No presente trabalho foi avaliada a capacidade antioxidante in vitro dos extratos do cotilédone da semente de girassol rajada, obtidos por extração seqüencial com solventes de diferentes polaridades, bem como avaliado o potencial antioxidante in vitro do extrato que apresentou maior capacidade in vitro. Todos os parâmetros in vitro (sistema β-caroteno/ácido Iinoléico, métodos FRAP, DPPH, ORAC e Rancimat) indicaram o extrato aquoso (EAq) como o de maior capacidade. Neste extrato, o ácido clorogênico (12,88%) foi identificado como o principal componente dos ácidos fenólicos. Na avaliação da capacidade antioxidante in vitro, ambas as determinações empregadas (TBARS e perfil de ácidos graxos) indicaram o EAq como capaz de exercer um efeito protetor sobre os ácidos graxos poliinsaturados dos tecidos adiposo, cerebral, hepático e plasmático de ratos Wistar machos recém-desmamados. / The aims of this work were to evaluate the in vitro antioxidant capacity of listed sunflower cotyledon extracts, obtained by a sequential extraction with solvents of different polarities, and to evaluate the in vitro antioxidant potential of the sample extract with highest in vitro capacity. Ali the in vitro parameters (β-carotene/linoleic acid system, FRAP, OPPH, ORAC and Rancimat methods) indicated the aqueous extract (EAq) as the extract with highest capacity. In this extract, the chlorogenic acid (12.88%) was identified as the principal fraction of phenolic acids. In the in vitro antioxidant capacity evaluation, both determinations used (TBARS and fatty acids profile) gave indication that the EAq was capable to exerce a protective effect on the polyunsaturated fatty acids of the adipose, cerebral, hepatic and plasm tissues of Wistar male rats just-weaned.

Ce travail de thèse s’inscrit dans un contexte industriel. Il consiste en la caractérisation de lots bruts de SG1 : le N-tert-butyl-N- (1-diethylphosphono-2,2-dimethylpropyl)-N-oxyle. La spécificité de cette espèce est son caractère radicalaire stable. La première partie de ce travail consistait à développer une méthode d’analyse, facilement applicable en laboratoire d’analyse de contrôle sur site industriel pour la caractérisation de la pureté en SG1 dans des lots synthétisés. La méthode choisie a été développée par chromatographie liquide haute performance (HPLC). Pour ce faire, un étalon de SG1 a été préparé par purification de SG1 brut industriel en utilisant la technique séparative chromatographique de partage centrifuge (CPC). L’identification des impuretés suspectées majoritaires dans les lots de SG1 bruts a ensuite été réalisée par spectroscopie RMN ou par l’étude des fragments obtenus par spectrométrie de masse en tandem par ionisation electrospray. A l’aide de ces techniques, quinze impuretés majori- taires ont été identifiées. Des méthodes de quantification ont ensuite été développées pour douze de ces impuretés par diverses techniques analytiques chromatographiques : par HPLC-UV, HPLC-MS ou GC-MS ou par des techniques spectroscopiques Infra-Rouge ou encore par conductimétrie. / This PhD study has been carried out within an industrial context. It deals with the characterization of industrial batches of N-tert-butyl-N- (1-diethylphosphono-2,2-dimethylpropyl)-N-oxyle also called SG1. The distinctive feature of this molecule lies in its specific stability which is uncommon for radical species. The first part of this study consists in developping an analytical method, easy to implement in a QC laboratory for the determination of SG1 purity. The chosen analytical method has been developped by High Performances Liquid Chromatography (HPLC). This method requires the use of a standard sample of SG1, which has been obtained after purification of industrial batches of SG1 by the use of an innovative separative technology, the Centrifugal Partition Chromatography (CPC). The identification of the impurities, suspected as the major impurities in the industrial batches of SG1, has been carried out by NMR spectroscopy or by the study of their fragmentation observed by Tandem Mass Spectrometry – Electrospray Ionization. The use of these techniques enabled the identification of fifteen major impurities. Quantification methods by various techniques (HPLC-UV, HPLC-MS, GC-MS, other direct spectroscopic techniques like Infra-Red or other types of techniques like conductimetry) have been developped for twelve impurities.

Abstract Liquid chromatography (LC) combined with mass spectrometry (MS) is one of the most widely used techniques in modern analytical laboratories. Remarkable developments during the two previous decades in both techniques has made LC-MS the method of choice in various environmental, pharmaceutical and biochemical laboratories due to selectivity, sensitivity and versatility. The main focuses in this study were to develop new LC-MS methods to identify and quantify phenolic secondary metabolites in bilberry, lingonberry and hybrid bilberry, to study the biosynthesis of the main secondary metabolites (hypericin and hyperforin and their derivatives) in St John’s wort (SJW) both in vitro and in vivo (in plant), to identify in vitro metabolites of hyperforin in human liver microsomes and to identify the cytochrome P450 (CYP) enzymes responsible for their formation. Both high-performance liquid chromatography (HPLC) and ultra high-performance liquid chromatography (U-HPLC) in combination with time-of-flight (TOF) and triple quadrupole (QqQ) mass spectrometry were used in this study. Identification of 52 phenolic compounds from the leaves of bilberry, lingonberry and hybrid bilberry was accomplished. In total, seven of the identified compounds were reported for the first time in Vaccinium plants and several other compounds were reported for the first time in the studied plants. Incorporation of valine and isoleucine into acyl side chains of phloroglucinols (hyperforin and adhyperforin) via biosynthesis in shoot cultures of SJW was confirmed by using isotopically labeled amino acids and HPLC-MS/MS. Also, 29 biosynthetic in vitro products for HpPKS2 enzyme originating from SJW were identified based on accurate mass data. The metabolism of hyperforin was studied in human liver microsomes (HLM) for the first time. 57 metabolites for hyperforin were identified in the incubations with HLMs, using a substrate concentration of 1 μM. The phase I metabolism of hyperforin was suggested to rely mainly on CYP3A4 and on CYP2C family. / Tiivistelmä Nestekromatografia (LC) yhdistettynä massaspektrometriaan (MS) on yksi eniten käytetyistä analyysimenetelmistä nykyaikaisissa analytiikkalaboratorioissa. Viimeisten parin vuosikymmenen aikana LC-MS -laitteet ovat kehittyneet merkittävästi, ja nykyään LC-MS onkin paras menetelmä moniin ympäristö-, lääkeaine- ja biokemiallisiin laboratorioihin sen selektiivisyyden, herkkyyden ja monipuolisuuden vuoksi. Tässä väitöskirjassa kehitettiin uusia LC-MS –menetelmiä mustikan, puolukan ja mustikkapuolukan fenolisten sekundäärimetaboliittien tunnistamiseksi ja kvantitoimiseksi, mäkikuisman pääasiallisten sekundäärimetaboliittien (hyperisiini, hyperforiini ja niiden johdannaiset) tutkimiseksi in vitro- ja kasvinäytteistä sekä hyperforiinin aineenvaihduntatuotteiden tunnistamiseksi ja niitä muodostavien sytokromi P450 (CYP) entsyymien tunnistamiseksi ihmisen maksamikrosomeissa in vitro -menetelmin. Tässä työssä käytettiin sekä korkean erotuskyvyn nestekromatografia (HPLC) että ultra-korkean erotuskyvyn nestekromatografia (U-HPLC) yhdistettynä lentoaikamassaspektrometriin (TOF-MS) ja kolmoiskvadrupolimassaspektrometriin (QqQ-MS). Mustikan, puolukan ja mustikkapuolukan lehdistä tunnistettiin yhteensä 52 fenolista yhdistettä. Seitsemää näistä tunnistetuista yhdisteistä ei oltu aiemmin löydetty Vaccinium -suvun kasveista ja useat muut yhdisteistä löydettiin ensimmäistä kertaa nyt tutkituista kasveista. Valiinin ja isoleusiinin liittyminen floroglusinolien (hyperforiini ja adhyperforiini) asyylisivuketjuihin biosynteesin välityksellä varmistettiin isotoppileimattujen aminohappojen ja HPLC-MS/MS –mittausten avulla. Tässä työssä tunnistettiin myös 29 mäkikuismasta peräisin olevan HpPKS2 -entsyymin in vitro biosynteesituotetta tarkan massan mittausten avulla. Hyperforiinin metaboliaa tutkittiin ensimmäistä kertaa ihmisen maksamikrosomeissa (HLM). Hyperforiinille tunnistettiin yhteensä 57 aineenvaihduntatuotetta ihmisen maksamikrosomi-inkubaatioissa, kun hyperforiinin alkukonsentraatio oli 1 μM. Tämän tutkimuksen tulosten perusteella hyperforiinin faasi I metabolia tapahtuu pääasiassa CYP3A4:n ja CYP2C-perheen välityksellä.

Les constructeurs aéronautiques envisagent les systèmes de propulsion à hélices contra rotatives comme une alternative aux turboréacteurs, afin de réduire la consommation de carburant et les émissions des gaz à effet de serre. En raison de l’absence de carénage, la réduction du bruit engendrée par de tels systèmes représente un enjeu majeur pour les industriels. En particulier, le bruit de raies dû à l’impact des sillages de l’hélice amont sur l’hélice aval constitue une part significative de l’émission acoustique. Le travail présenté dans cette thèse a abouti à une méthode semi-analytique de prédiction de ce bruit d’interaction, intégrant de façon relativement réaliste les effets tridimensionnels des sillages de l’hélice amont et de la géométrie des pales de l’hélice aval. L’espace balayé par une pale est décomposé en tranches annulaires, déroulées pour décrire localement l’interaction en coordonnées cartésiennes. Le segment de pale obtenu est approché par un trapèze plat de forme et d’orientation quelconques. Une double stratégie est proposée pour la description du sillage. Premièrement, il peut être décrit par un modèle analytique tenant compte du vrillage et de l’expansion avec la distance au bord de fuite. Deuxièmement, il peut être post-traité à partir des calculs numériques. Ensuite, dans chaque tranche le déficit de vitesse ressenti par le segment de pale fait l’objet d’une décomposition de Fourier à deux nombres d’onde. Le calcul de la réponse aérodynamique instationnaire du segment est fait dans le domaine fréquentiel. Il étend des solutions analytiques existantes valables pour un segment rectangulaire, et prend en compte la compressibilité du fluide et la non-compacité des pales. On restitue ainsi les effets de la flèche, du vrillage et de la variation de la corde en envergure. Les fluctuations de portance induites sur les différents segments, obtenues par le calcul, sont utilisées pour construire une répartition de sources acoustiques équivalentes sur la surface réelle des pales, au sens de l’analogie acoustique. Le bruit en champ lointain est alors calculé en utilisant le formalisme de Ffowcs Williams & Hawkings, adapté au cas d’un dipôle tournant dans un écoulement uniforme. La méthodologie proposée a été implémentée dans l’outil ORION et évaluée avec des résultats numériques et des mesures en soufflerie. / Counter-rotating open rotors are seen as a possible alternative to turbofan engines for future subsonic aircraft propulsion, essentially for their higher fuel-efficiency. This technology leads to fuel saving sand to reduced green-house gas emissions. However, these benefits are balanced by some inherent draw-backs, as the increased noise radiation. Particularly, the tonal noise produced by the impingement of the wakes issuing from the front rotor onto the rear-rotor blades is recognized as a major contributor to the emitted noise. The research presented in this thesis led to a semi-analytical methodology to predict the rotor-rotor interaction tonal noise, including three-dimensional features of both rear-rotor blades and front-rotor wakes. The space is cut into annular regions, subsequently unwrapped for formulating the problem in equivalent Cartesian coordinates. Also, the obtained blade segments are assimilated as a set of flat trapezoids with arbitrary orientation, accounting for blade sweep and chord variations in the span wise direction. A double strategy is proposed for the description of front-rotor wakes. First, an analytical model is proposed in which wake direction and diffusion are deduced from the blade stagger angle and axial distance between the rotors. Secondly, a strategy for post-processing numerical wakes is presented. In both cases, the oncoming excitation is expanded in a series of sinusoidal gusts with two aerodynamic wavenumber components. Using this information the unsteady loading on the rear-rotor blades is obtained, in the frequency domain, from an extension of Amiet’s theory for gust-air foil interaction to account for air foil sweep and chord variations, flow compressibility and source non-compactness. The obtained noise source is back-projected on the blade mean-camber surface. An extended far-field formulation is then used to predict the noise. This theory is derived in detail from Ffowcs Williams &Hawkings’ formalism adapted for acoustic dipoles rotating in a uniformly moving atmosphere. The pro-posed methodology has been implemented in the tool ORION and assessed by comparing its results with numerical simulations and wind-tunnel measurements.

The aim of this work is to identify the possibilities of the complementary usage of two analytical methods of data analysis, OLAP analysis and data mining represented by GUHA association rule mining. The usage of these two methods in the context of proposed scenarios on one dataset presumes a synergistic effect, surpassing the knowledge acquired by these two methods independently. This is the main contribution of the work. Another contribution is the original use of GUHA association rules where the mining is performed on aggregated data. In their abilities, GUHA association rules outperform classic association rules referred to the literature. The experiments on real data demonstrate the finding of unusual trends in data that would be very difficult to acquire using standard methods of OLAP analysis, the time consuming manual browsing of an OLAP cube. On the other hand, the actual use of association rules loses a general overview of data. It is possible to declare that these two methods complement each other very well. The part of the solution is also usage of LMCL scripting language that automates selected parts of the data mining process. The proposed recommender system would shield the user from association rules, thereby enabling common analysts ignorant of the association rules to use their possibilities. The thesis combines quantitative and qualitative research. Quantitative research is represented by experiments on a real dataset, proposal of a recommender system and implementation of the selected parts of the association rules mining process by LISp-Miner Control Language. Qualitative research is represented by structured interviews with selected experts from the fields of data mining and business intelligence who confirm the meaningfulness of the proposed methods.

[pt] A presente pesquisa de mestrado relaciona-se ao desenvolvimento e à validação de um método analítico para detecção de substâncias proibidas em matrizes biológicas de cavalos de corrida e seu impacto na confiabilidade metrológica de serviços laboratoriais voltados à atividade turfística visando (i) a especificação de desempenho de um laboratório antidoping capacitado para identificar a presença de substâncias proibidas acordadas pela indústria de carreiras de cavalo; (ii) a adequação às condições brasileiras dos métodos analíticos aplicáveis à determinação antidoping em matrizes biológicas de animais em competição e (iii) a definição de um padrão de excelência harmonizado aos critérios da International Federation of Horseracing Authorities entendidos como instrumentos para a conquista do credenciamento do laboratório antidoping e subseqüente participação em acordos internacionais de reconhecimento mútuo. Para fundamentar o desenvolvimento da pesquisa, o trabalho considerou como estudo de caso o Laboratório Antidoping do Jockey Club Brasileiro (LAD/JCB). Original no seu escopo, o trabalho contribui para ampliar a abrangência da atividade de acreditação laboratorial no Brasil tendo em vista tratar-se de uma nova categoria de laboratório de ensaio ainda não existente na matriz laboratorial credenciada cuja confiabilidade laboratorial foi desenvolvida à luz dos critérios da norma internacional ISO/IEC 17025. No contexto do desenvolvimento são ainda discutidos aspectos relevantes relacionados ao sistema da gestão laboratorial; expressão das incertezas de medição associadas à determinação antidoping em consonância a práticas e procedimentos internacionalmente consensados; capacitação de pessoal técnico especializado; rastreabilidade ao Sistema Internacional de Unidades (SI) e validação e adequação de metodologias analíticas utilizadas em laboratórios antidoping. Atenção particular é direcionada ao Methode Alcalins Sur C- 18, desenvolvido pelo Laboratoire LAB-Contrôle antidopage da França, aplicável a determinação da presença de substâncias dopantes em matrizes biológicas (urina) de cavalos de corrida, utilizado como método fonte para o desenvolvimento de um método alternativo adequado às condições laboratoriais do Laboratório Antidoping do Jockey Club Brasileiro e moldado à detecção de substâncias dopantes mais freqüentes na atividade turfística brasileira. Tendo a cafeína sido estatisticamente identificada a substância mais presente nas análises realizadas pelo Laboratório Antidoping do Jockey Club Brasileiro ao longo de seus sete anos de atuação, o Methode Alcalins Sur C-18 foi adequado e validado para sua determinação analítica. À pesquisa desenvolvida atribuem-se, também, créditos pela introdução de um novo atributo na validação qualitativa da metodologia, denominado valor- limite de determinação, subsidiando a determinação de uma faixa de concentração, para que resultados incorretos (falso-positivos e falso- negativos) não constem de laudos emitidos. Adicionalmente à adequação e validação do método analítico desenvolvido, a pesquisa de mestrado contribuiu não apenas para atribuir visibilidade a uma nova categoria de laboratórios, mas, principalmente, para preparar o Laboratório Antidoping do Jockey Club Brasileiro para demonstrar sua competência técnica como estratégia de conquista de um credenciamento internacionalmente reconhecido. A presente pesquisa de mestrado voltada ao equacionamento de um problema inédito no Brasil de interesse da indústria relacionada à atividade turfística desenvolveu-se no contexto do Convênio FINEP/MCT no. 22.01.0692.00, Referência 1974/01, que aportou recursos do Fundo Setorial Verde Amarelo para direcionar o esforço de pesquisa em metrologia para a solução de dez projetos-piloto de interesse da indústria. No c / [en] This Master s Degree dissertation addresses the development in adjustment and validation and the metrological reliability of laboratory services covering horseracing activities. The work includes (i) the specification of the performance of an antidoping laboratory capable of identifying the presence of forbidden substances, as defined by the horse-racing industry; (ii) the validation of analytical methods applicable in doping tests of biological matrices of competing animals to Brazilian laboratories conditions, and (iii) the definition of a standard of excellence in harmony with the criteria of the International Federation of Horseracing Authorities as the tools utilized for the accreditation of the antidoping laboratory and the subsequent participation in international agreements of mutual recognition. In order to validate the methodology, the Brazilian Jockey Club Antidoping Laboratory (LAD/JCB) was used as a studycase. Original in its scope, this work contributes towards the widening of the accreditation of laboratories in Brazil, considering the fact this new category of testing laboratories is still nonexistent within the Brazilian network of accredited testing laboratories which considers the international standard ISO/IEC 17025 as the basic criteria. With regard to development, some relevant aspects are still being discussed in relation to the laboratory management system: the unreliability of the measurements for the determination of doping, in view of internationally accepted practices and procedures; qualification of specialized technical personnel; traceability of SI (the International Unit System) and the validation and adjustment of analytical methodologies utilized in antidoping laboratories. Special attention is dedicated to Methode Alcalins Sur C- 18, developed by Laboratoire LAB-Contrôle Antidopage in France, applicable to the determination of the presence of doping substances in the biological matrix (urine) of racehorses, used as method source for the development of an alternative method adjusted to the Antidoping Laboratory of the Brazilian Jockey Club conditions and molded to detection of the more frequents doping substances in the brazilian horserancing activities. Having the caffeine been the doping substance statistically identified as the most frequent in the tests carried out by the Brazilian Jockey Club Antidoping Laboratory, along its seven years of services, the Methode Alcalins Sur C-18 was adapted and validated for its analytical determinations. The applied research carried out was also credited with the introduction of a new attribute to the methodology qualitative validation, designated as limit- value for determination, which provides key-information for the determination of a concentration band, in order that incorrect results (false-positive and false- negative) do not appear in the laboratory reports usually issued. Besides the adaptation and validation of the analytical method thus developed, this Master s Degree dissertation contributes not only towards greater visibility of a new category of laboratories, but mainly to the preparation of the Brazilian Jockey Club Antidoping Laboratory to demonstrate its technical capacity as a policy and strategy for becoming in the near future an internationally recognized accredited laboratory. The present M.Sc. Dissertation addresses the metrological reliability of antidoping laboratories, a pioneering issue in Brazil, aiming at the solution of a problem related to the horse-racing industry. It was developed under the financial support of the FINEP/MCT Agreement no. 22.01.0692.00, Reference 1974/01 (Green-Yellow governmental Sectoral Fund) which droved research cooperative efforts in metrology to the solution of ten pilot-projects. In this case specifically, cooperated to the development of the M.Sc. research entitled Metrological reliability of the Antidoping Laboratory of the Brazilian Jockey Club

No description available.

<p>Therapeutic proteins are often lyophilized with excipients such as sucrose or trehalose to protect them during manufacturing and achieve a longer shelf-life. Formulation design for therapeutic proteins has been a trial-and-error process, and the mechanisms responsible for the stabilizing effects of excipients are not fully understood. Two proposed theories have been widely accepted: the water replacement theory and the vitrification theory.^1,2The water replacement theory suggests that excipients stabilize protein molecules in the solid state by forming hydrogen bonds that “replace” the hydrogen bonds to water that stabilize the protein in solution, while the vitrification theory asserts that proteins are stabilized by a glassy solid matrix of low mobility and does not require direct interactions between excipient and protein. A better understanding of the interactions between proteins and other components of the lyophilized matrix can facilitate rational formulation design and shorten the time in development. However, most of the analytical methods available can only provide information on the bulk properties of the lyophilized matrix such as moisture content and glass transition temperature (<i>T</i>_g); it has been difficult to measure the interactions between protein and excipient directly, if they exist. In order to characterize the interactions between protein and excipients in a lyophilized matrix with high resolution, a photolytic labeling method was developed in this dissertation, building on previous work in our research group. Photolytic labeling has long been used to identify protein-protein interactions <i>in vivo</i>.^3,4Common types of photo-reaction reagents and their applications are summarized in Chapter 1. The research described in this dissertation utilizes the diazirine functional group, which is activated after UV exposure and undergoes a free radical reaction to form covalent bonds with nearby molecules. The reaction can be used to identify the interactions between excipients and protein or peptide in a solid formulation. Previous studies in our lab have shown that photo-reaction can be applied to lyophilized solids to study protein-matrix properties and interactions in the solid.^5,6This dissertation seeks to further identify photo-reaction products and analyze them in a more quantitative way. </p><p> </p><p>Chapter 2 describes a quantitative analysis of photo-reaction products in solution and lyophilized solids using a model peptide, KLQ (Ac-QELHKLQ-NHCH₃). The purpose of the work in this chapter is to establish a quantitative analytical method for photo-reaction products, enabling studies of peptide-excipient interactions in lyophilized solids. KLQ was derivatized with a bifunctional probe NHS-diazirine (succinimidyl 4,4’-azipentanoate; SDA) at Lys5 to be photo-reactive. The SDA derivatized KLQ (KLQ-SDA) was used to study the photo-reaction products and examine excipient interactions. Identification and quantitation of photo-reaction products of KLQ-SDA was achieved with liquid chromatography mass spectrometry (LC-MS) and reversed phase HPLC (rp-HPLC). Important reaction products such as peptide-excipient adducts and peptide water adducts varied in different formulations. Unexpected reaction products such as unproductive “dead-end” products and peptide-phosphate adducts from buffer salt were also detected and quantified. Together, the photo-reaction products reflected the local environment near Lys5 of the peptide in the solid state. This study has provided a better understanding of photo-reaction with diazirine in the lyophilized solids together with a quantitative description of the local environment near Lys5. </p><p> </p><p>In Chapter 3, the photo-reaction products in lyophilized solids exposed to increasing moisture were analyzed, and the effect of increasing moisture on the local environment near the peptide was examined. Using the analytical method developed in Chapter 2, these studies explored whether peptide-water interactions, as measured by the formation of water adducts formed by photolytic labeling, are linearly correlated with an increase in solid bulk moisture content. Formulations containing the KLQ-SDA peptide were exposed to various relative humidity conditions and photolytic labeling was induced. Solids containing disaccharide excipients behaved differently from those containing amino acids when exposed to the same relative humidity condition, showing different levels of peptide-excipient and peptide-water adducts. With increasing moisture content in the solids, the formation of photo-reaction products did not mimic the pattern of solutions with same composition, indicating differences in the local environment. </p><p> </p><p>An alternative approach to studying lyophilized formulations using photolytic labeling is to incorporate photo-reactive excipients into the solid matrix. In Chapter 4, a new diazirine-labeled photo-excipient, photo-glucosamine (pGlcN), was chemically synthesized and incorporated into formulations of the therapeutic peptide salmon calcitonin (sCT) and compared with the commercially available diazirine-labeled amino acid, photo-leucine (pLeu). The studies in Chapter 4 further compared peptide-excipient interactions at the molecular level with two different photo-excipients, ionizable pLeu and unionizable pGlcN. Changing solution pH prior to lyophilization was expected to change ionic interactions between sCT and pLeu in the solid samples, resulting in different distributions of photo-reactions products; pH-dependent differences were not expected for pGlcN. The results demonstrated that the distribution of photo-reaction products varied with the composition of the formulation and the pH of the solution prior to lyophilization. The photo-reaction products in the pGlcN-containing formulation differed from those pLeu, showing a difference in the interactions of unionizable (pGlcN) and ionizable (pLeu) excipients with sCT in solid samples. </p><p> </p><p>The work in this dissertation has developed photolytic labeling as a tool to study lyophilized peptide formulations, and has provided a more quantitative understanding of the photo-reaction products that are produced from diazirine-labeled peptides or excipients in the solid state. A new photo-reactive excipient has also been presented (pGlcN), which showed different photo-reaction products than a commercially available photo-excipient (pLeu) and is promising for future study. Photolytic labeling for formulation development is still in its early stages, and additional research regarding reaction mechanism and complementary stability studies is needed. Nevertheless, the results presented in this dissertation support continued development of photolytic labeling as a practical method for formulation design and development. </p><p> </p>

Läsinlärning är en stor del av undervisningen i ett visst stadium. Lärare arbetar med olika läsinlärningsmetoder i undervisningen för att skapa möjlighet för elever att lära sig läsa. Syftet med denna studie är att ta reda på lärares uppfattningar om syntetiska och analytiska metoder för elever i förskoleklass och årskurs 1. Syftet besvaras genom följande frågeställningar: Vilka syntetiska och analytiska metoder använder lärare för att utveckla tidig läsinlärning i förskoleklass och årskurs 1? Hur använder lärare syntetiska och analytiska metoder för att utveckla elevernas tidiga läsinlärning? Hur uppfattar lärare att syntetiska och analytiska metoder kan bidra till en utveckling av elevernas läsinlärning? Studien genomfördes med en kvalitativ metod där data samlades in genom semistrukturerade intervjuer med sex lärare. Teorin som studien har utgått ifrån är sociokulturellt perspektiv. Resultatet redovisas under fem rubriker som är kopplade till studiens syfte: Lärarnas arbete med läsinlärning, den tidiga läsundervisningen, det fortsatta arbetet med läsinlärning, elevers synliga utveckling och elevers motivation och engagemang mot läsutveckling. Resultatet visar att lärarna uppskattar metoderna för elevernas läsinlärning. Lärarna använder modellerna Bornholm, Fonomix, ASL och dialogisk högläsning för att utveckla elevernas läsinlärning och hur de används redogörs i resultatet. Resultatet visar även att eleverna synligt utvecklas i sin läsinlärning och att lärarna uppfattar att motivation behövs för att elever ska komma vidare i sin läsinlärning. / The learning to read process is a major part of the education at a certain stage. Teachers work with different methods to teach students to read, and to create opportunities for reading development. The purpose with this study is to answer which perceptions teachers’ have on synthetic and analytical methods for students in preschool and grade 1. The purpose is answered through the following questions: Which synthetic and analytical methods do teachers use to develop early learning to read in preschool and grade 1? How do teachers use synthetic and analytical methods to develop students early reading? How do the teachers’ perceptions of synthetic and analytical methods contribute to students’ development in the learning to read process? The data was collected through semi-structured interviews with six teachers. The study is based on the sociocultural perspective. The result is reported through five headings that are linked to the purpose of this study: Teachers work with learning to read, the early reading instruction, the following work with learning to read, students’ visible development and students’ motivation and commitment to reading development. The results show that the teachers approve of the methods for students’ development for learning to read. The teachers use the models Bornholm, Fonomix, ASL and dialogical reading to develop students learning to read and how they use them is explained in the results. The results shows that teachers perceptions are that motivation also is needed for students’ development to read.

Att lära sig läsa är en väsentlig del i dagens skola och samhälle. Därför läggs mycket fokus på just läsundervisning i de tidiga åren. Lärares kunskap och didaktik har stor betydelse för att främja elevernas läsinlärning. Det krävs en förståelse för läsprocessens komplexitet och vilka förmågor hos eleverna som är viktiga att utveckla. Följaktligen är studiens syfte att undersöka hur sex lärare på tre olika skolor uppfattar att en god läsundervisning etableras. Undersökningen vill belysa lärarens syn på vilka förmågor som är viktiga att utveckla i den tidiga läsundervisningen, vilka läsinlärningsmetoder som används och hur. Studien tar ansats i den sociokulturella teorin, där den proximala utvecklingszonen stöttar inlärning. Läraren behöver ha läsinlärningskunskaper för att kunna ta reda på var eleven befinner sig i sin utveckling och för att stötta hen vidare till en högre nivå i kunskapstrappan. Undersökningen genomförs med individuella semistrukturerade intervjuer med samtliga lärare. De semistrukturerade intervjuerna används för att samtalet ska vara någorlunda öppet men ändå följa en struktur. Resultatet visade att läsmotivation, avkodning och språkförståelse behövs för en effektiv läsinlärning. I studien framträder flera läsinlärningsmetoder, tre syntetiska och en analytisk. De tre syntetiska metoderna fokuserar mer på avkodning medan den analytiska metoden börjar i förståelsen. Undersökningen visar att det är givande att arbeta med flera metoder eftersom man dels kan anpassa undervisningen på olika nivåer, dels för att de olika metoderna passar olika elever. Genom varierande arbetssätt och läsinlärningsmetoder gynnas fler elever av läsundervisningen, vilket möjliggör att fler barn lär sig läsa. / Learning to read is an essential part of today`s school and society. Therefore, a lot of focus is placed on reading teaching in the early years. Teachers´ knowledge and didactics are great importance in promoting students´ learning to read. It requires an understanding of the complexity of the reading process and which abilities of the student are important to develop. Consequently, the aim of the study is to investigate how six teachers at three different schools perceive that good reading teaching is established. The survey wants to shed light om the teacher`s view of which abilities are important for pupils to develop in early reading teaching, which reading learning methods are used and how. The study takes an approach in the sociocultural theory, where the proximal development zone supports learning. The teacher needs to have reading learning skills to be able to find out where the student is in his development and support him further to a higher level on the knowledge ladder. The survey is carried out with individual semi-structured interviews with all teachers. The semi-structured interviews are used so that the conversation is reasonably open but still follows a structure. The results showed that reading motivation, decoding and language comprehension are needed for effective reading learning. Several reading learning methods appear in the study, three synthetic and one analytic. The three synthetic methods focus more on decoding while the analytical one begins in understanding. The survey shows that it is rewarding to work with several methods, partly because you can adapt the teaching at different levels, partly because the different methods suit different students. By using more varied working methods and reading learning methods, the teaching of reading benefits more students, which enables more children to learn to read.