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Chromatographic and Spectroscopic Studies on Aquatic Fulvic AcidChang, David Juan-Yuan 08 1900 (has links)
High Performance Liquid Chromatography (HPLC) was used to investigate the utility of this technique for the analytical and preparative separation of components of aquatic fulvic acids (FA). Three modes of HPLC namely adsorption, anion exchange and reversed phase were evaluated. Aquatic fulvic acids were either extracted from surface water and sediment samples collected from the Southwest of the U.S., or were provided in a high purity form from the USGS. On the adsorption mode, a major fraction of aquatic fulvic acid was isolated on a semipreparative scale and subjected to Carbon-13 NMR and FAB Mass Spectroscopy. Results indicated that (1) The analyzed fraction of fulvic acid contains more aliphatic than aromatic moieties; (2) Methoxy, carboxylic acids, and esters are well-defined moieties of the macromolecule; (3) Phenolic components of the macromolecules were not detected in the Carbon-13 NMR spectrum possibly because of the presence of stable free radicals. Results of the anion exchange mode have shown that at least three types of acidic functionalities in aquatic fulvic acid can be separated. Results also indicated that aquatic fulvic acid can be progressively fractionated by using subsequent modes of HPLC. Results of reversed phase mode have shown that (1) The fractionation of aquatic fulvic acid by RP-HPLC is essentially controlled by the polarity and/or pH of the carrier solvent system; (2) Under different RP-HPLC conditions aquatic fulvic acid from several locations are fractionated into the same major components; (3) Fulvic acid extracted from water and sediment from the same site are more similar than those extracted from different sites; (4) Cationic and anionic ion pair reagents indicated the presence of amphoteric compounds within the polymeric structure of fulvic acid. Each mode of HPLC provided a characteristic profile of fulvic acid. The results of this research provided basic information on the behavior of aquatic fulvic acids under three modes of HPLC. Such informations are prerequisite for further investigation by spectroscopic methods.
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Rapid Isolation and Purification of Plasmid DNA Using High Performance Liquid ChromatographyNam, Kiebang 05 1900 (has links)
High Performance Liquid Chromatography (HPLC) has been employed as an analytical tool for the purification and separation of nucleic acids. A Nucleogen DEAE 4000-10 weak anion exchange column, prepacked with modified silica gels, was used to purify and separate a number of Escherichia coli plasmids. Plasmid DNAs were extracted by the alkaline lysis method. The cleared lysate was injected directly onto the Nucleogen column, and the peaks were collected, desalted and analysed by gel electrophoresis. On the chromatogram, the pBR322 formed a distinctive peak at 27 minutes and partial separation was made for the E. coli V517 plasmids. Plasmid pBR322 showed a clear band without any detectable contamination on agarose gel. This purified plasmid DNA is biologically active for enzymatic reaction commonly used in genetic engineering techniques.
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High performance liquid chromatographic determination of (-)-epicathechin in cocoa beans and the effects of varietal types, curing, and roasting on its concentrationKim, Henry. January 1982 (has links) (PDF)
Thesis (Ph. D.)--Pennsylvania State University, 1982. / Includes bibliographical references.
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TOWARDS BIOMARKER DISCOVERY IN CONGENITAL URINARY TRACT OBSTRUCTIONOrton, Dennis 09 May 2014 (has links)
Proteome analysis techonologies are commonly employed for discovery-based biomarker identification studies. This thesis aims to help bridge the gap between analytical technology development and clinical application by improving and appling a proteomics workflow for biomarker discovery in congenital urinary tract obstruction (UTO). By accentuating the importance of experimental design, and evaluating the biological relevance of quantitative proteome analyses, the results of this research provide confidence in a number of identified candidate biomarkers of UTO.
A sensitive method for quantification of proteome samples was developed using temperature controlled reversed-phase liquid chromatography (TPLC). The TPLC system provides high recovery (> 90 %), as well as high accuracy and precision in estimating the concentration across a number of protein sample types (CV < 10 %).
The need for extensive fractionation strategies coupled with LC-MS analysis challenges the throughput of the overall experiment. Development of a dual column LC-MS interface reduced the total analysis time by a factor of 2 over conventional single column LC-MS systems. The system was applied to a quantitative proteome analysis of proximal tubule cells exposed to mechanical stretch, mimicking the conditions they experience during UTO and a urinary exosomal proteome analysis for candidate biomarker identification of this disease.
A total of 1636 proteins were identified in the whole cell proteome analysis, of which 317 were found to be significantly altered in abundance. Analysis of the urinary exosomal proteome yielded 318 proteins, of which 189 were found to be altered in abundance due to obstruction. Western blot confirmation of a few select proteins provided backing to the quantitative proteome analysis, while gene ontology and KEGG pathway analysis yielded functional information.
The results from the quantitative analyses of the urinary exosomes and proximal tubule cells identified candidates for both diagnosis and prognosis of UTO. In addition, activation of a novel pathway was identified, presenting a potential drug target which could be exploited to improve recovery of children following relief of UTO. This thesis therefore contributes useful technological and methodological advancements towards routine proteome analysis, as well as providing candidate biomarker identification for the leading cause of renal functional loss in children.
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High-performance liquid chromatography analysis of fatty acids and mathematical modeling of liquid chromatographyLi, Zhiguo. January 2001 (has links)
Thesis (Ph. D.)--Ohio University, March, 2001. / Title from PDF t.p.
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Hyphenated HPLC-MS technique for analysis of compositional monosaccharides of transgenic corn glycoprotein and characterization of degradation products of diazinon, fonofos and aldicarb in various oxidation systemsWang, Tongwen, January 2007 (has links) (PDF)
Thesis (Ph. D.)--University of Missouri--Rolla, 2007. / Vita. The entire thesis text is included in file. Title from title screen of thesis/dissertation PDF file (viewed April 23, 2008) Includes bibliographical references.
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Separation and identification of peptides by integrated multidimensional liquid chromatography-mass spectrometry (IMDLC-MS)Adusumilli, Harika. January 2007 (has links)
Thesis (M.S.)--University of Missouri-Columbia, 2007. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on April 15, 2008) Vita. Includes bibliographical references.
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Design and development of acquisition, control and processing software for two dimensional high performance liquid chromatographyToups, Erich P. January 2004 (has links)
Thesis (MSc. (Hons.))--University of Western Sydney, 2004. / A thesis presented to the University of Western Sydney, College of Science, Technology and Environment, School of Science, Food and Horticulture, in fulfilment of the requirements for the degree of Master of Science (Honours). Includes bibliographies.
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The synthesis and characterization of reversed phase stationary phases for high performance liquid chromatographyBarnes, Karen Wink. January 1986 (has links)
Thesis (Ph. D.)--University of Florida, 1986. / Description based on print version record. Typescript. Vita. Includes bibliographical references (leaves 156-161).
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Desenvolvimento e validação de método analítico empregando DLLME e HPLC/UV para determinação de benzodiazepínicos em amostra de água / Development and validation of analytical method employing DLLME and HPLC/UV for the determination of benzodiazepines in water sampleMarques, Tamires Valim, 1987- 02 December 2014 (has links)
Orientador: Rodnei Bertazzoli / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Mecânica / Made available in DSpace on 2018-08-24T13:08:38Z (GMT). No. of bitstreams: 1
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Previous issue date: 2014 / Resumo: A crescente preocupação com a poluição das águas por novos poluentes denominados emergentes tem se intensificado, visto que aumentou o número destes compostos detectados em água. Dentre estes compostos encontram-se os fármacos e produtos de higiene pessoal, usados cotidianamente pela população. O presente trabalho teve como objetivo o desenvolvimento de um método simples, rápido e sensível utilizando a microextração líquido líquido (DLLME) combinada com a cromatografia líquida de ultra eficiência (HPLC) com detecção ultravioleta (UV) para a determinação de alguns benzodiazepínicos (bromazepam, clonazepam e diazepam) em amostras de água. A determinação foi realizada em uma coluna C18 de acordo com as condições cromatográficas ótimas (fase móvel acetonitrila:água (60:40, v/v); vazão 1,2 mL min-1; detecção 225 nm). No método de extração, uma mistura apropriada de solvente extrator e dispersor foi injetada rapidamente na amostra aquosa (10 mL) com auxílio de uma seringa. De modo que uma solução turva foi formada, esta solução é caracterizada por conter pequenas partículas do solvente extrator que se dispersa na fase aquosa. Os parâmetros da extração, tais como natureza e volume dos solventes extrator e dispersor, tempo de extração, pH da amostra, força iônica, velocidade e tempo de centrifugação, foram estudados para a otimização. Com as condições ótimas definidas (solvente extrator: clorofórmio, 200 ?L; solvente dispersor: metanol, 700 ?L; pH da amostra 9,0; velocidade e tempo de extração: 5000 rpm, 1 minuto; força iônica: adição de 1% (p/v) de (NH4)2SO4) o método proposto foi validado seguindo as figuras de método preconizadas pela ANVISA na Resolução N° 899 de 2003. A faixa linear para cada fármaco foram 8,0 - 96 ?g L-1 para bromazepam, 4,0 - 48 ?g L-1 para clonazepam e 1,0 - 12 ?g L-1 para diazepam. Todas as curvas obtiveram valores de (r) superiores a 0,999. Os limites de detecção e quantificação obtidos foram 2,4 e 8,0 ?g L-1 para bromazepam, 1,2 e 4,0 ?g L-1 para clonazepam, 0,2 e 1,0 ?g L-1 para diazepam, respectivamente. As recuperações variaram de 50 a 110% com RSD (Desvio Padrão Relativo) inferiores a 12,7 %. Finalmente, o método proposto foi aplicado em amostras coletadas na represa Billings localizada no município de Diadema-SP. / Abstract: The growing concern over water pollution caused by so-called new emerging pollutants has intensified since increased the number of these compounds detected in water. Among these compounds are pharmaceuticals and personal care products used daily by the population. This study aimed to develop a simple, rapid and sensitive liquid using liquid micro extraction (DLLME) combined with ultra-performance liquid chromatography (HPLC ) with ultraviolet detection (UV) for the determination of some benzodiazepines (bromazepam, clonazepam and diazepam) in water samples . The determination was performed on a C18 column in accordance with the optimal chromatographic conditions (mobile phase acetonitrile: water (60:40, v/v), flow rate 1.2 mL min-1, detection 225 nm). In the extraction method, a suitable mixture of extractant and dispersing solvent was injected rapidly into the aqueous sample (10 ml) with a syringe. So that a cloudy solution was formed, this solution is characterized by containing fine drops of the extractor solvent is dispersed in the aqueous phase. The parameters of the extraction, such as the nature and volume of the extractor and disperser solvents, extraction time, sample pH, ionic strength, speed and time of centrifugation, were studied for optimization. With the defined optimal conditions (extracting solvent: chloroform, 200 ?L; disperser solvent: methanol, 700 ?L, sample pH 9.0, extraction time and speed: 5000 rpm, 1 minute; ionic strength: adding 1% (p/v) (NH4)2SO4) the proposed method was validated following the figures of merit recommended by the ANVISA Resolution No. 899 of 2003. The linear ranges for each drug were 8.0 to 96 ?g L- 1 for bromazepam, 4.0 to 48 ?g L- 1 for clonazepam and 1.0 to 12 ?g L- 1 for diazepam. All curves obtained values (r) greater than 0.999. The limits of detection and quantification obtained were 2.4 and 8.0 ?g L- 1 to bromazepam, 1.2 and 4.0 ?g L- 1 to clonazepam, 0.2 and 1.0 ?g L- 1 for diazepam, respectively. Recoveries ranged from 50 to 110% with RSD (Relative Standard Deviation) of less than 12.7%. Finally, the proposed method was applied to samples collected in the Billings dam located in Diadema-SP. / Mestrado / Materiais e Processos de Fabricação / Mestre em Engenharia Mecânica
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