This thesis describes the design, construction, and evaluation of a series of LED array photolysis systems for high throughput photochemistry. Three generations of array systems of increasing sophistication are evaluated using calorimetric measurements and potassium tris(oxalato)ferrate(III) chemical actinometry. The results are analyzed using descriptive statistics and analysis of variance (ANOVA). The LEDs in the third generation array were shown to be statistically equivalent, with respect to light output, according to physical and chemical actinometry experiments. The third generation LED array was compared with a traditional 1000 W Xe arc lamp source in terms of cost, light intensity, and light stability. Two constant current drivers were evaluated with respect to LED array performance. The optimized third generation LED array was evaluated as the photolysis source for photochemical hydrogen production experiments using the supramolecular catalyst [{(bpy)2Ru(dpp)}2RhCl2](PF6)5. / Master of Science

Orientador: Lucia Regina Durrant / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-05T00:51:57Z (GMT). No. of bitstreams: 1 Kushida_MartaMitsui_D.pdf: 1944227 bytes, checksum: 5343a021abfd5cdca334152ade81151c (MD5) Previous issue date: 2005 / Resumo: Salmonella é uma das principais causas de doenças transmitidas por alimentos (DTAs) em todo o mundo, sendo objeto de preocupação dos principais organismos internacionais relacionados com a saúde pública. A análise deste patógeno faz parte da rotina de qualquer laboratório de controle de qualidade de alimentos e, embora exista uma grande variedade de métodos alternativos, a maioria desses laboratórios ainda utiliza o método cultural tradicional, caro, lento e trabalhoso. A principal barreira à disseminação dos métodos alternativos é o fato de serem presuntivos e, em caso positivo, exigirem o retorno ao método tradicional, para confirmação. Recentemente, entretanto, foi introduzida no Brasil uma técnica que não padece dessa desvantagem, o sistema BAX® de análise de Salmonella por reação de polimerase em cadeia (PCR). Além de ser confirmativo, o BAX® ainda é bem mais rápido, permitindo a obtenção dos resultados em 30h e com o mínimo de manipulação, pois é totalmente automatizado. A substituição do método tradicional pelo BAX® pode elevar significativamente a capacidade de análise dos laboratórios, porém, é um sistema novo, ainda pouco utilizado no país, sendo importante uma avaliação do seu desempenho na análise de produtos brasileiros, objetivo do presente trabalho. Para tanto realizou-se uma avaliação do Sistema BAX® em comparação com o método cultural tradicional, analisando 708 amostras de 22 categorias de alimentos em relação à presença de Salmonella pelos dois métodos, o que permitiu a determinação dos parâmetros de desempenho do sistema BAX®. Os resultados obtidos indicaram sensibilidade de 100%, especificidade de 98,6%, taxa de falsos positivos de 1,4% e taxa de falsos negativos de 0% / Abstract: Salmonella sp is one of most important food born microorganism in all the world, and it always has been focused among international public health organs. The food analysis for this pathogen is a subject of routine in all food quality control laboratories, and, besides there are several alternative methods for Salmonella sp detection, but most of the laboratories still work with the traditional technique of detection, which is very expensive and time consuming. The barrier to alternatives methods is the fact that most of them are considered presumptive, and need the traditional technique to confirm the positives results. An alternative technique that does not present the step of confirmation was recently, introduced in Brazil, the BAX¿ System for Salmonella sp, which works by polymerase chain reaction (PCR). It is a very quick detection method, safety, accurate which shows the analysis result in 30 hours, utilizing very few manipulation of samples, since is automatized. The substitution of traditional Salmonella sp detection method for BAX¿ system, can increase significantly the laboratories analysis capacity, but as it is a very new technique, is still unknown in our country. In despite of this, it is important to evaluate its performance in Brazilian food products, which is the aim of this research. A evaluation of BAX¿ system, compared with the Salmonell traditional method was then carried out, using 708 samples of 22 different food categories. The results showed sensibility of 100%, specificity of 98,6%, falsepositives ratio of 1,4% and false-negatives ratio of 0% for BAX¿ system / Doutorado / Doutor em Ciência de Alimentos

Submitted by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-09-21T18:40:15Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Gabriela Ribas Foresti.pdf: 1405427 bytes, checksum: a4af87ccdf8b530dff79a4594afdb7b4 (MD5) / Approved for entry into archive by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-09-21T18:40:28Z (GMT) No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Gabriela Ribas Foresti.pdf: 1405427 bytes, checksum: a4af87ccdf8b530dff79a4594afdb7b4 (MD5) / Made available in DSpace on 2016-09-21T18:40:28Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Gabriela Ribas Foresti.pdf: 1405427 bytes, checksum: a4af87ccdf8b530dff79a4594afdb7b4 (MD5) Previous issue date: 2015-06-26 / O enrofloxacino é um fármaco antibacteriano exclusivamente utilizado na clínica veterinária. Até o momento, não existem monografias oficiais para o controle de qualidade deste fármaco em suas formas farmacêuticas. No presente trabalho, métodos analíticos para determinação quantitativa do enrofloxacino contido em comprimidos palatáveis foram desenvolvidos e validados por: (a) espectrofotometria UV utilizando tampão fosfato de pH 7,4: etanol (95:5, v / v) como solvente, em 273 nm com faixa de concentração de 0,5 a 8,0 μg. mL-1. (b) CLAE com detecção detector de arranjo de diodos a 278 nm, fase móvel composta por tampão fosfato 0,04 M (pH 2,7 e 0,3% de trietilamina) e acetonitrila 75% (75:25, v/v), faixa de concentração de 20,0 a 80,0 μg mL-1 e tempo de retenção médio de 6 minutos; E (c) método de dissolução baseado em dados in vivo empregando-se 400ml de meio contendo HCl, pepsina e lecitina de soja, aparato pá, velocidade de 75 rpm e quantificação do fármaco por espectofotometria no UV visível. A validação de todos os métodos desenvolvidos foi realizada através da análise dos parâmetros: especificidade, linearidade, precisão, exatidão e robustez, conforme estabelecido para medicamentos de uso veterinário pelo VICH e todos os resultados obtidos confirmaram que os métodos desenvolvidos são úteis para o controle de qualidade de rotina de enrofloxacino em comprimidos palatáveis. / The enrofloxacin is an antibacterial drug used exclusively in veterinary clinic. To date, there are no official papers for the quality control of this drug in its pharmaceutical forms. In the present work, analytical methods for quantitative determination of enrofloxacin contained in palatable tablets were developed and validated by: (a) UV spectrophotometry using pH 7.4 phosphate buffer: ethanol (95: 5, v / v) as solvent, 273 nm with a concentration range from 0.5 to 8.0 ug. ml-1. (B) HPLC detection at 278 nm with a diode array detector, a mobile phase consisting of 0.04 M phosphate buffer (pH 2.7 and 0.3% of triethylamine) and 75% acetonitrile (75:25, v / v ), concentration range from 20.0 to 80.0 ug ml-1 and average retention time of 6 minutes; And (c) dissolution method based on in vivo data using up 400ml of medium containing HCl and pepsin soy lecithin, paddle apparatus, 75 rpm speed and quantification of the drug by UV visible spectrophotometry. Validation of all developed methods was performed by analysis of parameters: specificity, linearity, precision, accuracy and robustness, as provided for medicines for veterinary use by VICH and all results confirmed that the developed methods are useful for the control enrofloxacin routine quality in palatable tablets.

Orientadore: Isabel Cristina Sales Fontes Jardim, Sonia Claudia do Nascimento de Queiroz / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Quimica / Made available in DSpace on 2018-08-15T20:18:30Z (GMT). No. of bitstreams: 1 Morais_LaisSayuriRibeirode_D.pdf: 1161986 bytes, checksum: 39b7f874268f505aa38cca5ac6e52461 (MD5) Previous issue date: 2009 / Resumo: A região das nascentes do Rio Araguaia, na divisa dos estados de Goiás e Mato Grosso abriga uma porção das áreas de recarga do Aquifero Guarani, o qual é um dos maiores sistemas aquíferos do mundo e representa a principal fonte de água potável na região. Este reservatório pode estar comprometido pelo uso crescente de agrotóxicos nas plantações de soja e de milho da região. Diante deste cenário, neste trabalho foram desenvolvidos métodos analíticos para a determinação dos agrotóxicos imazetapir, nicossulfurom, imazaquim, carbofuram, atrazina, linurom, clorimurom-etil e diflubenzurom, utilizados em culturas de milho ou de soja. Para separação, identificação e quantificação dos agrotóxicos foram utilizadas a Cromatografia Líquida de Alta Eficiência com detecção por arranjo de diodos e a Cromatografia Líquida acoplada por Fonte de Ionização por Eletronebulização à Espectrometria de Massas em série. A extração em fase sólida (SPE), com cartuchos C18, foi selecionada e otimizada para a extração dos agrotóxicos em amostras de água. Para o preparo das amostras de solo foram estudadas as técnicas de extração por agitação mecânica, por banho ultrassônico e assistida por micro-ondas industrial e caseiro. Os métodos desenvolvidos foram validados obtendo-se limites de quantificação (LQ) em água, na faixa de 0,015-0,1 ng mL empregando SPE-HPLCDAD e 0,01 ng mL com SPE-LC-ESI-MS/MS. Com a extração assistida por micro-ondas caseiro e LC-ESI-MS/MS foram obtidos LQ de 1 ng mL para todos os agrotóxicos em solo. Em todos os métodos validados, os valores de recuperação dos agrotóxicos em um mesmo dia e em dias diferentes, ficaram dentro da faixa aceitável de 70-120 %. Os ensaios de repetitividade e de precisão intermediária para todos agrotóxicos apresentaram coeficientes de variação inferiores ao limite aceito de 15 %. Os métodos SPE-HPLC-DAD e SPE-LC-ESI-MS/MS foram aplicados em análises de amostras de água da região em estudo, verificando-se que a maioria dos agrotóxicos analisados encontrava-se abaixo do limite máximo de resíduos de 0,1 ng mL. Não foram detectados agrotóxicos nas amostras de solo coletadas em outras duas regiões com cultivo de milho e soja, empregando a extração assistida por micro-ondas caseiro e LC-ESIMS/ MS. Os parâmetros físico-químicos determinados para o imazetapir, imazaquim, nicossulfurom e clorimurom-etil mostraram que eles apresentam pouca afinidade pelo solo estudado, tendo grande disponibilidade para lixiviação, em conformidade com o índice de GUS / Abstract: The area near the Araguaia River, between Goiás and Mato Grosso States, is the location of a portion of the recharging of the Guarani Aquifer, which is one of the world¿s largest aquifer systems and an important source of drinking water. This reservoir could be threatened by the widespread use of pesticides in maize and soybean cultivation in this area. Thus, this work developed analytical methods for the determination of imazethapyr, nicosulfuron, imazaquin, carbofuran, atrazine, linuron, clorimuronethyl and diflubenzuron, pesticides used in maize and soybean cultivation. Pesticide separation, identification and quantification were performed using High-Performance Liquid Chromatography with Diode Array Detection (HPLC-DAD) and Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry (LC-ESI-MS/MS). Solid Phase Extraction (SPE) with C18 sorbents was optimized for sample extraction from water. Soil samples were extracted by mechanical shaking, sonication or microwave-assisted extraction with industrial and home microwave ovens. Methods were validated resulting in limits of quantification (LOQ) for the pesticides in water in the range of 0.015-0.1 ng mL, using SPE-HPLC-DAD, and 0.01 ng mL using LC-ESI-MS/MS. LOQ of 1 ng mL for all pesticides in soil were achieved using the home microwave oven and LC-ESI-MS/MS. Recoveries for pesticides with all methods were in the range 70-120 %. Relative standard deviations for repeatability and intermediate precision were less than 15 %. SPEHPLC- DAD and LC-ESI-MS/MS were employed for the analysis of samples of water from the recharge area and most of the pesticides were detected at concentrations below the minimum residue limit (MRL) of 0.1 ng mL established by the European Community. The home microwave oven and LC-ESI-MS/MS were used for the analysis of soil samples from two other regions of Brazil and the pesticides were not detected in these samples. Adsorption and desorption parameters were determined for imazethapyr, imazaquin, nicosulfuron and chlorimuron-ethyl, indicating that these pesticides have little affinity for the soil of the region of the Guarani Aquifer recharge, and show significant leaching potential, according to the ground water ubiquity score (GUS index) for these pesticides / Doutorado / Quimica Analitica / Doutor em Ciências

In the early 19th century a ground-breaking discovery was made that linked a dietary deficiency of a fat-soluble vitamin with the childhood disease known as rickets. The vitamin was named vitamin D and extensive research regarding the physiological importance of this vitamin followed ever since. It is currently known that vitamin D plays an important role in maintaining the calcium and phosphate homeostasis in the human body. Less clear evidence states the medical importance of vitamin D in the prevention and cancer, autoimmune disease and diabetes. Current literature shows that vitamin D has five distinct forms, vitamin D1 to D5, of which vitamin D2 and D3 are the most studied forms. The term “vitamin D” is often wrongfully used to include the vitamin D mother molecule, the vitamin D status indicator (25(OH)D), the biologically active form (1,25(OH)2D) and biologically inactive form (24,25(OH)2D). The interest for measurement of these vitamin D analogues is a continuously growing field both on individual and epidemiological level. For decades laboratories have struggled to produce a robust method capable of quantifying these different vitamin D analogues and uncovered a new form of complexity regarding the analysis of these analogues. The identification of the C3-epimeric forms of vitamin D metabolites has forced laboratories to rethink their analytical methods and several concerns were raised regarding the overestimation of the true vitamin D status by current analytical methods. The quantification of the biologically active and inactive forms of vitamin D is reported to be difficult and to date very few LC-MS/MS methods reported in the literature are able to quantify various vitamin D analogues. However, to our knowledge none of these methods are able to include the precursor vitamin D, the 25-hydroxylated metabolites, the biologically active and inactive metabolites, C3-epimers and isobaric compounds in a single run. Therefore the aim of this study was to develop, optimise and validate a LC-MS/MS method for the quantification of twelve vitamin D analogues in a single run. This was done by optimising the underlying LC-MS/MS parameters to ensure optimal analytical sensitivity in positive ESI mode and sufficient chromatographic separation between analytes with similar chemical properties. Furthermore, the optimised method was validated to ensure the accuracy and precision of the method before implementation into a clinical environment. The vitamin D analogues included in this study were vitamin D2, vitamin D3, 25(OH)D2, 25(OH)D3, 1,25(OH)2D2, 1,25(OH)2D3, 24,25(OH)2D2, 24,25(OH)2D3, 3-epi-25(OH)D2, 3-epi- 25(OH)D3, 7(OH)4C3 and 1α(OH)D3. A double liquid-liquid extraction with hexane and ethyl acetate were found to be the most efficient at extracting the vitamin D analogues from a serum matrix after matrix modification with sodium hydroxide. Recoveries of > 95 % (CV <10 %) were achieved for all the analytes. It was noted that a precursor adduct other than the molecular mass ion for a specific vitamin D analogue can produce a more abundant MS1 signal and that the ESI source parameters vary between analytes with different chemical properties and should therefore be optimised individually for each analyte. Various columns were assessed and sufficient chromatographic separation between the relevant analytes was achieved with an Agilent Technologies Pentafluorophenyl column. Baseline separation was achieved between 25(OH)D3 and 3-epi-25(OH)D3 as well as 25(OH)D2 and 3-epi-25(OH)D2, which is a requirement for this method to be viable. The method was subjected to a series of validation steps to ensure the accuracy and precision of the method. These included the assessment of the analytical range, LOD, LOQ, inaccuracy, imprecision, stability, interference and recovery. It was found that the optimised method had good linearity (r > 0.995), acceptable repeatability (CV < 10 %) and within-lab precision (CV < 15%) and excellent method accuracy (systematic error < 6.60 %). Furthermore, all the analytes proved to be stable for 48 hours after sample preparation with no interferences found for co-eluting analytes. Finally, based on the sigma metric scale specifications, it was calculated that this method proved to be “world class” and very little QC is needed to ensure the quality of the data derived from this method. Based on the findings in this study, it was concluded that a novel LC-MS/MS method for the quantification of twelve vitamin D analogues in a single run was successfully developed. All the LC-MS/MS parameters were optimised to ensure optimal analytical sensitivity for each analyte and the method was validated based on a series of method validation steps required for implementation into a clinical laboratory. This validation proved this method to be ready for implementation into a clinical environment. / MSc (Biochemistry), North-West University, Potchefstroom Campus, 2015

In the early 19th century a ground-breaking discovery was made that linked a dietary deficiency of a fat-soluble vitamin with the childhood disease known as rickets. The vitamin was named vitamin D and extensive research regarding the physiological importance of this vitamin followed ever since. It is currently known that vitamin D plays an important role in maintaining the calcium and phosphate homeostasis in the human body. Less clear evidence states the medical importance of vitamin D in the prevention and cancer, autoimmune disease and diabetes. Current literature shows that vitamin D has five distinct forms, vitamin D1 to D5, of which vitamin D2 and D3 are the most studied forms. The term “vitamin D” is often wrongfully used to include the vitamin D mother molecule, the vitamin D status indicator (25(OH)D), the biologically active form (1,25(OH)2D) and biologically inactive form (24,25(OH)2D). The interest for measurement of these vitamin D analogues is a continuously growing field both on individual and epidemiological level. For decades laboratories have struggled to produce a robust method capable of quantifying these different vitamin D analogues and uncovered a new form of complexity regarding the analysis of these analogues. The identification of the C3-epimeric forms of vitamin D metabolites has forced laboratories to rethink their analytical methods and several concerns were raised regarding the overestimation of the true vitamin D status by current analytical methods. The quantification of the biologically active and inactive forms of vitamin D is reported to be difficult and to date very few LC-MS/MS methods reported in the literature are able to quantify various vitamin D analogues. However, to our knowledge none of these methods are able to include the precursor vitamin D, the 25-hydroxylated metabolites, the biologically active and inactive metabolites, C3-epimers and isobaric compounds in a single run. Therefore the aim of this study was to develop, optimise and validate a LC-MS/MS method for the quantification of twelve vitamin D analogues in a single run. This was done by optimising the underlying LC-MS/MS parameters to ensure optimal analytical sensitivity in positive ESI mode and sufficient chromatographic separation between analytes with similar chemical properties. Furthermore, the optimised method was validated to ensure the accuracy and precision of the method before implementation into a clinical environment. The vitamin D analogues included in this study were vitamin D2, vitamin D3, 25(OH)D2, 25(OH)D3, 1,25(OH)2D2, 1,25(OH)2D3, 24,25(OH)2D2, 24,25(OH)2D3, 3-epi-25(OH)D2, 3-epi- 25(OH)D3, 7(OH)4C3 and 1α(OH)D3. A double liquid-liquid extraction with hexane and ethyl acetate were found to be the most efficient at extracting the vitamin D analogues from a serum matrix after matrix modification with sodium hydroxide. Recoveries of > 95 % (CV <10 %) were achieved for all the analytes. It was noted that a precursor adduct other than the molecular mass ion for a specific vitamin D analogue can produce a more abundant MS1 signal and that the ESI source parameters vary between analytes with different chemical properties and should therefore be optimised individually for each analyte. Various columns were assessed and sufficient chromatographic separation between the relevant analytes was achieved with an Agilent Technologies Pentafluorophenyl column. Baseline separation was achieved between 25(OH)D3 and 3-epi-25(OH)D3 as well as 25(OH)D2 and 3-epi-25(OH)D2, which is a requirement for this method to be viable. The method was subjected to a series of validation steps to ensure the accuracy and precision of the method. These included the assessment of the analytical range, LOD, LOQ, inaccuracy, imprecision, stability, interference and recovery. It was found that the optimised method had good linearity (r > 0.995), acceptable repeatability (CV < 10 %) and within-lab precision (CV < 15%) and excellent method accuracy (systematic error < 6.60 %). Furthermore, all the analytes proved to be stable for 48 hours after sample preparation with no interferences found for co-eluting analytes. Finally, based on the sigma metric scale specifications, it was calculated that this method proved to be “world class” and very little QC is needed to ensure the quality of the data derived from this method. Based on the findings in this study, it was concluded that a novel LC-MS/MS method for the quantification of twelve vitamin D analogues in a single run was successfully developed. All the LC-MS/MS parameters were optimised to ensure optimal analytical sensitivity for each analyte and the method was validated based on a series of method validation steps required for implementation into a clinical laboratory. This validation proved this method to be ready for implementation into a clinical environment. / MSc (Biochemistry), North-West University, Potchefstroom Campus, 2015

Neste estudo foi proposto o desenvolvimento de um software para automatizar o processo de estimativa da incerteza de medição pelo método descrito no Guia EURACHEM. Com a finalidade de testar a eficácia do software, foi desenvolvido um procedimento analítico para a determinação de compostos fenólicos na água subterrânea do Instituto de Pesquisas Energéticas e Nucleares a fim de se obter dados reais de um processo. Para tanto, a determinação dos compostos foi realizada por cromatografia gasosa acoplada ao detector de espectrometria de massas, GC/MS. Para garantir a qualidade dos dados gerados, o procedimento analítico foi submetido ao processo de validação, onde foram avaliados os parâmetros: seletividade/especificidade, faixa de trabalho e faixa linear de trabalho, linearidade, limite de detecção, limite de quantificação, precisão, exatidão, recuperação e robustez. A estimativa da incerteza da medição foi realizada pelo software desenvolvido e manualmente, confirmando que o mesmo é adequado para o processo. Além disso, o software foi testado utilizando dados da literatura, o que confirmou sua eficácia. Os resultados da análise da água subterrânea demonstraram que não há a presença de compostos fenólicos nos níveis estudados. A utilização de sistemas automatizados para a estimativa da incerteza diminui ou minimiza erros sistemáticos e permite trabalhar com mais organização e controle do processo. / A software development to automate the process of uncertainty measurement by the method described by the Guide EURACHEM is proposed. In order to test the effectiveness of the software, an analytical procedure for phenolic compounds determination in the ground water was developed. Water samples were collected at Instituto de Pesquisas Energéticas e Nucleares (IPEN, São Paulo, Brazil) campus area. The determination of compounds was performed by gas chromatography coupled to mass spectrometry detector (GC / MS). To ensure the quality of the generated data, the analytical procedure was submitted to a validation process. The evaluated parameters were: selectivity / specificity, working range and linear range, linearity, detection limit, quantification limit, precision, recovery and robustness. The uncertainty measurements were performed by the software and manually, confirming that it is suitable for the process. Moreover, the software was tested using literature data, which confirmed its effectiveness. The results of the ground water analysis showed that there are no phenolic compounds within the studied levels. The use of automated systems for the uncertainty estimation is very promising because it reduces or minimizes systematic errors and allows working in a more organized way and with the process under control.

Neste estudo foi proposto o desenvolvimento de um software para automatizar o processo de estimativa da incerteza de medição pelo método descrito no Guia EURACHEM. Com a finalidade de testar a eficácia do software, foi desenvolvido um procedimento analítico para a determinação de compostos fenólicos na água subterrânea do Instituto de Pesquisas Energéticas e Nucleares a fim de se obter dados reais de um processo. Para tanto, a determinação dos compostos foi realizada por cromatografia gasosa acoplada ao detector de espectrometria de massas, GC/MS. Para garantir a qualidade dos dados gerados, o procedimento analítico foi submetido ao processo de validação, onde foram avaliados os parâmetros: seletividade/especificidade, faixa de trabalho e faixa linear de trabalho, linearidade, limite de detecção, limite de quantificação, precisão, exatidão, recuperação e robustez. A estimativa da incerteza da medição foi realizada pelo software desenvolvido e manualmente, confirmando que o mesmo é adequado para o processo. Além disso, o software foi testado utilizando dados da literatura, o que confirmou sua eficácia. Os resultados da análise da água subterrânea demonstraram que não há a presença de compostos fenólicos nos níveis estudados. A utilização de sistemas automatizados para a estimativa da incerteza diminui ou minimiza erros sistemáticos e permite trabalhar com mais organização e controle do processo. / A software development to automate the process of uncertainty measurement by the method described by the Guide EURACHEM is proposed. In order to test the effectiveness of the software, an analytical procedure for phenolic compounds determination in the ground water was developed. Water samples were collected at Instituto de Pesquisas Energéticas e Nucleares (IPEN, São Paulo, Brazil) campus area. The determination of compounds was performed by gas chromatography coupled to mass spectrometry detector (GC / MS). To ensure the quality of the generated data, the analytical procedure was submitted to a validation process. The evaluated parameters were: selectivity / specificity, working range and linear range, linearity, detection limit, quantification limit, precision, recovery and robustness. The uncertainty measurements were performed by the software and manually, confirming that it is suitable for the process. Moreover, the software was tested using literature data, which confirmed its effectiveness. The results of the ground water analysis showed that there are no phenolic compounds within the studied levels. The use of automated systems for the uncertainty estimation is very promising because it reduces or minimizes systematic errors and allows working in a more organized way and with the process under control.

Submitted by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-09-22T13:21:31Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) BARBRA KATYÚSCYA SANCHES MORAES.pdf: 1057225 bytes, checksum: 43106c269cbed3f116c11fc89abec2f5 (MD5) / Approved for entry into archive by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-09-22T13:22:21Z (GMT) No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) BARBRA KATYÚSCYA SANCHES MORAES.pdf: 1057225 bytes, checksum: 43106c269cbed3f116c11fc89abec2f5 (MD5) / Made available in DSpace on 2016-09-22T13:22:21Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) BARBRA KATYÚSCYA SANCHES MORAES.pdf: 1057225 bytes, checksum: 43106c269cbed3f116c11fc89abec2f5 (MD5) Previous issue date: 2015-05-29 / Neste estudo, um método de CLAE-DAD foi desenvolvido e validado para a determinação de LASSBio-1736 em plasma de rato usando diclofenaco de sódio como padrão interno (PI). Extração líquido-líquido com acetonitrila foi utilizado para extrair LASSBio-1736. A separação cromatográfica foi realizada com coluna Waters Spherisorb®S5 ODS2 C18 (150 mm x 4,6 mm, 5μm), fase móvel isocrática composta por Trietilamina 0,3% (pH 4), metanol e acetonitrila (45:15:40, v/v/v) com vazão de 1 mL/min. Ambos LASSBio-1736 e PI foram eluídos em 4,2 e 5 min, respectivamente. O limite inferior de quantificação foi de 0,2 μg/mL e linearidade entre 0,2 - 4 μg/mL, com um r2> 0,99. A exatidão do método foi > 90,5%. Os desvios padrão relativos intra e inter-dias foram < 6,19 e < 7,83%, respectivamente. O método mostrou sensibilidade, linearidade, precisão, exatidão e seletividade necessária para quantificar LASSBio-1736 em estudos farmacocinéticos pré-clínicos O presente trabalho também investigou a farmacocinética plasmática e a distribuição do LASSBio-1736 em ratos Wistar. A farmacocinética de LASSBio-1736 foi investigada após a administração de dose intravenosa (3,2 mg/kg), por via oral e intraperitoneal (12,6 mg/kg).. A distribuição nos tecidos foi avaliada após administração de dose i.v. bolus. Os resultados para a via intravenosa indicam longo tempo meia-vida (24,3 ± 8,2 h), depuração de 49,3 ± 9,8 mL/Kg*h e volume de distribuição de 1,16 ± 0,3 L/kg. Para a via oral o tempo meia-vida foi de 28,6 ± 4,6 h, depuração de 49,7 ± 13 mL/Kg*h e volume de distribuição de 1,47 ± 0,34 L/kgforam semelhantes e biodisponibilidade de 12%. Para a via intraperitoneal o tempo meia-vida foi de 26 ± 8,9 h, depuração de 58 ± 19,6 mL/Kg*h e volume de distribuição de 1,8 ± 0,8 L/kg e biodisponibilidade de 38%. O LASSBio-1736 demonstrou penetração tecidual adequada no fígado, baço e pele. Com base nas características farmacocinéticas de outros fármacos leishmanicidas e a proposição de decisão para estudos farmacocinéticos visando a descoberta de candidatos a fármacos e a continuidade dos estudos, o LASSBio-1736 possui características farmacocinéticas apropriadas para um medicamento leishmanicida e novos estudos devem ser realizados para o escalonamento interespécies, além de estudos farmacológicos e toxicológicos complementares. / In this study, a method was developed and validated for HPLC-PDA determination LASSBio-1736 in rat plasma using diclofenac sodium as internal standard (IS). Liquid-liquid extraction was used to extract acetonitrile LASSBio-1736. The chromatographic separation was performed with Waters Spherisorb®S5 ODS2 C18 column (150 mm x 4.6 mm, 5μm), isocratic mobile phase consisting of Triethylamine 0.3% (pH 4), methanol and acetonitrile (45:15:40, v / v / v) with a flow rate of 1 mL/min. Both LASSBio-1736 and IS were eluted at 4.2 and 5 min, respectively. The lower limit of quantification was 0.2 μg/mL and linearity between 0.2 - 4 μg/mL, with r2> 0.99. The accuracy was > 90.5%. The relative standard deviation within and between days were < 6.19 and < 7.83%, respectively. The method showed sensitivity, linearity, precision, accuracy and selectivity needed to quantify LASSBio-1736 in preclinical pharmacokinetic studies. This study also investigated the plasma pharmacokinetics and distribution of LASSBio-1736 in Wistar rats. The pharmacokinetic LASSBio-1736 was investigated after intravenous dose administration (3.2 mg/kg) intraperitoneally and orally (12.6 mg/kg). The tissue distribution was evaluated after iv bolus dose administration. The results indicated an intravenous long half-life (24.3 ± 8.2 h) clearance 49.3 ± 9.8 mL/kg*h and the volume of distribution 1.16 ± 0.3 L/ kg. The oral route the half-life was 28.6 ± 4.6 h, clearance 49.7 ± 13 mL/kg*h and volume of distribution 1.47 ± 0.34 L/kg and bioavailability of 12%. The intraperitoneally half-life was 26 ± 8.9 h, clearance 58 ± 19.6 mL/kg*h and the volume of distribution 1.8 ± 0.8 L/kg and bioavailability of 38%were similar. The LASSBio-1736 showed adequate tissue penetration for liver, spleen and skin. Based on the pharmacokinetic characteristics of other antileishmanial drugs and the decision proposition for pharmacokinetic studies for the discovery of drug candidates and continuing studies, the LASSBio-1736 has pharmacokinetic characteristics appropriate for a leishmanicide and new drug studies should be conducted to the interspecies scaling, and additional pharmacological and toxicological studies.

A gentamicina é um complexo antibiótico de largo espectro, produzido por actinomicetos do gênero Micromonospora e classificado entre os antibióticos aminoglicosídeos, utilizado no tratamento de infecções graves, devidas a microrganismos Gram-negativos. Alterações da sua atividade antimicrobiana, não demonstradas pelos ensaios químicos, podem ser avaliadas pelos ensaios microbiológicos. O objetivo deste trabalho foi comparar os delineamentos experimentais 5 x 1, 2 x 2 e 3 x 1, avaliando-se os parâmetros de validação de especificidade, linearidade, faixa ou intervalo, precisão e exatidão para cada delineamento experimental em diferentes níveis de concentração, apresentações e lotes. O plano de trabalho constituiu-se na realização de 81 ensaios (em 3 réplicas) de doseamento microbiológico de gentamicina. As concentrações das soluções empregadas foram preparadas numa faixa de 1,0 µg/mL a 5,0 µg/mL, diluídos em tampão fosfato 0,1 M pH 8,0. O meio utilizado foi o meio antibiótico no. 11, com Staphyloccocus epidermidis (ATCC 12228). Empregou-se 21 mL de meio como camada base e 4 mL de meio inoculado à 1% como camada superfície. As placas foram incubadas por 16-18 horas à 37 ± 1 °C. Os três delineamentos empregados apresentaram especificidade adequada para análise de creme dermatológico e solução injetável contendo sulfato de gentamicina. Também apresentaram exatidão e linearidade no intervalo avaliado. Os delineamentos não apresentaram diferença significativa quanto a precisão. Os resultados foram comparados através da determinação de índices de capacidade do sistema de medição. A análise estatística demonstrou que não há diferença significativa entre os resultados obtidos pelos delineamentos 5 x 1, 2 x 2 e 3 x 1, sendo equivalentes e intercambiáveis. / Gentamicin is a broad-spectrum antibiotic complex produced by actinomycetes belonging to Micromonospora genus and classified among aminoglycoside antibiotics, used in the treatment of serious infections derived from Gram-negative microorganisms. Alterations of their antimicrobial activity not shown in chemical assays can be evaluated through microbiological assays. The aim of this work was to compare 5 x 1, 2 x 2 and 3 x 1 experimental designs, evaluating validation parameters of specificity, linearity, range, precision, and accuracy for each experimental design in different levels of concentration, presentation, and lots. It consisted of 81 assays (in 3 replicas) of gentamicin microbiological dosage. The concentrations of the solutions used were employed in a range from 1.0 µg/ml to 5.0 µg/ml, diluted in phosphate buffer 0.1 M pH 8.0. Antibiotic medium number 11 was used, with Staphyloccocus epidermis (ATCC 12228)21ml of medium were used as base layer and 4 ml of medium inoculated at 1% were used as surface layer. The plates were incubated for 16-18 hours at 37 ± 1 ºC. The three designs employed showed adequate specificity for analysis of dermatological cream and injectable solution containing gentamicin sulphate. They also showed accuracy and linearity in the range evaluated, but not a significant difference concerning precision. The results were compared by means of the determination of the rates of measurement system capacity. The statistical analysis demonstrated that there is no significant difference among the results obtained.