The presented thesis on thermosensitive polymer gel is focused especially on a thermosensitive triblock copolymer, which is composed of hydrophobic polylactide, polyglycolid and hydrophilic polyethylene glycol (PLGA-PEG-PLGA). Thermosensitive copolymers are very attractive for their phase sol-gel transitions and gel-suspension transitions. The aqueous solution of this copolymer behaves like a sol at laboratory temperature and like a gel at body temperature. These systems are used as injectable carriers for targeted drug delivery with controlled release. However, the influence of the resulting polymer concentration and temperature on the thermosensitive hydrogel nanostructure was not yet fully studied. In the experimental part, the viscoelastic behavior of hydrogels was observed by dynamic rheological analysis at different polymer concentrations and temperature conditions. The average size and distribution of micelles of triblock copolymer in aqueous solution were measured using dynamic light scattering technique. Characterization of fibrous micelles was complemented by imaging technique, cryogenic transmission electron microscopy.

This diploma thesis deals with the determination of glucosinolates in plant material by capillary electrophoresis and high performance liquid chromatography. The chemical structure, biosynthesis, degradation, and also biological effects of glucosinolates are described. One part of this work also deals with the methods, which glucosinolates in plant materials were determined by. The experimental part describes the separation of intact glucosinolates by capillary electrophoresis and high performance liquid chromatography. Two plant materials were available for the determination of glucosinolates, namely lyophilized rapeseed leaves and broccoli juice. Micellar electrokinetic chromatography using a cationic surfactant was used to determine intact glucosinolates by capillary electrophoresis. After finding the optimal conditions for the separation of intact glucosinolates, it was found that it is impossible to determine these substances in plant samples. The reason was interference from the matrix, which interfered with this determination. While using high performance liquid chromatography under optimal conditions, some of the intact glucosinolates were identified in a rapeseed plant sample. Furthermore, the calibration dependencies of individual glucosinolates were obtained and the recovery and...

The Western Dairy Center at Utah State University funded this project to investigate cheese research using concentrated milks. Concentrated milk was provided by the South Dakota State University and starter culture for this study was prepared and donated by Vivolac Cultures Corporation, Greenfield, Indiana. The project initiated as a continuation of a previous study on effects of protein concentration, coagulum cut size and set temperature on curd moisture loss kinetics while stirring during cheesemaking. It was aimed at determining the extent to which pH drop prior to draining and final cheese moisture when using microfiltered concentrated milk. We performed twelve cheesemaking trials using recombined milk from micellar casein concentrate, cream and skim milk according to a modified cheddar cheese-make procedure. Four different levels of starter cultures were used to achieve different acidification rates for pH change during cheesemaking. The amount of starter culture added had significant effect on moisture of cheese at whey drainage, moisture and pH of cheese. Thus, it can be said that the pH drop that occurs during the cheesemaking increases rate and extent of whey expulsion.

Micellar electrokinetic capillary chromatography (MECC) is a form of capillary zone electrophoresis. Addition of a surfactant produces micelles in an aqueous/organic buffer. Separation of drugs is obtained via differences in the electrophoretic mobilities of the analytes within the capillary, resulting from their electrophoretic velocity and the electroosmotic flow of the buffer in a given electric field. The migration order is determined by the differential partitioning of the drugs between the micelles and the aqueous/organic phase. Barbiturates were extracted from various biological fluids at pH 4.5 with TOXI-TUBES B. MECC analyses were performed using a Waters Quanta 4000 Capillary Electrophoretic System with a 745 Data Module with a 75 μ x 60 cm capillary and an aqueous/organic buffer of 85% 10 mM borate, 10 mM phosphate, 100 mM sodium dodecyl sulfate and 15% acetonitrile at a pH of 8.5 with a voltage of 20 kV using ultraviolet absorption detection at 214 nm. Migration times were: phenobarbital, 7.78 min.; butalbital, 8.01 min.; butabarbital, 8.23 min.; mephobarbital (internal standard), 8.88 min.; amobarbital, 9.41 min.; pentobarbital, 10.03 min. and secobarbital, 10.79 min. Correlation coefficients (r) between peak areas and concentration ranges of 3 to 60 μg/mL were from 0.964 to 0.999. Coefficients of variation (CV) raged from 2.6 to 8.6% between days and 2.3 to 9.8% within day. Application of this methodology to four forensic cases of butalbital intoxication detected concentrations of 0.7 to 12.7 μg/mL in blood; 0.8 to 1.9 μg/mL in vitreous humor and 1.5 to 7.6 μg/mL in urine. MECC is applicable to forensic analysis of barbiturates extracted from biological fluids.

No description available.

Capillary electrophoresis (CE) is an analytical chemistry approach that allows for the efficient separation by charge of diverse classes of compounds for analysis, including secondary metabolites. The goal of this work was to optimize a buffer system for plant tissue analysis using micellar electrokinetic chromatography (MEKC), and by doing so to understand the role of buffer components in the performance of this form of capillary electrophoresis. In this experiment we implemented a factorial design to optimize buffer composition for separating plant tissue and secondary metabolites. The results of this experiment will be used to optimize a universal buffer for MEKC analysis that can be used on any variety of plant tissues. To determine the feasibility of this, a diverse set of plant secondary metabolite chemical standards in solution were tested as well as Helianthus annuus tissue to confirm the separation in a real biological sample. The results of this optimization yield insights into the utility of buffer components like electrolyte and pH for MEKC separation.

No description available.

Micellar electrokinetic chromatography (MEKC), one of the major capillary electrophoresis (CE) modes, has been interfaced to mass spectrometry (MS) to provide high sensitivity and selectivity for analysis of chiral compounds. The research in this dissertation presents the development of novel polymeric glucopyranoside based molecular micelles (MoMs) (aka. polymeric surfactants) and their application in chiral MEKC-MS. Chapter 1 is a review of chiral CE-MS - in the period 2010-2015. In this chapter, the fundamental of chiral CE and CE-MS is illustrated and the recent developments of chiral selectors and their applications in chiral EKC-MS, CEC-MS and MEKC-MS are discussed in details. Chapter 2 introduces the development of a novel polymeric α-D-glucopyranoside based surfactants, n-alkyl-α-D-glucopyranoside 4,6-hydrogen phosphate, sodium salt. In this chapter, polymeric α-D-glucopyranoside-based surfactants with different chain length and head groups have been successfully synthesized, characterized and applied as compatible chiral selector in MEKC-ESI-MS/MS. or the enantioseparation of ephedrines and β-blockers. Chapter 3 continues to describe the employment of polymeric glucopyranoside based surfactants as chiral selector in MEKC-MS/MS. The polymeric β-D-glucopyranoside based surfactants, containing charged head groups such as n-alkyl β-D-glucopyranoside 4,6-hydrogen phosphate, sodium salt and n-alkyl β-D-glucopyranoside 6-hydrogen sulfate, monosodium salt were able to enantioseparate 21 cationic drugs and 8 binaphthyl atropisomers (BAIs) in MEKC-MS/MS, which promises to open up the possibility of turning an analytical technique into high throughput screening of chiral compounds. Physicochemical properties and enantioseparation capability of polymeric β-D-glucopyranoside based surfactants with different head groups and chain lengths were compared. Moreover, the comparison of polymeric α- and β-D-glucopyranoside 4,6-hydrogen phosphate, sodium salt were further explored with regard to enantioseparations of ephedrine alkaloids and b-blockers. The concept of multiplex chiral MEKC-MS for high throughput quantitation is demonstrated for the first time in scientific literature.

Processos biotecnológicos dependem significativamente das técnicas de separação e purificação utilizadas, para manter boa relação custo-benefício na produção em escala industrial de produtos biotecnológicos com fins comerciais, industriais e terapêuticos. A aplicação do sistema micelar de duas fases aquosas (SMDFA) é proposta como alternativa para purificação de biomoléculas/biopartículas, pois permite sua separação e análise, muitas vezes sem que essas percam sua atividade ou propriedades desejadas. Com essa técnica é possível realizar uma partição seletiva que possibilita altos rendimentos. Esse trabalho destinou-se a estudar o emprego dessa metodologia na extração e purificação de Adenovírus em sistema micelar de duas fases aquosas formado por Triton X-114/Suspensão viral. Os ensaios foram realizados em sistemas de 5 g seguindo um planejamento fatorial completo (23) com 4 pontos centrais. Os fatores estudados foram temperatura de extração, pH da suspensão viral e concentração do tensoativo. Sistemas contendo massas de 3g, 10g e 40g foram avaliados. Foi avaliado o efeito do processo de extração com SMDFA sobre a integridade e infectividade de Adenovírus. Alguns dos parâmetros avaliados no processo foram a recuperação da potência viral (RPv) e a recuperação da potência viral específica (RPvø). Esses dois parâmetros avaliam a inativação do Adenovirus pelo processo de extração e ambos apresentaram melhoras quando comparados com a própria suspensão viral para alguns dos sistemas estudados (i.e RPv:341 % e RPvø 1466 %). Esses resultados indicam que o SMDFA foi capaz de particionar seletivamente as partículas virais infecciosas. De acordo com os resultados do planejamento é possível aumentar ainda mais esses resultados controlando as variáveis concentração de tensoativo, pH da suspensão viral e temperatura de extração. / Biotechnological processes depend significantly on separation and purification techniques used to maintain cost-effective industrial-scale production of biotechnological products for commercial, industrial and therapeutic uses. The application of the aqueous two-phase micelar system (ATPMS) is proposed as an alternative for purification, since it allows the separation and analysis of biomolecules /bioparticles, often without loses of activity or their properties. This allows to perform a selective partition that enables high yields. This work aims to study the use of this methodology in the extraction and purification of adenovirus in micelle aqueous two-phase formed by TritonX-114/Viral suspension. All assays were performed in 5 g systems following a full factorial design (23) with four central points. The studied factors were extraction temperature, pH of the viral suspension and concentration of the surfactant. Systems containing masses of 3g, 10g and 40g were evaluated. Extraction procedure effects over integrity and infectivity of adenovirus were also evaluated. Some of the parameters evaluated in the viral recovery process were viral potency (RPv) and recovery of viral specific potency (RPvø). These two parameters measure the inactivation of Adenovirus by the extraction process and both showed improvement when compared with the viral suspension for some of the systems studied (i.e RPv: 341% and RPvø 1466%). These results show that ATPMS selectively partition the infectious viral particles. According to the results of the experimental design is possible to increase, even further, these results controlling the surfactant concentration, viral suspension pH and temperature of extraction.

Étude du comportement d'ADN en solution et aux interfaces : actuellement, l'objectif de parvenir à une plus grande efficacité dans les processus de compaction de l'ADN, dans l'innovation des capteurs d'ADN et dans l'étude des changements dans les propriétés interfaciales générés entre les surfaces métalliques et les molécules d'ADN, est devenue un grand domaine d'intérêt en bioingénierie. Cette section de la thèse propose le couplage des techniques rhéométriques, électrochimiques et optiques afin d'effectuer une étude détaillée du comportement de molécules d'ADN en solution et aux interfaces, en fonction de la température, de la concentration en ADN et du potentiel électrique. Tout d'abord, le comportement rhéologique des solutions d'ADN, ainsi que la détection des concentrations critiques (C* et Ce), est discuté à partir d’expériences de rhéométrie en cisaillement permanent et harmonique. Après avoir étudié les propriétés des solutions d'ADN, des techniques électrochimiques et optiques ont été utilisés pour identifier les changements structurels aux interfaces Au/ADN et Pt/ADN, ainsi que pour décrire l'arrangement des chaînes d'ADN dans la double couche électrochimique pour les régimes dilué et semi-dilués. La réponse obtenue par Spectroscopie d'Impédance Électrochimique (EIS), Modulation Interfaciale de la Capacitance (MIC) et Résonance des Plasmons de Surface (SPR) reflète un processus d'adsorption des molécules d'ADN sur les surfaces métalliques. Finalement, en utilisant des concentrations d’ADN dans le régime dilué, on a étudié la formation de nanoparticules de chitosane-ADN avec stœchiométrie définie pour le transfert de gènes.Le rôle de la dynamique micellaire et la rhéologie dans la libération contrôlée de médicaments: le transfert ciblé d'ingrédients actifs (vectorisation) est un grand défi pour la recherche thérapeutique. Ce procédé est utilisé pour contrôler le transfert des protéines, des gènes et des médicaments vers une cellule cible en l'associant à un vecteur. Les molécules pour la chimiothérapie sont souvent hydrophobes et ont besoin d’un vecteur pour être transférées. Dans cette section de la thèse, on cherche à comprendre les dynamiques d'échange collectives (fusion et fission) entre micelles de copolymères triblocs amphiphiles à l'équilibre et hors équilibre. Ensuite, on étudie les dynamiques d'échange collectives entre ces micelles, choisies comme vecteurs, et des liposomes, choisis comme cellules modèles. On utilise une technique de fluorescence avec un dérivé de pyrène hydrophobe pour suivre les processus de fusion et de fission. Après avoir caractérisé la structure des copolymères amphiphile et avoir étudié leur dynamique à l'équilibre et hors l'équilibre, nous proposons une technique de fluorescence qui permet de quantifier les dynamiques collectives de vectorisation entre les micelles et les liposomes. Les effets de la variation de la concentration de liposomes et de l’adsorption du chitosane sur la membrane du liposome et sur les micelles ont été étudiés. / Nowadays, the target for reaching a greater efficiency in DNA compaction processes, the innovation ofDNA sensors development and the study of changes in the interfacial properties generated between metalsurfaces and DNA molecules has become an area of great interest in bioengineering. This section of thethesis proposes the coupling of rheological, electrochemical and optical techniques to perform a detailedstudy of DNA molecules behavior in the bulk state of the solution and at the interface with two differentmetallic surfaces, as a function of parameters such as temperature, DNA concentration and electricpotential. Firstly, the rheological behavior of DNA/buffer solutions, as well as the evidence of the criticalconcentrations (C★ and Ce) is discussed from simple steady state and oscillatory dynamic shearexperiments. After studying DNA solutions properties, electrochemical and optical techniques are used toidentify structural changes in Au/DNA and Pt/DNA interfaces and to describe the arrangement of DNAchains in the electrochemical double-layer as a function of concentration and within each characteristicregime, i.e. dilute and semi-dilute regimes. The obtained response trough Electrochemical ImpedanceSpectroscospy (EIS), Modulation Interfacial of the Capacitance (MIC) and Surface Plasmon Resonance(SPR) techniques reflects an adsorption process of DNA molecules taking place onto the metal surfaces.Finally, by selecting DNA concentrations in the dilute regime, we studied the formation of chitosan-DNAnanoparticles with defined stoichiometry for gene transfer.The specific delivery of active ingredients, known as vectorization, has actually become a greatchallenge in therapeutic research. This process has been used to control the distribution of activeingredients such as proteins, genes for gene therapy and drugs, to a target by associating it with avector. Molecules for chemotherapy are frequently hydrophobic and require vectorization to betransported to the target cell. In this section of the thesis, we look up to understand the collectiveexchange dynamics (fusion and fission) between amphiphilic block copolymer micelles at the equilibriumand out of the equilibrium, and the exchange dynamics between these micelles (representing vectors)and the simplest model of cells (liposomes). We used a fluorescent technique with hydrophobic pyrenederivative to probe the fusion and fission of micelles at equilibrium. After characterizing amphiphilicblock copolymers structure and studying their dynamics in and out of equilibrium, we proposed a timescan fluorescence technique to quantify the collective vectorization dynamics between amphiphilic blockcopolymer micelles and liposomes. The effect of the variation of several parameters such as liposomeconcentration and a chitosan adsorption were investigated in order to control the vectorizationdynamics between these vectors and cells models.