This thesis describes the synthesis and characterization of metal-encoded polystyrene microspheres with a narrow size distribution designed for mass cytometry-based immuno- and oligonucleotide-assays. These particles were prepared by multiple stage dispersion polymerization techniques using polyvinylpyrrolidone (PVP) as a steric stabilizer. As a cytometeric technique, mass cytometry necessitated metal-encoded microspheres to perform the same roles of fluorescent microspheres used in conventional flow cytometry. The first role of the microsphere was to be able to act as a platform (classifier microspheres) for bioassays. Secondly, the microspheres should be suitable for mass cytometry machine calibration as standards. To perform these roles, metal-encoded microspheres were required to have certain size, functionality and metal content criteria. Lanthanide elements were chosen as the metals for encoding the microspheres for their low natural abundance in biological systems and for their similar chemistry. My goal was to employ two-stage dispersion polymerization, of styrene in ethanol, to introduce the lanthanide salts along with excess acrylic acid in the second stage, one hour after the initiation. Acrylic acid deemed to serve as a ligand for the lanthanide ions, through its carbonyl group, so the lanthanide ions get incorporated into the microsphere while acrylic acid is copolymerizing with styrene. Using two-stage dispersion polymerization, I could synthesize lanthanide encoded microspheres with narrow size distribution and high lanthanide content. However the lanthanide content distributions were unexpectedly much broader than the size distribution obtained. In addition, I could not attach biomolecules to the surface of such particles. In an attempt to improve the characteristics of these microspheres, I employed modified versions of multiple stage dispersion polymerization and seeded emulsion polymerization to grow functional polymer shell on the surface of the particles prepared by dispersion polymerization. Moreover, I coated the lanthanide encoded microspheres with silica shell which enabled me to grow another layer of functional-silica. Consequently, I could use these particles as classifier microspheres for mass cytometry-based immunoassays as well as fluorescence-based oligonucleotide-assays.

Ce travail étudie le potentiel de l'électrophorèse capillaire couplée à la spectrométrie de masse à ionisation par plasma (ICP/MS) pour le criblage d'une bibliothèque de ligands en fonction de leur affinité pour l'europium en milieu hydro-organique. Cette méthode permet, d'une part, d'évaluer l'affinité des ligands phosphorés en moins de deux heures et en utilisant moins de 15 ng de ligand, et d'autre part, de déterminer les constantes de complexation. Les résultats sont en accord avec ceux obtenus par titrage spectrophotométrique.Parallèlement, une bibliothèque de copolymères pour l'extraction solide/liquide de l'europium a été étudiée. Le protocole d'extraction mis au point permet de les classer selon leur affinité pour celui-ci en milieu hydro-organique et en utilisant 60 mg de copolymère. Pour les plus prometteurs, les propriétés de reconnaissance et la sélectivité La3+/Eu3+/Lu3+ ont été évaluées.

Although the benefits of ICP-MS are well documented, the determination of analytes at low levels in high concentrations of matrix elements has proved difficult. As ICP-MS is a 'flow into' instrument the deposition of salts throughout the system is a common cause of significant loss of signal. The application of desolvation of aluminosilicate samples, to aid in the production of more robust plasma conditions, was investigated to increase the efficiency of the ICP in processing the sample. The performance of the ICP-MS was monitored with different cone arrangements and by running the skimmer cones at elevated temperatures. An alternative to modification of the instrument is to employ chemical modification of the sample and hence the separation of Au and Pt from an aluminosilicate matrix via the use of solid phase extraction (SPE) columns were investigated as a means of dealing with high levels of dissolved solids. OVB based SPE columns were found to give high retentions of Au and Pt when chelated with ammonium pyrrolidinediethylcarbamate (APOC). A second alternative that avoided digestion of the aluminosilicate matrices, was to carry out the analysis using laser ablation (LA). LA-ICP-MS is becoming increasingly used for trace elemental analysis but as yet no universal calibration method is available. The general problems associated with matrix matched standards are inherent as the ablation mechanism and plasma conditions can differ dramatically with very small changes in matrix composition. Hence the addition of chromophores was employed to increase the absorption of the laser energy. The use of vanillic, nicotinic and pyrazinoic acid were used to improve the ablation of pressed powder discs at the laser wavelength of 213 nm. Synthetic aluminosilicate discs and standard additions were both employed for the calibration and determination of Ti.

abstract: Applications of non-traditional stable isotope variations are moving beyond geosciences to biomedicine, made possible by advances in multiple collector inductively coupled plasma mass spectrometry (MC-ICP-MS) technology. Mass-dependent isotope variation can provide information about the sources of elements and the chemical reactions that they undergo. Iron and calcium isotope systematics in biomedicine are relatively unexplored but have great potential scientific interest due to their essential nature in metabolism. Iron, a crucial element in biology, fractionates during biochemically relevant reactions. To test the extent of this fractionation in an important reaction process, equilibrium iron isotope fractionation during organic ligand exchange was determined. The results show that iron fractionates during organic ligand exchange, and that isotope enrichment increases as a function of the difference in binding constants between ligands. Additionally, to create a mass balance model for iron in a whole organism, iron isotope compositions in a whole mouse and in individual mouse organs were measured. The results indicate that fractionation occurs during transfer between individual organs, and that the whole organism was isotopically light compared with food. These two experiments advance our ability to interpret stable iron isotopes in biomedicine. Previous research demonstrated that calcium isotope variations in urine can be used as an indicator of changes in net bone mineral balance. In order to measure calcium isotopes by MC-ICP-MS, a chemical purification method was developed to quantitatively separate calcium from other elements in a biological matrix. Subsequently, this method was used to evaluate if calcium isotopes respond when organisms are subjected to conditions known to induce bone loss: 1) Rhesus monkeys were given an estrogen-suppressing drug; 2) Human patients underwent extended bed rest. In both studies, there were rapid, detectable changes in calcium isotope compositions from baseline - verifying that calcium isotopes can be used to rapidly detect changes in bone mineral balance. By characterizing iron isotope fractionation in biologically relevant processes and by demonstrating that calcium isotopes vary rapidly in response to bone loss, this thesis represents an important step in utilizing these isotope systems as a diagnostic and mechanistic tool to study the metabolism of these elements in vivo. / Dissertation/Thesis / Ph.D. Chemistry 2011

The aim of the thesis was to determine whether the individual Se-speciation in the mammalian organism are affected by the form of received selenium. Selenium is an essential micronutrient important for humans and animals. It plays an important role in the antioxidant protection of the organism and in the conversion of thyroid hormones. In our experiment the laboratory Wistar rats were divided into three groups. Each group had a different diet. The rats were fed with selenium in the form of soy, sodium selenite and extracted rapeseed meal. Urine samples were regularly collected during the four-week experiment and in the end of the feeding study, the blood serum was also collected. The total selenium content was measured by ICP-MS, while the individual Se-speciation in urine and serum by HPLC coupled with ICP-MS. In the urine the identified speciation were methylselenocystein (MeSeCys), trimethylselenium (TMSe) and selenosugar 1 and 3. In the blood serum the measured speciation were TMSe, selenite, selenate and selenosugar 1. For the group fed with sodium selenite the measured values in the urine were generally higher, this might be due to a higher overall intake and also an inorganic form of selenium with a lower absorbency. Groups that received selenium from plant sources took in several Se-compounds and the total measured content of Se-speciation and secretion dynamics were not significantly different. Additionally speciation of selenosugar 2 was measured for the group fed with rapeseed meals, which in the other groups did not appear. When receiving selenium from plant sources the biotransformation in the mammalian organism differs in comparison to receiving selenium from mineral salts. The initial hypothesis that Se-speciation is influenced by the form of selenium administered in the diet was confirm by our results. Since the group fed rapeseed showed similar results as the group fed a standard feed with soy, the extracted rapeseed meal could serve as a good source in livestock nutrition.

O grande magmatismo intracontinental ocorrido a 133 Ma na Bacia do Paraná foi desenvolvido por um sistema fissural no qual a interação da Pluma Tristão da Cunha na base da litosfera (com ou sem contaminação crustal) gerou derrames, diques e sills no Brasil, nos Estados do RS, SC, PR, SP, MT, MS e GO, e em parte do Paraguai, Argentina e Uruguai. Na Formação Serra Geral, representativa deste intenso magmatismo, foram coletadas amostras de sills (RS, PR, SP e GO) cristalizados entre os sedimentos Paleozóicos pré-vulcânicos da Bacia do Paraná. Já haviam sido realizados diversos estudos geoquímicos a partir da década de 80 com objetivo de identificar as variações químicas de diques, derrames e sills da Formação Serra Geral. No entanto, a utilização de novas técnicas analíticas (EPMA e LA-ICP-MS), utilizadas no presente estudo, proporcionaram o entendimento das variações químicas de forma pontual em minerais ígneos. A geoquímica dos sills estudados caracteriza-os como sub-alcalinos toleíticos continentais que variam de basaltos a andesitos basáltico. Assim, podendo ser divididos em dois grupos químicos: sills do Rio Grande do Sul (baixo TiO2 < 2 wt.%, com concentrações inferiores de P2O5, Nb, Sr, Zr, Zn, Y e Pb e concentrações superiores de Rb, Th, U, e Cs) e sills do Paraná, São Paulo e Goiás (alto TiO2 >2 wt.%, de geoquímica oposta). Petrograficamente, os sills do RS possuem granulação mais fina, ocorrência restrita de cobre nativo e distribuição modal elevada para plagioclásios cálcicos, quando comparados ao sills do PR, SP e GO. A determinação geoquímica por micro-análise em plagioclásios e clinopiroxênios demonstra que as variações químicas identificadas em rocha total são relatas as modificações químicas ocorridas nestes minerais. Variações do coeficiente de partição (KD) de elementos traços compatíveis em diversas zonas de cristais de plagioclásio e clinopiroxênio possuem correlação na substituição dos elementos Ca, Na, Al, Fe e Mg nos sistemas NaAlSi3O8-CaAl2Si2O8 e Mg2Si2O6-CaMgSi2O6-CaFeSi2O6- Fe2Si2O6, respectivamente. Coeficientes de partição de cristais de plagioclásio e clinopiroxênio agora são determinados em diferentes porções de núcleo, intermédio e borda aprimorando o conhecimento da variação do KD até então determinado como um único valor apenas para o cristal. / The large intracontinental magmatism occurred at 133 Ma in the Paraná Basin was developed by a fissure system in which the interaction of Tristan da Cunha plume at the base of the lithosphere (with or without crustal contamination) caused lava flows, dikes and sills in the RS, SC, PR, SP, MT, MS and GO and part of Paraguay, Argentina and Uruguay. In the Serra Geral Formation, representative of intense magmatism, sills crystallized samples were collected (RS, PR, SP and GO) between the Paleozoic prevolcanic sediments of Paraná basin. Several geochemical studies have been carried out since the 80's in order to identify the chemical variations of dikes, sills and lava flows of the Serra Geral Formation. However, the use of new analytical techniques (EPMA and LA-ICP-MS) provided the study of chemical variations in a timely manner in igneous minerals. The geochemistry of the sills studied characterized them as sub-alkaline continental tholeiitic basalts ranging from the basalt to basaltic andesites. Divided into two chemical groups: sills of Rio Grande do Sul (low TiO2 <2 wt.%, With lower concentrations of P2O5, Nb, Sr, Zr, Zn, Y and Pb and higher concentrations of Rb, Th, U, and Cs) and sills of Parana, Sao Paulo and Goiás (high TiO2> 2 wt.%, geochemical opposite). Petrographically the sills of the RS have finer grain, restricted occurrence of native copper and modal higher to calcic plagioclase, compared to the sills of the PR, SP and GO. Geochemical determination by micro-analysis in plagioclases and clinopyroxenes show that the chemical variations identified in whole rock we describe the chemical changes occurring in these minerals. Variations of the partition coefficient (KD) of compatible trace elements in various parts of crystals of plagioclase and clinopyroxene were correlated in the replacement of Ca, Na, Al, Fe and Mg systems NaAlSi3O8-CaAl2Si2O8 and Mg2Si2O6-CaMgSi2O6-CaFeSi2O6-Fe2Si2O6, respectively. Partition coefficients of crystals of plagioclase and clinopyroxene are now determined in different portions of core, intermediate and rim enhance knowledge of the variation of KD previously determined as a single value only to the crystal.

This thesis deal with three individual Pagophilus groenlandicus canin teeth from the location Asva, Estonia dating from the Late Bronze Age (here 700-900 B.C). The aim of this thesis is to find out whether the three harp seal individual was local or non-local to Asva and further if they were migrants from the Atlantic Sea. The teeth enamel are analysed for strontium isotopes using LA-ICP-MS and the result was compared against the strontium 87Sr/86Sr values in various rivers in the Baltic Sea. This is to see how the harp seal has migrated around in the Baltic Sea. By placing the 87Sr/86Sr values against the rivers, it may seem that the three harp seals were non-local and not from the Atlantic Sea.

102f. / Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2013-03-27T17:17:03Z No. of bitstreams: 1 Tese doutorado final Reinaldo Barbour.pdf: 1224301 bytes, checksum: b37e38d9ef21943dba24e8f3638efbcd (MD5) / Approved for entry into archive by Ana Hilda Fonseca(anahilda@ufba.br) on 2013-04-19T13:16:36Z (GMT) No. of bitstreams: 1 Tese doutorado final Reinaldo Barbour.pdf: 1224301 bytes, checksum: b37e38d9ef21943dba24e8f3638efbcd (MD5) / Made available in DSpace on 2013-04-19T13:16:36Z (GMT). No. of bitstreams: 1 Tese doutorado final Reinaldo Barbour.pdf: 1224301 bytes, checksum: b37e38d9ef21943dba24e8f3638efbcd (MD5) Previous issue date: 2011 / CAPES / A determinação de impurezas em cobre eletrolítico, na faixa de μg g-1 e sub μg g-1, é muito importante para caracterizar o cobre de alta pureza (99,99%). As suas propriedades físicas e químicas como maleabilidade, condutividade térmica e elétrica são fundamentais nas aplicações industriais nos setores de eletricidade e eletrônica. Metais e ametais como Ag, As, Bi, Co, Cr, Fe, Mn, Ni, Pb, Sb, Se, Sn,Te e Zn modificam estas propriedades deteriorando sua maleabilidade e condutividade. A espectrometria de massas com plasma indutivamente acoplado (ICP-MS) tem sido muito empregada para a determinação elementar, em amostras industriais, devido às suas características, principalmente as relacionadas com a capacidade multielementar, a rapidez e a alta sensibilidade. As principais desvantagens da técnica são as interferências isobáricas e poliatômicas, além daquelas relacionadas à matriz. Este trabalho teve por objetivo estabelecer um procedimento analítico validado para a determinação de elementos contaminantes por ICP-MS em amostras de cobre eletrolítico, de forma a garantir os limites de quantificação exigidos para a certificação do cobre. O equipamento empregado foi operado nos modos padrão e CCT (célula de colisão que utiliza 1,0% (v/v) NH3 em He balanço) com comutação automática. As amostras e materiais de referência certificados foram digeridos, em triplicata, empregando bloco aquecedor, seguindo o procedimento: 10  1 mg de amostra com 2 mL de HNO3 1:1 (v/v) à 70ºC. A diluição final foi de 5000x, o que contribui para minimização dos efeitos de matriz provenientes da alta concentração dos íons de cobre (~200 ug g-1). Os isótopos selecionados no modo normal de operação do equipamento foram: 55Mn, 59Co, 60Ni, 68Zn, 75As, 107Ag, 118Sn, 121Sb, 128Te, 208Pb e 209Bi. Para o modo com célula de colisão, os isótopos foram: 52Cr, 56Fe, 57Fe, 77Se e 78Se. A faixa linear estudada foi de 0,01 a 2 ng g-1. Os limites de quantificação encontrados (μg g-1 no cobre) foram: Ag (0,05), As (0,2), Bi (0,05), Co (0,1), Cr (0,2), Fe (0,5), Mn (0,2), Ni (0,2), Pb (0,1), Sb (0,2), Se (0,2), Sn (0,2),Te (0,1) e Zn (0,5). A exatidão foi avaliada pela analise dos seguintes materiais de referência certificados: BAM-M381, BAM-M382, BAM-M383, BAM-M384 (Pure Copper, BAM; Federal Institute for Materials Research and Testing) e NIST 495, NIST 496, NIST 457 (National Institute of Standards and Technology) sendo que a maioria das recuperações obtidas foram entre 80 e 120% e os %RSD menores que 10% (Se < 20%). O efeito de matriz foi avaliado através das técnicas de calibração: calibração externa com padrões internos, calibração com adição de analito e calibração com uso dos CRMs e não houve diferença significativa dos resultados. A técnica DC-ARC OES foi empregada para comparação dos resultados. A avaliação dos resultados das amostras reais de bobinas foi feita por meio de ensaios interlaboratoriais e do critério z score. Por este critério, a maioria dos resultados obtidos foram considerados excelentes. Pode-se concluir que o método proposto, empregando ICP-MS, é adequado para determinação multielementar / Salvador

71f. / Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2013-04-09T13:15:43Z No. of bitstreams: 1 dissertação ALETE_definitiva_110330.pdf: 1412727 bytes, checksum: c409be4d388a7ed5944c1d0866387a9d (MD5) / Approved for entry into archive by Ana Hilda Fonseca(anahilda@ufba.br) on 2013-04-19T14:50:36Z (GMT) No. of bitstreams: 1 dissertação ALETE_definitiva_110330.pdf: 1412727 bytes, checksum: c409be4d388a7ed5944c1d0866387a9d (MD5) / Made available in DSpace on 2013-04-19T14:50:36Z (GMT). No. of bitstreams: 1 dissertação ALETE_definitiva_110330.pdf: 1412727 bytes, checksum: c409be4d388a7ed5944c1d0866387a9d (MD5) Previous issue date: 2010 / CAPES / O objetivo principal do trabalho foi à utilização da espectrometria de fluorescência de raios X com energia dispersiva (EDXRF) para determinação de elementos essenciais, tais como P, Cu, Fe, Mn e Zn em amostras de arroz na forma de pastilhas como método alternativo, obtendo resultados com precisão e exatidão aceitáveis no controle de qualidade dos alimentos. A validação foi feita comparando os resultados obtidos pelo método proposto com valores certificados e com aqueles obtidos utilizando decomposição das amostras em forno de micro-ondas com cavidade e os digeridos foram usados para determinação dos analitos por espectrometria de emissão óptica com plasma indutivamente acoplado (ICP OES) e espectrometria de massas com plasma indutivamente acoplado (ICP-MS). A precisão foi avaliada em termos da repetibilidade, sendo que foram preparadas onze pastilhas do material de referência certificado de farinha de arroz 1568a NIST e efetuadas três medições em cada pastilha obtendo-se estimativas do desvio padrão relativo inferiores a 5%. A exatidão foi verificada com o material de referência certificado (farinha de arroz 1568a NIST) e o t-teste pareado revelou que não havia diferença significativa entre os valores certificados e obtidos ao nível de confiança de 95%. Foram obtidos os seguintes limites de detecção (LODs): 0,037 g 100g-1 para P e 1,2; 3,9; 5,1 e 2,2 mg Kg-1 para Cu, Fe, Mn e Zn, respectivamente. O procedimento proposto foi aplicado para P, Mn e Zn e apresenta vantagens tais como a redução do tempo de análise e a eliminação da etapa de decomposição da amostra, porém apresenta a desvantagem dos maiores LODs. De modo geral, os resultados são novas informações sobre a composição mineral do arroz consumido na cidade de Salvador, Bahia, sendo úteis para formação de base de dados. / Salvador

O testemunho de firn IC-6 de 35,06 m de comprimento foi coletado no manto de gelo antártico (81°03’10,1”S e 79°50’09,1”W; 750 m de altitude) no verão austral de 2004/05. Este testemunho foi subamostrado usando um sistema de fusão contínua desenvolvido pela equipe do Climate Change Institute (University of Maine – Maine /EUA) em sala limpa (CLASSE 100). As 1380 amostras geradas foram analisadas em baixa, média e alta resolução no espectrômetro de massas Element 2 do CCI para 24 elementos traços (Sr, Cd, Cs, Ba, La, Ce, Pr, Pb, Bi, U, As, Li, Al, S, Ca, Ti, V, Cr, Mn, Fe, Co, Na, Mg e K). O testemunho representa 68 anos (1934 – 2002) de registro, segundo datação relativa baseada na variação sazonal nas concentrações dos elementos Na, Mg, Sr e Ca e dos íons Cl-, Na+ e Mg2+. A taxa de acumulação média para o local de amostragem é calculada em 0,30 m a-1 em Eq. H2O. As concentrações medidas foram consideradas baixas, dentro do esperado para o continente antártico e são similares a de outros estudos. As concentrações são controladas pelas variações climáticas sazonais (verão/inverno), pela distância de transporte e pelas fontes naturais e antrópicas desses aerossóis. Contribuições naturais de poeira continental e solo, oriundas principalmente da região de Patriot Hills, são as principais fontes para os elementos césio, bário, alumínio, titânio, vanádio, cromo, ferro, cobalto, manganês e terras raras. Os aerossóis marinhos, oriundos da superfície da cobertura de gelo marinho e transportados pelas massas de ar são fontes importantes de sódio, magnésio, estrôncio e enxofre. Os elementos lítio, cálcio e potássio apresentaram aportes consideráveis tanto de poeira continental como de aerossóis marinhos. Emissões vulcânicas globais e regionais (Monte Erebus e Ilha Deception) são consideradas importantes fontes de elementos traços. Elementos como vanádio, cromo, manganês, cobalto, bismuto, arsênio, cádmio e chumbo apresentaram uma significativa contribuição dessas fontes, variando desde 20% (chumbo e manganês) até 70% (cádmio e bismuto). Os fluxos de deposição natural de chumbo, cádmio, bismuto e arsênio, representam apenas uma pequena fração do total depositado na neve. Para esses elementos, as atividades antrópicas constituem o principal fator responsável por sua mobilização e transporte. / This dissertation examines a 35.06 m long firn core (IC-6) obtained in the Austral summer of 2004/2005 in the West Antarctic Sheet (at 81°03’10.1”S, 79°50’09.1”W; 750 m above sea level). This core was subsampled using a continuous melting system at the Climate Change Institute (CCI, University of Maine, Orono, Maine, USA) under a Class 100 room conditions. At CCI, 1380 samples went through an Element 2 mass spectrometer to determine (at low, middle and high resolutions) the concentration of 24 trace elements (Sr, Cd, Cs, Ba, La, Ce, Pr, Pb, Bi, U, As, Li, Al, S, Ca, Ti, V, Cr, Mn, Fe, Co, Na, Mg e K). The core records 38 years (1934 – 2002) of snow accumulation, as dated by seasonal variations of Na, Mg, Sr e Ca elements and Cl-, Na+ e Mg2+ ionic concentrations. The calculated mean annual accumulation is 0.30 m yr-1 (in H2O equivalent). Their mean concentrations are low and are similar to the ones found in other Antarctic ice core studies. Trace concentrations are controlled by seasonal variations (summer/winter), transport distance and by natural and anthropogenic sources. Cesium, barium, aluminium, titanium, vanadium, chrome, iron, cobalt, manganese and rare earths come from continental dust (mainly from the nearby Patriot Hills). Marine aerosols, from the pack ice surface and transported by air masses, are the sources for sodium, magnesium, strontium and sulphur. Both continental dust and marine aerosols contribute to the lithium, calcium and potassium concentrations. Global and regional (Mount Erebus and Deception Island) volcanic emissions are important trace elements sources, such as vanadium, chrome, manganese, cobalt, bismuth, arsenic, cadmium and lead, varying from 20% (lead and manganese) to 70% (cadmium e bismuth). Trace elements as lead, cadmium, bismuth e arsenic from natural sources have a minor contribution in the samples, human activities are thought to be the main cause for their mobilization and transport.