With the growing concern of the accumulation of plastic-based food packaging waste, the search for bio-based biodegradable packages is on the rise. These materials differ from their petro-based counterparts in their degradation rates, which are much higher in the former. Not only do bio-based biodegradable materials degrade faster during post-usage processes but also they age faster during usage and storage, which affects their performance and functionality. The application of noninvasive testing methods with the capability to report on the matrix’s state could assist in the development of a more ubiquitous way to assess ageing in food packaging, particularly in biodegradable ones. To this end, the performance of a luminescence spectroscopy technique based on three luminescent probes, one intrinsic to the matrix and two added, was monitored and the sensitivity of the probes to report on ageing was analyzed. Biodegradable films were made of 2% gelatin (type A) and 0.5% glycerol (plasticizer). Gelatin contains an intrinsic fluorophore, the aromatic amino acid tyrosine (Tyr), which can report on the molecular mobility of a matrix. Additionally, the films were doped with two extrinsic fluorophores, Fast Green FCF (FG) at 0.124 mM and pyranine (Pyr) at 0.05 mM, which can report on the physical state and available free water within a matrix, respectively. Films were casted onto plastic Petri dishes and stored at five relative humidities (RHs), namely 2.5, 25, 33, 53, and 75%, for five weeks with measurement collection every week. Films were tested using fluorescence spectroscopy at excitation and emission range wavelengths optimized depending on the assessed probe. Additional measurements to determine moisture content, changes in secondary protein structure using FTIR spectroscopy, and mechanical properties using a Universal Testing Machine in tensile mode aided in the evaluation of the sensitivity of the luminescent probes in sensing ageing. Luminescent probes, intrinsic or added, have the capability to assess the physical state of the films in situ and can provide molecular level sensing of their local environment. Tyr emission showed a sharp increase in fluorescence intensity in films stored at low RH as a function of time. FG showed a similar pattern to that of Tyr but higher sensitivity to changes along the observed period. The two characteristics emission bands of Pyr provide information on the state of water within the matrix. Although the results on this probe hinted microstructural rearrangements within the films as a function of time, the sensitivity of this probe was not high enough at the conditions evaluated and provided limited information on films’ solvation. The sensitivity of the luminescent probes to changes during ageing were revealed through correlation of the photophysical properties of the two effective probes, Tyr and FG, and the mechanical properties of the films at different RH through storage. Both methods, mechanical and optical, were similarly sensitive to changes during ageing particularly after 3-week storage. However,, it can be speculated that because of the different scales at which optical and mechanical measurements report (local vs. bulk), the methods, they could also complement each other. These findings suggest that, in principle, a luminescence spectroscopy technique using intrinsic and extrinsic probes can replace mechanical testing to noninvasively monitor structural changes and stability of biodegradable packaging as a function of time.

<p>Naturally occurring carbohydrates polymers and their functional derivatives play important roles in the research and technology development in the food, nutrition, and pharmaceutical areas. A major property of these polymeric materials is to associate, enable, enhance, and/or deliver small-molecule active compound such as phytochemicals, nutraceuticals, and active pharmaceutical ingredients (APIs). The goal of this project was to synthesize and characterize phytoglycogen-based materials and study their structure-function relationships in association with selected small-molecule active compounds, including resveratrol, a food-related poorly water-soluble phenolic compound, griseofulvin, an insoluble API, and CCVJ (9-(2-carboxy-2-cyanovinyl) julolidine) a molecular rotor used as a structural probe of polymeric materials. </p><p>In this study, phytoglycogen (PG) was derivatives to phytoglycogen octenyl succinate (PG-OS), hydroxypropyl phytoglycogen (HPP), and octenylsuccinate hydroxypropyl phytoglycogen (OHPP). PG, HPP, and OHPP were evaluated for their efficacy in improving the solubility and Caco-2 permeation of resveratrol and griseofulvin, and using CCVJ, PG-OS was evaluated on its performance at oil-water interface in comparison with OSA-starch, acacia gum, and sodium caseinate. The results showed that: 1) PG, HPP, and OHPP substantially improved the soluble amount and Caco-2 monolayer permeation of resveratrol and griseofulvin, and anti-fungal efficacy of griseofulvin in the aqueous system were significantly enhanced; suggesting that the active ingredients were effective solubilized and released to become bioavailable, 2) among all PG-based biopolymers, OHPP showed superior performance in solubilizing resveratrol and griseofulvin, and 3) in the oil-water two-layer model system, PG-OS, OSA-starch, acacia gum, and sodium caseinate all affected the transferring of CCVJ from oil to aqueous phase, and the effect was monitored and interpreted by the emission spectra of molecular rotor; in the emulsion system, the emission peak wavelength of CCVJ was correlated with the amount of biopolymer adsorbed at the interface of emulsion droplets, and the molecular rotor-based method can be used to characterize the interfacial adsorption of biopolymer at the interface in oil-in-water emulsion.</p><p>This study provides information on the interactions between phytoglycogen-based biopolymers and poorly water-soluble active ingredients, and may potentially supports the study of new functional ingredients interaction with phytoglycogen-based biopolymers in aqueous system. Furthermore, this work allowed us to advance the use of molecular rotor as new analytical tool to study the physicochemical properties of biopolymer.</p>

<p>The discovery and utilization of novel starches with unique superb properties are highly demanded for modern industrial uses. Small-granule starch (SGS) is a category of unconventional starches with the granular size smaller than 10 μm. The potential use of SGS includes many conventional and novel high-value applications, such as texturizing, fat replacement, encapsulation, controlled delivery and nano-engineering. In the present work, we focused on three SGS isolated from amaranth (<i>Amaranth cruentus</i>), cow cockle (<i>Saponaria vaccaria</i>) and sweet corn (<i>sugary-1</i> maize mutant). The basic structural and unique physical properties of SGS were characterized and compared to common large-granule food starches. It was found that (1) the highly branched amylopectin contributed to low crystallinity and pasting viscosities of sweet corn starch, (2) cow cockle starch exhibited high shear-resistance and low retrogradation in prolonged storage, and (3) the amylopectin for amaranth starch was less branched with small clusters, which was associated with the high crystallinity, medium shear-resistance and low pasting viscosity of amaranth starch. Despite the small size of starch granules, SGS in both native and swelling states showed the capacity of retaining small molecules. Compared to large-granule starch, native SGS are more difficult for small molecules to reach an equilibrium permeation. This work provides insights to the fine structure and physicochemical behaviors of selected high-potent SGS, which is believed to support the industrial production and application of SGS in the future.</p> <p>The characteristics of local polymeric structure dominate many critical properties of glucan particles, such as starch retrogradation and the loading and stabilizing of active substance. Molecular rotor (MR), a fluorescent probe, was proposed to fulfill the simple, high-sensitive, and quantitative-based characterization of local glucan architecture (LGA). In the present work, two innovative studies relevant to this novel method were conducted: (1) MR was able to characterize glucans based on its unique fluorescent response to characteristic LGA, (2) MR was able to sensitively probe and visually demonstrate the transition of LGA induced by starch retrogradation. This novel MR-based approach is expected to advance carbohydrate-related researches in the future.</p>

The primary focus of this dissertation is the development of novel fluorescent near-infrared molecules for various applications. In Chapter 1, a compound dU-BZ synthesized via Sonogashira coupling reaction methodology is described. A deoxyuridine building block was introduced to enhance hydrophilic properties and reduce toxicity, while an alkynylated benzothiazolium dye was incorporated for near-IR emission and reduce photodamage and phototoxicity that is characteristic of common fluorphores that are excited by UV or visible light. A 30-fold enhancement of fluorescence intensity of dU-BZ was achieved in a viscous environment. Values of fluorescence quantum yields in 99% glycerol/1% methanol (v/v) of varying temperature from 293 K to 343 K, together with fluorescence quantum yields, radiative and nonradiative rate constants and fluorescence lifetimes in glycerol/methanol solutions of varying viscosities from 4.8 to 950 cP were determined. It was found that both fluorescence quantum yields and fluorescence lifetimes increased with increasing viscosity, which is consistent with results predicted by theory. This suggests that the newly designed compound dU-BZ is capable of functioning as a probe of local microviscosity, and was later confirmed by in vitro bioimaging experiments. In Chapter 2, a new BAPTA (O,O*-bis(2-aminophenyl)ethyleneglycol-N,N,N*,N*-tetra acetic acid) and BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene)-based calcium indicator, BAPBO-3, is reported. A new synthetic route was employed to simplify both synthesis and purification, which tend to be low yielding and cumbersome for BAPTA derivatives. Upon excitation, a 1.5-fold increase in fluorescence intensity in buffer containing 39 ?? Ca2+ and a 3-fold increase in fluorescence intensity in buffer containing 1 M Ca2+ was observed; modest but promising fluorescence turn-on enhancements. In Chapter 3, a newly-designed unsymmetrical squaraine dye, SQ3, was synthesized. A one-pot synthesis was employed resulting in a 10% yield, a result that is generally quite favorable for the creation of unsymmetrical squaraines Photophysical and photochemical characterization was conducted in various solvents, and a 678 nm absorption maximum and a 692 nm emission maximum were recorded in DMSO solution with a fluorescence quantum yield of 0.32. In vitro cell studies demonstrated that SQ3 can be used as a near-IR probe for bioimaging.

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No description available.

<p>Maize is one of the top sources of food starch. Industrial use of starch is mostly in its native form and used due to their functional and structural properties. Native starch properties and functionalities have been altered using chemical. An alternative for the development of native starch substituents with desirable starch properties is the use of mutagenesis techniques to increase genetic variation in maize kernels. With this approach, a highly diverse library of native starches with different properties are produced. Traditional analysis of the functional and structural properties requires generous amounts of material as well as a time-consuming and costly breeding process to obtain enough kernels. To address this difficulty, high-throughput techniques are proposed for studying starch properties and functions which includes a 1) single kernel sampling method for the isolation of milligrams of starch, and techniques for studying starch based on functional properties, 2) retrogradation and 3) shear resistance, using low-volume low-concentration starch pastes.</p><p>First, three mechanical approaches were evaluated for the collection of endosperm samples from individual kernels: razor blade, 1.5 mm drill bit, and trephine bur. Furthermore, two methods for the isolation of crude starch from endosperm samples (steeping method and combination of proteases and sonication) were compared. In this study, the mechanical approaches were evaluated using the recovery rate, throughput, and germination rate of sampled kernels. Moreover, yield determination, particle size distribution, and morphological evaluation using a light microscope were performed on crude starch isolated from the endosperm samples. The use of trephine bur to collect endosperm samples and isolation of crude starch using protease digestion and sonication showed the best combination for a high-throughput setting. </p><p>Second, a high-throughput technique using milligram sample for the screening of retrogradation-resistant starch was evaluated by comparing two spectrophotometric techniques: turbidity method and molecular rotor (MR). MRs are fluorescent probes with high sensitivity to the viscosity of their environment, polarity of the media, molecular crowding, and free volume. After excitation, MRs relax through rotational movement and reduces the emission of fluorescence. In this study, hydroxypropylated waxy corn starch (WCS) and hydroxypropylated normal corn starch (NCS) were used and their retrogradation kinetics was compared with retrogradation kinetics of native WCS and NCS. </p><p>It was found that the molecular rotor 9-(2-carboxy-2-cyaovinyl)-julolidine (CCVJ) was effective to sense changes during slow retrogradation of amylose-containing starch pastes. Development of elastic modulus of retrograded NCS pastes obtained from dynamical rheology showed high correlation with the development of fluorescence intensity of the CCVJ. Furthermore, rate of retrogradation using fluorescence intensity was affected by the introduction of a retrogradation inhibitor, hydroxypropyl groups. Accelerated retrogradation of low-concentration WCS pastes was measured using the turbidity method and fluorescence intensity of CCVJ in a microplate. Accelerated retrogradation was performed by subjecting the low-concentration WCS pastes to six freeze-thaw cycles of -20 ºC for 1 hour and 30 ºC for 1 hour. Overall, development of turbidity resulted in the more sensitive technique to detect rate of retrogradation of amylopectin-containing starch. </p><p>The last part of this research studied the use of CCVJ as a technique to identify shear-resistant starch in starch slurries using milligram sample. For this purpose, WCS was cross-linked with sodium trimetaphosphate (STMP) and phosphoryl chloride (POCl₃). Low-volume starch slurries having CCVJ were prepared ranging from 0.5% to 1% starch concentration in a 96-well PCR plates and subjected to heat and shear treatments. It was found that fluorescence intensity measured in native WCS pastes were the lowest. Furthermore, fluorescence intensity of the CCVJ in the gelatinized starch increased as the amount of cross-linker increased in the cross-linked WCS. After shear treatments, the same trend in fluorescence intensity increase was recorded in all the crosslinked WCS. Results obtained using fluorescence intensity were compared with rapid viscosity analyzer (RVA) and images from microscope. Results obtained from both techniques corroborated the findings using fluorescence intensity.</p><p>In general, the findings of this research provide new insights into the possibilities of developing a high-throughput screening platform of milligram starch sample based on their physical properties. </p>