O gênero Passifloraceae possui mais de 400 espécies, sendo uma delas Passiflora incarnata L., detentora de diversas aplicações , principalmente no âmbito farmacêutico como uma planta capaz de atuar no sistema nervoso central para o tratamento de distúrbios da ansiedade. As substâncias mais abundantes existentes nas folhas dessa espécie pertencem à classe dos flavonóides, principalmente os C-glicosilados, que podem estar diretamente relacionados à atividade farmacológica da planta. Assim, o objetivo do presente trabalho , foi avaliar a influência (ou não ) de fatores externos na concentração dos flavonóides, em um cultivo experimental realizado na região de Botucatu .Os flavonóides analisados foram a luteolina, homoorientina, vitexina, isovitexina e orientina , com sua quantificação realizada por HPLC-UV/DAD e expressos em flavonóides totais calculados com base no padrão de rutina, através do método descrito por Pereira (2002). Realizou-se a correlação quimiométrica destes resultados pela análise multivariada , através de PCA (análise de componentes principais ) e HCA (análise hierárquica de agrupamentos), para avaliar se ocorreu a influência direta de determinados elementos do solo (pH, macronutrientes e micronutrientes) e do ambiente (temperatura e umidade) no teor de flavonóides encontrados nas folhas. Concluiu-se que as amostras se apresentaram de forma heterogênea no decorrer do cultivo (quanto à classificação ) , porém que alguns minerais presentes no solo (por exemplo, ferro, boro e cobre ), possuem relação inversa à concentração dos flavonóides totais encontrados nas folhas de P. incarnata. / Passifloraceae genus has more than 400 species and one of them is Passiflora incarnata L., which holds different application, mainly on the pharmaceutical scope as an herb capable to act on the central nervous system to the treatment of anxiety disturbs. The more copious compounds that exist on the leaves of this specie belong to flavonoid category, mainly C-glicosilates that could have direct relationship with pharmacologic properties of the herb. So, the objective of this task was to evaluate concentration in experimental growth made on Botucatu?s place. Analysed flavonoids were luteolin, homoorientin, vitexin, isovitexin and orientin, which were quantified by HPLC-UV/DAD and expressed as total flavonoids as rutin, through described method from Pereira (2002). Results were analysed through chemometric´s correlation, with PCA (principal components analysis) and HCA (hierarchic clusters analysis), to evaluate if some soil members (pH, macro and micronutrients) and ambient members (temperature and humidity) affected or not the flavonoids content on the leaves. It was conclude that all samples appeared heterogeneous in the course of growth, but some members present on the soil (as example iron, cupper and bore), have opposite relationship with total flavonoids content on the leaves of P. incarnata.

O objetivo desse trabalho foi demonstrar a viabilidade do acoplamento entre as técnicas de cromatografia líquida de alta eficiência com a espectrometria de emissão óptica com plasma indutivamente acoplado (HPLC - ICP OES) em um equipamento da Thermo Fisher Scientific, visando análise de especiação elementar de espécies com maiores concentrações na amostra. A identificação de espécies moleculares associadas aos elementos Cu, Fe, Mg, e Zn em amostras de castanha de caju foi adotada como modelo para comprovar tal acoplamento. Para que o ICP OES pudesse adquirir sinais de emissão ao longo de um tempo determinado, ou seja, para que o equipamento pudesse coletar um sinal transiente, fundamental para o acoplamento HPLC - ICP OES, foi necessário executar algumas modificações. Para isso, foram feitas alterações na placa eletrônica principal do equipamento, e trocada sua caixa controladora de gases. Os parâmetros instrumentais do ICP OES foram otimizados em uma série de ensaios com o intuito de estabelecer e avaliar desempenho do instrumento e a robustez do método empregado nas análises de castanha de caju. Determinações das concentrações totais de Ca, Cu, Fe, Mg, Mn, Ni, e Zn foram feitas em amostras de castanha de caju e nos materiais de referência NIST 1515 e NIST 1547, apresentando uma exatidão do método aceitável, levando-se em consideração o preparo de amostra e a medida no ICP OES. As determinações totais elementares nas amostras de castanha de caju indicaram que os teores de Ca foram de (0,04 ± 0,01) % ; Cu (26,3 ± 5,0) mg kg-1; Fe (47,9 ± 8,8) mg kg-1; Mg (0,31 ± 0,04) %; Mn (25,6 ± 3,9) mg kg-1; Ni (24,8 ± 1,9) mg kg-1; Zn (66,3 ± 2,4) mg kg-1. Para identificação de espécies moleculares associadas aos elementos Cu, Fe, Mg, e Zn, foi feita nas amostras de castanha de caju uma extração de metais e proteínas utilizando-se de uma solução extrato de NaOH 1 mol L-1. O extrato ainda foi submetido à uma etapa de clean-up, com precipitação das proteínas com acetona e ressupenção das mesmas com tampão Tris-HCl, pH 7,4. As proteínas e metais ressupensos em tampão foram analisados no sistema HPLC ICP OES, em que a separação cromatográfica ocorreu através de uma coluna de exclusão por tamanho (SEC), a detecção molecular através do detector UV-Vis do HPLC, e a detecção elementar através do ICP OES. As espécies moleculares identificadas estão no intervalo de massa molecular compreendido entre 2 e 177 kDa, sendo que Cu, Fe e Zn apresentam associação com essas espécies moleculares. Foram monitorados ainda, no ICP OES, os sinais de intensidade de emissão de P e S. Esses elementos também apresentaram associação com espécies moleculares com peso molecular entre 2 e 177 kDa, sugerindo que essas espécies se tratam de metaloproteínas. / The aim of this study was to demonstrate the capability of coupling between liquid chromatography techniques of high efficiency with the optical emission spectrometry with inductively coupled plasma (HPLC - ICP OES) in an equipment of Thermo Fisher Scientific, aiming elemental speciation analysis of species with higher concentrations in the sample. The identification of molecular species associated with elements Cu, Fe, Mg and Zn in cashew nuts samples was adopted as a model to prove such coupling. For ICP OES could get emission signal through a certain time, so that the equipment could collect a transient signal, critical to the HPLC coupling - ICP OES, it was necessary to perform some modifications in the instrument. For this, changes were made to the main electronic board of the equipment and also changed their gas controller box. The instrumental parameters of the ICP OES were optimized in a series of tests in order to establish and evaluate performance of the instrument and the robustness of the method employed in cashew nut analysis. Determination of the total concentration of Ca, Cu, Fe, Mg, Mn, Ni and Zn were made in cashew sample and reference materials NIST 1515 and NIST in 1547, with an accuracy acceptable method, taking into account the sample preparation and measurement by ICP OES. The elemental total determinations in cashew nut samples indicated that Ca were (0.04 ± 0.01)%; Cu (26.3 ± 5.0) mg kg-1; Fe (47.9 ± 8.8) mg kg-1; Mg (0.31 ± 0.04)%; Mn (25.6 ± 3.9) mg kg-1; Ni (24.8 ± 1.9) mg kg-1; Zn (66.3 ± 2.4) mg kg-1. For the identification of molecular species associated with the elements Cu, Fe, Mg, and Zn was performed on samples of cashew nuts an extract of metals and proteins using an extract solution of NaOH 1 mol L-1. The extract was further subjected to one-step of clean-up with protein precipitation with acetone following by a resuspension with Tris-HCl buffer, pH 7.4. Proteins and metals resuspended in buffer, were analyzed in the HPLC - ICP OES system, wherein the chromatographic separation took place by a size exclusion column (SEC), the molecular detection by UV-Vis detector of HPLC, and elemental detection by ICP OES. The molecular species were identified in the molecular weight range between 2 and 177 kDa, and Cu, Fe and Zn are associated with these molecular species. Were also monitored in the ICP OES, P and S emission intensity signals. These elements also associated with molecular species having a molecular weight between 2 and 177 kDa, suggesting that this species are metalloproteins.

O aciclovir é um anti-viral usado mundialmente para o tratamento de herpes (do tipo HSV-1 e HSV-2). Acredita-se que o vírus da herpes está presente em cerca de 90% das pessoas em estado de latência. O tratamento principal em casos onde a doença se manifesta, lesões nos lábios e mucosas, é o aciclovir. No Brasil, para renovar ou fazer um novo registro de medicamento, a Agência Nacional de Vigilância Sanitária (ANVISA) exige testes que expõem o fármaco a ambientes extremos (ácido, base, luz, calor, umidade, oxidação) para gerar produtos de degradação que dependendo da concentração no produto acabado, devem ser submetidos a ensaios toxicológicos. O estudo de degradação forçada também permite elucidar a estabilidade intrínseca do fármaco, contribuindo para o entendimento do mecanismo de degradação da substância, o que, posteriormente, ajuda a compreender quais fatores físicos e químicos devem ser controlados para manutenção da estabilidade. Este trabalho tem objetivo de validar um método de análise para quantificação do aciclovir e seus produtos de degradação por cromatografia líquida de interação hidrofílica (Hydrophilic interaction chromatography - HILIC) com detecção por arranjo de diodos (DAD) como também degradar a amostra em diferentes ambientes. / Acyclovir is an anti-viral used worlwide for herpes treatment (HSV-1 and HSV-2 types). It is estimated that herpes virus is present in almost 90% of people in his latent state. Acylcovir is the main treatment in cases where virus expresses itself. In Brazil, to renew or to make a new registration of medicines, National Agency of Sanitary Surveillance (ANVISA) demands tests to expose the medicine to extreme conditions (acid, basic, light, heat, humidity, oxidation) in order to generate degradation products that, depending on the concentration of the row product, will undergoes toxicological assays. The forced degradation study also allow to elucidate the intrinsic stability of medicine, contributing to understand the degradation mechanism of the substance leading to help the understanding of which physical and chemical factors must be controlled to keep stability. The main objective of this work is to validate a method of analysis to quantify acyclovir and his degradation products using Hydrophilic Interaction Liquid Chromatography (HILIC) based on Diode Array detection as well as to degraded the sample in different conditions.

<p style="text-align: justify;">Betain je bioaktivno jedinjenje, koje ima značajne fiziolo&scaron;ke funkcije u čovekovom organizmu kao osmolit i donor metil grupa za mnoge biohemijske procese. Endogena sinteza betaina je uglavnom nedovoljna da zadovolji potrebe organizma te je zbog toga neophodan njegov unos preko hrane. Žitarice predstavljaju glavni izvor betaina u ljudskoj ishrani. Jedan od načina da se poveća unos betaina u ljudskoj ishrani je preko funkcionalne hrane obogaćene betainom. Prema regulativi evropskog udruženja cerealija za doručak (European Breakfast Cereal Association) iz 2012. god. (Commission Regulation (EU) No 432/2012) dozvoljeno je deklarisanje zdravstvene izjave za prehrambene proizvode koji sadrže najmanje 500 mg betaina po porciji (40g flips proizvoda) uz navođenje napomene da se zdravstveni efekti mogu očekivati ukoliko se dnevno unese 1500 mg betaina. Zdravstvena izjava glasi: &bdquo;betain doprinosi normalnom metabolizmu homocisteina&ldquo;. Povećano interesovanje za zdravu ishranu dovelo je do veće potražnje za alternativnim žitaricama, gde spelta zbog svojih biolo&scaron;kih, agronomskih, prehrambenih i medicinskih karakteristika kao i bogatih nutritivnih svojstava, zauzima značajno mesto. Glavni cilj ove teze je bila optimizacija ekstrudiranja i proizvodnje funkcionalnog flips proizvoda od speltinog integralnog bra&scaron;na sa dodatkom betaina na dvopužnom ekstruderu. Prvi put je u procesu prehrambenog ekstrudiranja primenjena analiza vi&scaron;eciljne optimizacije u kombinaciji sa neuronskim mrežama i genetskim algoritmom kako bi se dobio maksimalan sadržaj betaina u flips proizvodu pri minimalnoj potro&scaron;nji specifične mehaničke energije. Definisan je nov funkcionalni proizvod sa betainom koji bi potencijalno pro&scaron;irio asortiman flips proizvoda na trži&scaron;tu i utvrđene su njegove fizičke, funkcionalne, teksturalne, reolo&scaron;ke i senzorske osobine. Sadržaj betaina i potro&scaron;nja specifične mehaničke energije su ključni izlazi koji predstavljaju konkurentne ciljeve u procesu ekstrudiranja. Za proizvodnju flips proizvoda pobolj&scaron;anog nutritivnog kvaliteta i funkcionalnog profila koji sadrži od 1601,6 do 1764,7 mg betaina u 40 g (čime se i zadovoljava preporučeni dnevni unos betaina) i kome odgovara potro&scaron;nja specifične mehaničke energije od 97,4 do 114,1 Wh/kg optimalni su sledeći procesni parametri: protok materijala od 20,45 do 24,04 kg/h, vlaga materijala od 18,6 do 19,44 % i brzina obrtanja pužnice od 250 obrtaja u minuti. Da bi se dobili pouzdani rezultati za sadržaj betaina u spelti i flips proizvodima razvijena je, optimizovana i validovana metoda tečne hromatografije visokih perfomansi. Metoda je uspe&scaron;no primenjena i u analizi betaina u žitaricama, pseudožitaricama i njihovim proizvodima.</p> / <p>Betaine is a bioactive compound that has significant physiological functions in the human organism as an osmolite and methyl group donor for many biochemical processes. The endogenous synthesis of betaine is generally insufficient to satisfy the requirements of the organism, therefore its intake is necessary through the food. Cereals are the main sources of betaine in human nutrition. One of the ways to increase betaine intake in human nutrition is through betaine enriched functional foods. In the 2012 European Union Regulation (Commission Regulation (EU) No 432/2012), it is permissible to declare a health statement for foods containing at least 500 mg betaine per portion (40g of extruded product), stating that the health effects can be expected if 1500 mg of betaine are consumed on a daily basis. The health statement states: &quot;Betaine contributes to the normal metabolism of homocysteine.&quot; Increased interest in healthy food has led to the higher demand for alternative cereals, where spelt takes a significant place due to its biological, agronomic, nutritional and medical characteristics, as well as its rich nutritional properties. The main goal of this work was the optimization of extrusion and production of the functional snack product from spelt wholegrain flour with the addition of betaine which is performed on twin-screw extruder. According to our best knowledge, there was no investigation in which the multi-objective optimization (MOO) analysis coupled with artificial neural network and genetic algorithm was applied to the extrusion cooking process in order to achieve simultaneously maximum betaine content and minimum energy consumption in the snack spelt product. A new functional snack product that can potentially expand the range of snack products and its physical, functional, textural, sensory and rheological properties were defined. The betaine content and specific mechanic energy are the key outputs that represent competing objectives in the extrusion proces. To produce snack product with improved nutritive quality and functional profile which contains betaine content from 1601,6 to 1764,7 mg/40g, which would contribute to the recommended daily intake of betaine, correspond to specific mechanical energy from 97,4 to 114,1 Wh/kg, following optimal parameters were obtained: feed rate from 20,45 to 24,04 kg/h, moisture content from 18,6 to 19,44 % and screw speed at 250 rpm. Also, optimization, development and validation of the high pressure liquid chromatographic method for measuring betaine content in spelt and snack products was also designed and applied.</p>

O presente trabalho teve como ênfase avaliar a atividade herbicida de aleloquímicos em extratos aquosos de sementes da Canavalia ensiformis (feijão-de-porco) através de ensaios de alelopatia e determinação de compostos fenólicos com atividade alelopática, utilizando a técnica de cromatografia líquida de alta eficiência (HPLC). Com este objetivo realizaram-se bioensaios de germinação, medida do crescimento da radícula e produção de biomassa em placas de Petri, usando como substrato solo e papel e como planta alvo as ervas daninhas corda-de-viola (Ipomoea sp.) e trapoeraba (Commelinna sp). Para os ensaios de alelopatia foram preparados extratos aquosos de sementes de Canavalia ensiformis em diferentes concentrações. Os bioensaios mostraram que os aleloquímicos presentes no extrato de feijão-de-porco quando aplicado em solo, têm um efeito fitotóxico menor do que aqueles bioensaios realizados em papel. Os resultados apontam que a espécie estudada apresenta potencial alelopático. Dentre os metabólitos secundários responsáveis pela atividade alelopática observada nos extratos desta espécie, estão os compostos fenólicos: ácido clorogênico, ácido ferúlico, ácido p-anísico e genisteína. As concentrações determinadas para os compostos fenólicos foram de 13,78 mg L-1 para o ácido clorogênico, 3,75 mg L-1 para o ácido ferúlico, 35,34 mg L-1 para o ácido p-anísico e 7,83 mg L-1 para a genisteína. Esses compostos apresentaram-se como fortes inibidores de germinação. Esta investigação mostra que na presença de solo os aleloquímicos presentes nesta leguminosa possivelmente diminuem sua atividade fitotóxica. / The objective of the present work was to study the herbicide activity of the allelochemicals in aqueous extracts from Canavalia ensiformis seeds through the allelopathy assays and phenolic compounds determination with allelopathi c activity, by High Performance Liquid Chromatography (HPLC). With this goal, germination bioassays, radicle length and biomass production in Petri plates were done, using paper and soil as substrate and the weeds species Ipomoea grandifolia and Commelinna benghalensis as target plants. For the allelopathy assays aqueous extracts from Canavalia ensiformis seeds were prepared in different concentrations. The bioassays showed that the allelochemicals present in the Canavalia ensiformis seeds, has a less phytotoxic effect compared to the bioassays done in paper. The results show that the studied specie has allelopathi c potential. Among the secondary metabolites responsibles for that allelopathic activity in this specie, are founded the phenolic compounds: chlorogenic acid, ferulic acid, p-anisic acid and genistein. The concentrations for the phenoli c compounds were determined: 13,78 mg L-1 for chlorogenic acid, 3,75 mg L-1 for ferulic acid, 35,34 mg L-1 for p-anisic acid and 7,83 mg L-1 for genistein. These compounds present themselves as strong germination inhibitors. This study shows that the allelochemicals present in the Canavalia ensiformis seeds have a lesser phytotoxic activity in soil.

Dinoflagelados foram os primeiros microorganismos reconhecidos como fonte primária de certos metabólitos, também conhecidos como ficotoxinas. Em geral são compostos não protéicos, de baixo peso molecular (250-3500 Da), apresentam estruturas químicas e modos de ação bem diferenciados. Envolvidos em florações, mortandade de peixes e intoxicações de seres humanos, devido ao consumo de mariscos contaminados, tornaram alvo de diversas pesquisas. Atividade antifúngica, hemolítica e citotoxicidade contra células tumorais são alguns exemplos da variedade de compostos e modos de ação que podemos encontrar. Como exemplo do potencial de aplicação, a pectenotoxina é candidata a agente quimioterápico e a maitotoxina é um estimulante da síntese e secreção do fator de crescimento. Em alguns casos, estudos com compostos de dinoflagelados alcançam níveis de patente. O risco à saúde pública e as perdas econômicas causadas pelas florações levaram ao estudo do ciclo de vida e interações ecológicas, taxonomia e aspectos moleculares das microalgas envolvidas nesses eventos. Também levou a criação de programas de monitoramento que levam em consideração observações ambientais, amostras de fitoplâncton, mariscos ou peixes, análise das amostras, avaliação e divulgação dos resultados e planos de ação em caso de eventos tóxicos. Considerando os estudos já realizados com dinoflagelados, estudamos algumas espécies isoladas do litoral norte do Estado de São Paulo sob aspectos biológicos, moleculares, químicos e monitoramento. A abordagem biológica revelou a presença de compostos capazes de inibir a proliferação celular e de alterar de alguma forma o citoesqueleto das células tumorais T47D nos extratos polares de A. fraterculus, P. gracile e P. mexicanum. Observamos também um possível potencial genotóxico das espécies A. fraterculus e P. gracile. Os aspectos moleculares, biológicos e químicos foram utilizados como ferramenta para o estudo de uma espécie de Ostreopsis cf ovata. A biologia molecular associada a morfologia nos levou a identificar duas cepas, mostrando que são geneticamente idênticas entre si e entre os espécimes isolados do Rio de Janeiro e Mediterrâneo e diferente daqueles de origem asiática, contribuindo para a compreensão da taxonomia, biogeografia e filogenia deste gênero. Ambas as cepas mostraram atividade hemolítica, mas as frações obtidas por cromatografia líquida de alta eficiência não tiveram a hemólise inibida por ouabaína. A análise por espectrometria de massa também não foi conclusiva devido à falta de padrão da toxina estudada. Portanto, sugerimos que os compostos hemolíticos presentes nas amostras sejam distintos da palitoxina e de seus análogos. No entanto, analisando os cromatogramas de espectrometria de massa, verificamos que a atividade biológica encontrada para as nossas amostras estão relacionadas a um conjunto de compostos. O programa de monitoramento piloto na maricultura da Praia da Cocanha em Caraguatatuba, litoral Norte do Estado de São Paulo foi realizado durante um ano. As análises de toxinas paralisantes (PSP) em bioensaios com camundongos e espectrometria de massa não detectaram toxinas nos mexilhões em concentrações que poderiam provocar intoxicação ao homem. Mas confirmamos o alerta quanto à presença de espécies potencialmente tóxicas no ambiente durante o período de estudo. Os aspectos abordados mostram como os compostos produzidos por dinoflagelados podem interferir de alguma forma na vida humana. Por um lado encontramos as finalidades farmacológicas na busca de novas drogas por outro o risco à saúde através do consumo de mariscos contaminados. Em ambos os casos, a continuidade das pesquisas trará muitos benefícios ao Homem. / Dinoflagellates were the firs microorganisms recognized as a primary source of certain metabolites, also known as phycotoxins. In general, compounds are non proteic, with low molecular weight (250-3500 Da), and have different chemical structures and modes of action. Frequently involved in algal blooms, killing fish and poisoning of humans due to consumption of contaminated seafood, they became the target of several researchers. Antifungal, hemolytic and cytotoxic activities against tumor cells are representative of the variety and modes of action that can be found. As examples of potential applications, the pectenotoxin is considered promising candidate for cancer chemotherapy; and the maitotoxin is described as stimulant of the synthesis and secretion of growth factor. In some cases, dinoflagellates compounds reached patent levels. The need to protect human health and fisheries industry led to studies of life cycle, ecologic interactions and taxonomic and molecular aspects of microalgae involved in harmful blooms. These events led to the establishment of monitoring programs to detect phytotoxins and toxic microalgae in mariculture farms worldwide. Based on researches carried out with others dinoflagellate, we studied some species isolated in the North coast of the state of São Paulo, on biological, molecular, chemical and monitoring aspects. Biologic assays indicated the presence of compounds in the dinoflagellates extracts that are able to inhibit cell proliferation and to induce changes in T47D cells cytoskeleton. We suggest a genotoxic potential for the compounds in the polar extracts of A. fraterculus and P. gracile. Biologic, molecular and chemical aspects were used to study two strains of Ostreopsis cf ovata. Molecular biology showed that both are genetically identical to O. ovata isolated from the Mediterranean Sea and Rio de Janeiro, but different from O. ovata collected in Asian waters. This result allowed a better understanding of genetic diversity and the recognition of more reliable (stable) morphological characters. Using mouse erythrocytes assay, hemolytic activity was detected in both strains of O. cf ovata. However, the hemolytic component was not inhibited by ouabain. Analyses by mass spectrometry of polar extract were inconclusive due to the lack of toxins standards. Therefore, we suggest that the compounds in the samples are distinct from palytoxin. Mass spectrometry results indicate that the biological activity of our samples can be related to several substances. The monitoring program in the mussel farm at Cocanha Beach, Caraguatatuba, was carried during one year. Mouse bioassays for paralytic shellfish poisoning toxins (PSP) performed in mussels showed that concentrations were below regulatory levels for human consumption. These low levels of PSPs were also confirmed by mass spectrometry. However, potentially toxic species were detected, and a long-term monitoring program is proposed and highly recommended. Research shows that compounds produced by dinoflagellates can interfere in human life both in positive and negative ways. On one side is the search for new drugs for pharmacological purposes; on the other hand the health risk represented by the consumption of contaminated seafood. In both cases, the continuity of the research will bring many benefits to humans.

Une échelle de taille contenant 20 substituants usuels en chimie a été construite à partir des barrières de rotations de N-aryl-thiazoline-2-thiones. L’énergie d’activation ΔG≠rot qui est mesurée reflète l’encombrement stérique du substituant en ortho du cycle benzénique. Les perturbations électroniques, et les facteurs externes tels que la température ou le solvant sont négligeables. La grande sensibilité du modèle proposé conduit au classement suivant Me > Cl et CN > OMe > OH. Ces classements divergents décrits dans la littérature seront discutés. Une limitation du modèle proposé concerne les substituants très volumineux comme le CF3 et l’iPr qui apparaissent plus petit qu’ils ne le sont en réalité à cause d’un encombrement stérique dans l’état fondamental qui abaissent la valeur de la barrière de rotation. / A steric scale of 20 recurrent groups was established from comparison of rotational barriers on N-(o-substituted-aryl)-thiazoline-2-thione atropisomers. The resulting energy of activation ΔG≠rot reflects the spatial requirement of the ortho substituent borne by the aryl moiety, electronic aspects and external parameters (temperature and solvent) generating negligible contributions. Concerning divergent rankings reported in literature, the great sensitivity of this model allowed to show unambiguously that a methyl appears bigger than a chlorine, and gave the following order in size: CN > OMe > OH. For the very bulky CF3 and iPr groups, constraints in the ground state decreased the expected ΔG≠rot values resulting in a minimization of their apparent sizes.

Chitooligosaccharides are composed of glycosamin and N-acetylglycisamin residues. Gel permeations chromatography is employed for the separation of oligomers, cation exchange chromatography is used for the separation of homologes and isomers. Trideuterioacetylation of the chitooligosaccharides followed by MALDI-TOF mass spectrometry allowes for the quantitation of mixtures of homologes. vMALDI LTQ multiple-stage MS is employed for quantitative sequencing of complex mixtures of heterochitooligosaccharides. Pure homologes and isomers are applied to biological assays. Chitooligosaccahrides form high-affinity non-covalent complexes with HC gp-39 (human cartilage glycoprotein of 39 kDa). The affinity of the chitooligosaccharides depends on DP, FA and the sequence of glycosamin and N-acetylglycosamin moieties. (+)nanoESI Q TOF MS/MS is used for identification of a high-affinity binding chitooligosaccharide of a non-covalent chitinase B - chitooligosaccharide complex. DADAA is identified as the heterochitoisomer binding with highest affinity and biostability to HC gp-39. Fluorescence based enzyme assays confirm the results. / Chitooligosaccharide sind aus Glycosamin und N-Acetylglycosamun aufgebaut. Gelpermeationschromatographie wird für die Trennung von Oligomeren verwendet, die Kationenaustauschchromatographie wird zur Trennung von homologen- und Isomerengemischen angewendet. Trideuterioacetylierung der Chitooligosaccharide gefolgt von einer Analyse mittels MALDI-TOF MS erlaubt die quantitative Analyse von Homologengemischen. vMALDI LTQ multiple-stage MS wird angewendet zur Sequenzanalyse und Quantifizierung komplexer Gemische von Heterochitooligosacchariden. Reine Homologe und Isomere werden für biologische Assays verwendet. Dabei zeigt sich, dass Chitooligosaccharide mit HC gp-39 hochaffine Komplexe bilden. Die Affinität der Chitooligosaccharide hängt vom DP, FA und der Sequenz der Chitooligosaccharide ab. (+)nanoESI Q TOF MS/MS wird erfolgreich angewendet zur Identifizierung eines Chitooligosaccharides, das mit hoher Affinität an Chitinase B (Serratia marcescens) bindet. DADAA wurde als die Sequenz des Isomers identifiziert, das mit höchster Affinität und Biostabilität an aktive Chitinase B bindet. Fluoreszenz basierte Enzymassays konnten dieses Ergebnis bestätigen.

In order to increase the analysis frequency and thereby achieve a better understanding of the kinetics and dynamics of the chemical process without increasing the workload of the already strained analytical laboratory at Cambrex Karlskoga AB, this projects goal was to investigate whether a crude prototype for mobile on-line HPLC-analysis with automatic sampling and dilution could be built based on certain flow-injection analysis techniques. It was possible to achieve dilution with good repeatability even though saturation effects in the filter proved problematic. Separation and dilution of a binary mixture was also successful as proof-ofconcept.

Knowledge about selenium content of foods containing selenium species is very important in terms of both nutrition and toxicity. Bioavailability of selenium species for human body is different from each other. Hence, speciation of selenium is more important than total selenium determination. In the selenium speciation study, chicken breast samples, selenium supplement tablets and egg samples were analyzed for their selenium contents. In chicken breast study, chickens were randomly categorized into three groups including the control group (25 chickens), inorganic selenium fed group (25 chickens) and organic selenium fed group (25 chickens). After the optimization of all the analytical parameters used throughout the study, selenomethionine, selenocystine, Se(IV) and Se(VI) were determined using Cation Exchange-HPLC-ICP-MS system. In selenium supplement tablet study, anion and cation exchange chromatographies were used to determine selenium species. Arsenic is known as toxic element, and toxicity of inorganic arsenic species, As(III) and As(V), is much higher than organic arsenic species like arsenobetaine and arsenosugars. Hence, speciation of arsenic species in any matrix related with human health is very important. In the arsenic speciation study, Cation Exchange-HPLC-ICP-MS and Cation Exchange-HPLC-ES-MS systems were used to determine arsenobetaine content of DORM-2, DORM-3 and DOLT-4 as CRMs. All of the parameters in extraction, separation and detection steps were optimized. Standard addition method was applied to samples to eliminate or minimize the matrix interference. Thiols play an important role in metabolism and cellular homeostasis. Hence, determination of thiol compounds in biological matrices has been of interest by scientists. In the thiol study, Reverse Phase-HPLC-ICP-MS and Reverse Phase-HPLC-ES-MS systems were used for the separation and detection of thiols. For the thiol determination, thiols containing &ndash / S-S- bond were reduced using dithiothreitol (DTT). Reduction efficiencies for species of interest were found to be around 100%. Reduced and free thiols were derivatized before introduction on the column by p-hydroxymercuribenzoate (PHMB) and then separated from each other by using a C8 column. In the real sample measurement, yeast samples were analyzed using HPLC-ES-MS system.