood. The influence of beta-CD-tCN addition on the bonding strength of the wood composite was assessed by a lap shear strength test using 3-ply SYP plywood, following ASTM D906-20. Statistical analysis using the Tukey test at p<=0.05 indicated no significant difference between the bonding strength of pMDI and beta-CD-tCN as an additive, indicating that beta-CD-tCN could be successfully implemented in the manufacturing process of wood composite products as natural preservatives.

>Magister Scientiae - MSc / Glucose oxidase (GOx) and horseradish peroxidase (HRP) are important enzymes for the development of amperometric enzyme linked immunosensors. The selectivity of each enzyme towards its analyte deepens its importance in determining the sensitivity of the resultant immunosensor. In designing immunosensors that have customized transducer surfaces, the incorporation with FAD and iron based enzymes ensures that electron kinetics remains optimal for electrochemical measurement. Various different immobilization strategies are used to produce response signals directly proportional to the concentration of analyte with minimal interferences. The combination of self-assembled monolayers and supramolecular chemistry affords stability and simplicity in immunosensor design. In this work, two electrochemical strategies for the detection of human chorionic gonadotropin(hCG) is presented. This involves the modification of a gold surface with a thiolated β-cyclodextrin epichlorohydrin polymer (βCDPSH) to form a supramolecular inclusion complex with ferrocene (Fc)-functionalised carboxymethyl cellulose polymer (CMC). Cyclic voltammetry indicated that ferrocene is in close proximity to the electrode surface due to the supramolecular complex formed with βCDPSH. Furthermore, strategy (a) for the detection of hCG used α-antihCG labelled (HRP) as reporter conjugate. Strategy (b) maintained the CMC bifunctionalised with Fc and recognition antibody for hCG hormone. However, the system was functionalised with a HRP enzyme and detection is done by using GOx reporter conjugates for in situ production of hydrogen peroxide. The reduction of H2O2 was used for the amperometric detection of hCG by applying a potential of 200 mV. The sensitivity and limit of detection of both strategies were calculated from calibration plots. For strategy (a) the LOD was found to be 3.7283 ng/mL corresponding to 33.56 mIU/mL and a sensitivity of 0.0914 nA ng-1 mL-1. The corresponding values for strategy (b) are 700 pg/mL (6.3 mIU/mL) and 0.94 nA ng-1 mL-1.

Die Kombination aus anorganischen Nanopartikeln und Mikrogelen in einem hybriden System erlaubt die Herstellung von Materialien mit vielseitigen neuen Eigenschaften. Im Idealfall weisen solche hybriden Materialien neben den Eigenschaften von beiden indivduellen Systemen zusätzlich synergetische Effekte auf, welche aus den Interaktionen zwischen dem anorganischen Nanopartikel und dem Mikrogel resultieren. Im ersten Teil dieser Arbeit wird eine neuartige und eingängige Methode zur Herstellung von Cu2O@PNIPAM Kern-Schale Nanoreaktoren präsentiert. Die PNIPAM Schale schützt dabei die Cu2O Nanopartikel effektiv vor Oxidation. Die Cu2O@PNIPAM wurden als Photokatalysator zum Abbau von Methylorange unter sichtbarem Licht eingesetzt. Im Vergleich zu den reinen Cu2O Nanopartikeln konnte eine signifikante Steigerung der katalytischen Aktivität festgestellt werden. Desweiteren kann die photokatalytische Aktivität mittels Temperatur durch die thermosensitive PNIPAM Schale abgestimmt werden. Verhältnismäßig geringe Konzentrationen einer Cu2O@PNIPAM wässrigen Lösung (1,5 Gew%) können direkt als neuartige Tinte genutzt werden. Keine zusätzlichen Additive oder organische Lösungsmittel sind für die Strahldruckprozesse vonnöten. Gedruckte Bauelemente bestehend aus den Cu2O@PNIPAM wurden als Gas Sensoren eingesetzt und zeigten eine geringere Nachweisgrenze für NO2 als die reinen Cu2O Nanowürfel. Im zweiten Teil der Arbeit wurden katalytisch aktive Au Nanopartikel an copolymerisierten α –Cyclodextrin (α-CD) Einheiten in einem Poly(N-vinylcaprolactan) (PVCL) Mikrogel immobilisiert. Diese hybriden Partikel sind sehr aktive Katalysatoren für die Reduktion von aromatischen Nitroverbindungen. Die Reduktion von 4-Nitrophenol (Nip) und 2,6-Dimethyl-4-nitrophenol (DMNip) wurden als Modellreaktionen ausgewählt. Durch selektive Bindungseingenschaften der Nitroverbindungen an die α-CD Einheiten konnten verschiedene katalytische Aktivitäten für Nip and DMNip festgestellt werden. / The combination of inorganic nanoparticles and organic microgels in one hybrid system allows for the preparation of new materials with multifunctional properties. Ideally, such hybrid materials reflect both the properties of its individual components and synergetic effects due to the interaction between inorganic nanoparticles and microgels. In the first part of this thesis, the fabrication of Cu2O@Poly(N-isopropylacrylamide) (PNIPAM) core-shell nanoreactors has been presented. It was found that the PNIPAM shell effectively protects the Cu2O nanocubes from oxidation. The core-shell microgels have been used as photocatalyst for the decomposition of methyl orange and a significant enhancement in the catalytic activity has been observed compared with the bare Cu2O nanocubes. Most importantly, the photocatalytic activity of the core-shell nanoreactors can be further tuned by the thermosensitive PNIPAM shell. The aqueous solution of Cu2O@PNIPAM core-shell nanoparticles with quite low solid content (1.5wt. %) can be also directly used as a novel ink material for the inkjet printing without adding any other surfactants and organic solvents. The gas sensor device printed by core-shell nanoparticles is more sensitive to NO2 than that made from the bare Cu2O nanocubes. In the second part, a kind of hybrid microgel has been fabricated by immobilization of catalytically active Au nanoparticles in the α-cyclodextrin (α-CD) modified poly(N-vinylcaprolactam) (PVCL) microgels without addition of reducing agent and surfactant. The hybrid microgels can work efficiently as catalyst for the reduction of aromatic nitro-compounds by using the reduction of 4-nitrophenol (Nip) and 2,6-dimethyl-4-nitrophenol (DMNip) as model reactions. Due to the selective binding property of α-CDs to nitro compounds, the synthesized hybrid microgels show different catalytic activity for the target compounds, 4-nitrophenol (Nip) and 2,6-dimethyl-4-nitrophenol (DMNip), during the catalytic reactions.