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Analysis of flavan-3-ols by capillary electrophoresis.January 2004 (has links)
Lee Wai Hang. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 71-76). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.ii / Table of Contents --- p.v / Abbreviations --- p.viii / List of Figures --- p.ix / List of Tables --- p.xiv / Chapter / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- The French Paradox --- p.1 / Chapter 1.2 --- Flavonoids --- p.2 / Chapter 1.3 --- Grape seed extract --- p.4 / Chapter 1.4 --- Instrunmental analysis --- p.7 / Chapter 1.4.1 --- High Performance Liquid Chromatography --- p.7 / Chapter 1.4.2 --- Colorimetry --- p.9 / Chapter 1.5 --- Capillary Electrophoresis --- p.10 / Chapter 1.5.1 --- Instrunmentation --- p.10 / Chapter 1.5.2 --- Electroosmotic Flow --- p.11 / Chapter 1.5.3 --- Electrophoretic mobility --- p.13 / Chapter 1.6 --- Objective of the study --- p.15 / Chapter 2. --- Experimental --- p.18 / Chapter 2.1 --- Reagents and material --- p.18 / Chapter 2.1.1 --- Reagents --- p.18 / Chapter 2.1.2 --- Instrunmentation --- p.18 / Chapter 2.1.3 --- Reference compounds --- p.19 / Chapter 2.1.4 --- Samples --- p.19 / Chapter 2.2 --- Selection of solvent for sample preparation --- p.20 / Chapter 2.3 --- Procedures --- p.21 / Chapter 2.3.1 --- Preparation of running buffer solution --- p.21 / Chapter 2.3.2 --- Preparation of standard solution --- p.21 / Chapter 2.3.3 --- Preparation of sample solution --- p.22 / Chapter 2.3.4 --- Flushing procedures --- p.22 / Chapter 3. --- Results and Discussion --- p.24 / Chapter 3.1 --- Preliminary experiments --- p.24 / Chapter 3.2 --- Effect of pH --- p.27 / Chapter 3.3 --- Addition of surfactant --- p.30 / Chapter 3.4 --- Effect of SDS concentration --- p.35 / Chapter 3.5 --- Addition of cyclodextrins --- p.39 / Chapter 3.6 --- Urea --- p.46 / Chapter 3.7 --- Addition of organic modifier --- p.47 / Chapter 3.8 --- Effect of borate concentration --- p.49 / Chapter 3.9 --- Effect of cyclodextrin concentration --- p.53 / Chapter 3.10 --- Optimized condition --- p.58 / Chapter 3.11 --- Reproducibility of the method --- p.58 / Chapter 3.12 --- Quantitative analysis of reference compounds --- p.60 / Chapter 3.13 --- Application of the CE method in grape seed products --- p.62 / Chapter 4. --- Conclusion --- p.69 / References --- p.71
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Preformulation and mechanistic studies on inclusion complexes of selected flavonoids with beta-cyclodextrin and its water-soluble derivatives. / Preformulation and mechanistic studies on inclusion complexes of selected flavonoids with b-cyclodextrin and its water-soluble derivatives / CUHK electronic theses & dissertations collection / Digital dissertation consortiumJanuary 2005 (has links)
"December 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 221-233) / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.
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The identification of the precursor of off odor causing compounds, produced during storage of ground pearl millet (Pennisetum americanum (L) Leeke)Reddy, Vijaya P. January 1985 (has links)
Call number: LD2668 .T4 1985 R422 / Master of Science
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Bio-active compounds isolated from mistletoe (Scurulla oortiana (Korth.) Danser) parasitizing tea plant (Camellia sinensis L.)Kirana, Chandra. January 1996 (has links) (PDF)
Bibliography: leaves 87-96. This thesis investigates non-proteinaceous low molecular weight flavonoid and alkaloid compounds in Scurulla oortiana (Korth.) Danser grown on Camellia sinens. Three flavonols are identified in S. oortiana (Korth.) Danser growing on different hosts. The identification and characterisation of these flavonoids are carried out using various chromatographic and spectrometric procedures. Two purine alkaloids are isolated from and identified in S. oortiana (Korth.) Danser parasitizing tea plant, C. Sinensis. The antifungal activity of the phenolic compounds isolated from mistletoe parasitizing tea plant is examined.
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Separation and characterization of glycosylated phenolic compounds and flavonoids from maple productsCôté, Jacinthe January 2003 (has links)
Using a model system of glycosylated and aglycon standards consisting of rutin and quercetin respectively, and a series of pre-packed solid phase extraction cartridges, including C18 Extra-Clean, DSC-18, DPA-6S, Oasis HLB and Amberlite XAD-2. The experimental findings also showed that use of a commercial hesperinidase preparation, resulted in adequate hydrolysis of the glycosylated standard rutin. Based on these findings, the phenolic compounds and flavonoids from maple sap and syrup were separated using the Amberlite XAD-2 column, where the glycosylated fractions eluted with 60% aqueous methanol solution and the aglycon fractions eluted with a methanol:acetonitrile mixture (1:1, v/v). The recovered glycosylated fractions were subjected to enzymatic hydrolysis using the hesperinidase preparation and the liberated phenolic compounds and flavonoids, as well as the sugar components were analyzed by high-performance liquid chromatography (HPLC).
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Recovery, separation and characterization of phenolic compounds and flavonoids from maple productsDeslauriers, Isabelle. January 2000 (has links)
Comparative high-performance liquid chromatography (HPLC) and gas-liquid chromatography (GC) analyses of selected phenolic and flavonoid standards were developed using a wide range of detectors, including ultraviolet diode-array (UV-DAD) and electrochemical (EC) detectors for HPLC and flame ionization detector (FID) and mass spectrometry (MS) for GC. The results demonstrated that the limits of detection obtained with HPLC-EC analysis were 10 to 500-times higher for phenolic acid standards and 2 to 50-times higher for flavonoid standards than those obtained with the HPLC-UV analysis. HPLC-EC was more sensitive than GC/FID for all investigated standards, especially for vanillin and syringaldehyde. The results indicated that GC/FID/MS analysis of phenolic and flavonoid standards was more efficient than that of HPLC, providing a fast analysis with better resolution and baseline separation of all standards with minimum co-elution. The only co-elution encountered in GC/FID was with coniferol and p-coumaric acids. For HPLC analysis, (-)-epicatechin, caffeic and homovanillic acids were co-eluted at 28.04 min and sinapic and ferulic acids at 34.57 min. Phenolic compounds and flavonoids were extracted from maple sap and maple syrup with ethyl acetate and the recovered compounds were subjected to HPLC and GC analyses. Tentative identification of phenolic compounds and flavonoids in maple sap and maple syrup indicated the presence of protocatechuic acid, hydroxycinnamic acid derivatives, (+)-catechin, (-)-epicatechin, vanillin, coniferol, syringaldehyde, flavanols and dihydroflavonols related compounds. In addition, the identification by GC/MS of protocatechuic acid, vanillin, syringaldehyde, coniferol and p-coumaric acid was made by comparing mass spectrum characteristics of individual peak from total ion chromatogram (TIC) to that of standard compounds. The seasonal variation of selected phenolic compounds and flavonoids present in maple sap and maple syrup was also invest
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Pharmaceutical analysis and aspects of the quality control of St. John's Wort productsWild, Tracy Joy January 2003 (has links)
Most complementary medicines contain a multitude of chemical components, some of which are claimed to contribute to the biological activity of such products. Use of complementary medicines for preventative and therapeutic purposes is increasing rapidly worldwide. Unfortunately, although control of these products is essential to ensure quality, safety, and efficacy, the quality control of most herbal preparations is currently poor to non-existent, with little or no safety and efficacy data required to support the marketing and use of these products. The objective of this study was therefore to develop suitable analytical methods to qualitatively and quantitatively analyse the relevant components (rutin, isoquercitrin, hyperoside, quercitrin, quercetin, kaempferol, hypericin, pseudohypericin and hyperforin) in St John's Wort dosage forms for quality control purposes. A gradient HPLC method using a Luna 5·mC₁₈(2) 150 x 2.00mm internal diameter (i.d.) column and UV detection, was developed for the separation of six of the relevant flavonoid compounds in St John's Wort, namely rutin, isoquercitrin, hyperoside, quercitrin, quercetin and kaempferol. The development process involved a systematic investigation of gradient conditions, flow rate, and temperature. This method was subsequently applied to assay selected commercially available St John's Wort products. This system provided the necessary accuracy, precision and reproducibility and was associated with several advantages when compared to using standard bore (4.60 mm i.d.) HPLC columns. The method developed is currently the only known method that separates all six relevant flavonoids in a reasonable run time (less than 20 minutes). It is also one of the few methods that has sufficient separation between rutin, isoquercitrin and hyperoside. A qualitative method for the fingerprinting of flavonoid components was also developed, using capillary electrophoresis (CE). CE is a rapidly growing powerful analytical technique for the separation of charged compounds. Micellar electrokinetic chromatography (MEKC) is a very powerful electrophoretic technique that is capable of selectively resolving both neutral and ionic solutes in a single run. A MEKC method suitable for the separation and determination of various flavonoid constituents used as marker compounds in Hypericum perforatum was developed. Investigations into the effect of pH, ionic strength, applied voltage and capillary dimensions on separation were performed. The optimised method was then applied to qualitatively analyse various St John's Wort products on the market. This method was found to be advantageous in that it was simple, cost-effective, required minimal sample preparation and utilised very small quantities of sample. Due to the vast differences in chemical properties between the various marker and active components in St John's Wort, it was necessary to develop separate analytical methods for the flavonoids and for the other three relevant compounds (hypericin, pseudohypericin and hyperforin). An isocratic HPLC method using a Luna 5·mC₁₈(2) 150 x 2.00mm (i.d.) column and UV detection was developed for the separation of hypericin, pseudohypericin and hyperforin. The development process involved a systematic investigation of buffer molarity, mobile phase composition, pH, flow rate, and temperature. This method was subsequently applied to assay selected commercially available St John's Wort products on the South African market. This system also provided the necessary accuracy, precision and reproducibility, as well as the advantages associated with the use of a narrow bore column as opposed to the use of the more commonly used wider bore columns. This method was validated and used to quantitate these three compounds in various commercial St John's Wort products. By applying this method to liquid chromatography – tandem mass spectrometry (LC-MS-MS), qualitative analyses of the same products was performed to obtain confirmation of the quantitative HPLC results. Mass spectrometry is a powerful detection tool that is more selective and specific than many detection systems used with HPLC. Natural medicines usually constitute a multitude of constituents with much potential interference. In this regard LC-MS-MS is a powerful tool, with its ability to unequivocally identify target analytes regardless of the presence of interferences or complex matrices. ESI-MS-MS was used for the qualitative analysis of the content of the naphthodianthrones and hyperforin in the respective tablet products assayed with HPLC. LC-MS-MS analyses were performed in order to identify the constituents and to verify the specificity of the HPLC method. High inter-product and inter-batch variability was observed for all nine compounds assayed. These quantitative results were confirmed with the respective qualitative analyses. This study confirms the need for strict quality control of herbal medicinal products commercially available to consumers.
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Recovery, separation and characterization of phenolic compounds and flavonoids from maple productsDeslauriers, Isabelle. January 2000 (has links)
No description available.
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Separation and characterization of glycosylated phenolic compounds and flavonoids from maple productsCôté, Jacinthe January 2003 (has links)
No description available.
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Análise de flavanóides por cromatografia líquida de alta eficiência e eletroforese capilar - otimização de separação e aplicações tecnológicas / Flavonoids Analysis by high-performance liquid chromatography and capillary electrophoresis - separation optimization and technological applicationsTonin, Fernando Gustavo 09 June 2006 (has links)
No presente trabalho foram estudadas as separações de 18 flavonóides (9 agliconas e 9 glicosídeos) pelas técnicas de Cromatografia Líquida de Alta Eficiência em fase reversa (RP-HPLC) e Cromatografia Micelar Eletrocinética em fluxo reverso (RF-Meck). Em ambas as técnicas foram avaliados solventes puros (metanol, acetonitrila e tetrahidrofurano) e suas misturas como formas de promover a variação de seletividade, através da modificação da fase móvel em HPLC, e da natureza do aditivo orgânico em RF-Meck. Nos estudos efetuados em HPLC utilizando-se gradiente, pode-se comprovar a possibilidade da modelagem do fator de retenção em funçã da proporção de solvente utilizados (MeOH, ACN, THF e suas misturas). Pode-se ainda, com base nos dados de retenção e na análise hierárquica de c1usters, diferenciar quatro diferentes grupos de sistemas cromatográficos com diferentes seletividades para flavonóides agliconas, e outros quatro com diferentes seletividades para glicosídeos. Os sistemas cromatográficos mais ortogonais (cada um pertencente a um grupo de seletividade) foram aplicados na separação de uma planta modelo (Azadirachta indica), de onde pode-se escolher a fase móvel mais seletiva para se otimizar a separação dos flavonóides glicosilados presentes nas folhas desta planta. No método final otimizado pode-se identificar e quantificar cinco dos flavonóides majoritários presentes, sendo três glicosídeos de quercetina (rutina, isoquercitrina e quercitrina) e dois glicosídeos de kaempferol (astragalin e nicotiflorin), em amostras de duas diferentes procedências (Piracicaba-SP e Silvânia-GO). Nos estudos envolvendo a separação dos dezoito flavonóides por RFMEKC pode-se comprovar diferenças significativas de seletividade quando se varia a natureza do solvente orgânico utilizado como aditivo, além de se observar tendências na migração em função das propriedades do solvente adicionado e da estrutura molecular do flavonóide. O solvente de menor eficiência para separação dos flavonóides foi o MeOH. Através da análise dos eletroferogramas obtidos através de um planejamento experimental de misturas, e das trocas de pares críticos observadas nos vários eletrólitos utilizados, obteve-se um método de separação com apenas um par crítico em menos de 12 minutos de corrida. O coeficiente de variação obtido para o fator de retenção foi de 1,5% e para área de 3%, considerando-se cinco injeções. O método desenvolvido foi aplicado com sucesso na identificação dos flavonóides majoritários presentes na planta modelo (Neem), obtendo-se o mesmo resultado do estudo anterior. Como forma de avaliar a concentração de flavonóides totais presentes em espécies vegetais é comum a análise de extratos após hidrólise ácida (conversão de todos glicosídeos em agliconas). Desta forma otimizou-se uma metodologia de separação em RP-HPLC de 8 flavonóides agliconas comumente presentes em alimentos e extratos vegetais de uso cosmético. A otimização foi efetuada mediante um planejamento experimental de misturas, para escolha da fase móvel mais seletiva, e de um planejamento fatorial composto central, para otimização das condições de gradiente. O método obtido foi o mais rápido já visto dentro da literatura consultada. A separação em linha de base foi efetuada em menos de 15 minutos, com coeficientes de variação de área entre 0,1 e 1,8%, coeficiente de correlação de 0,9993 a 0,9994 na faixa de 5 a 100 µg/mL, e limites de quantificação estimados na faixa de 0,1 a 0,21µg/mL. O método desenvolvido foi aplicado na otimização das condições de hidrólise de um extrato de Neem. A otimização foi efetuada através de metodologia de superfície de resposta, levando-se em consideração a concentração de ácido adicionada, o tempo de reação, a temperatura, e a concentração de um antioxidante (ácido ascórbico) adicionado. O resultado da otimização foi uma metodologia de hidrólise com tempo de reação igual a 5 minutos, utilizando-se 1,4 mol/L de HCI, 119°C e 500 µg/mL de ácido ascórbico. Através das metodologias de análise e de hidrólise desenvolvidas pode-se constatar a presença e quantificar no extrato de Neem os flavonóides agliconas quercetina, kaempferol e miricetina. Com o objetivo de se avaliar quais os componentes presentes em extratos vegetais são os responsáveis pelo poder antioxidante atribuído a determinadas plantas, foi montado um sistema de avaliação de poder antioxidante \"on-line\" com reação pós-coluna em HPLC (baseado na literatura) utilizando-se como \"radical livre modelo\" o ABTS. A análise da planta modelo (Neem) neste sistema mostrou que os flavonóides glicosilados identificados nas partes anteriores deste trabalho são os responsáveis pelo poder antioxidante atribuído a esta planta. De posse desta informação, e visando a obtenção de extratos para aplicações cosméticas com poder antioxidante, modelou-se a extração dos flavonóide do Neem em função da composição do solvente extrator (água, etanol , propilenoglicol e suas misturas), de acordo com um planejamento simplex centróide ampliado. Além da previsão da concentração dos princípios ativos pode-se ainda prever outras propriedades dos extratos obtidos, tais como, índice de refração e densidade, muitas vezes constituintes de especificações técnicas de acordo com as aplicações a que se destinam (cremes, xampús, etc). / At this work, separation of 18 flavonoids (9 aglycones and 9 glycosides) using High Performance Liquid Chromatography (HPLC) and Reduced Flow Micellar Electrokinetic Chromatography (RF-MEKC) were evaluated. For both techniques, pure solvents (methanol, acetonitrile and tetrahydrofuran) e their mixtures were evaluated as an approach of varying selectivity by changing mobile phase in HPLC and organic additive type in RF-MEKC. For HPLC studies using gradient elution, it was possible to guarantee the modeling for retention factor in function of organic solvent used (methanol, acetonitrile and tetrahydrofuran and theirs mixtures). It can be also confirmed, based on retention data and hierarquical clusters analysis, four different chromatographic groups with different selectivity for flavonoid aglycone, and four groups with different selectivity for glycosides. More orthogonal chromatographic systems (each one belonging to a selectivity group) were applied to Neem (Azadirachta Indica) analysis. From this study, it can be chosen the most selective mobile phase composition and optimize flavonoid glycosides separation present at Neem leaves. Applying optimized method, five major flavonoids can be identified and quantified, three quercetin glycosides (rutin, isoquercitrin and quercitrin) and two kaempferol glycosides (astragalin and nicotiflorin), at two samples from different origins (Piracicaba-SP and Silvânia-GO). For studies regarding eighteen flavonoids separation by RF-MEKC can be proved significant selectivity differences when distinct organic solvent are used as additive. Moreover, it can be noted tendencies in migration behaviour depending of solvent used and molecular structure of flavonoids. The solvent with less efficiency to f/avonoid separation is methanol. Analyzing electropherograms obtained by a design of mixtures and by criticai pairs changes observed in diverse electro/ytes, a separation method with only one criticai pair and 12 minutes run was obtained. Coefficient of variation obtained for retention factor was 1.5% and 3% for area (n=5). Developed method was applied to identify major flavonoids at model plant (Neem) and same results observed at previous work were obtained. In order to evaluate total flavonoid concentration present in a plant is a common approach to analyse extracts after acid hydrolyze (convert ali glycosides to aglycones). A method was optimized to separate 8 flavonoid aglicones by RPHPLC usually present in food and vegetal extracts to cosmetic use. Optimization was performed by a mixture factorial design to select the most selective mobile phase composition and one facto ria I design with central point to optimize gradient parameters. Developed methodology is the faster reported in literature until now. Baseline separation was achieved in less than 15 minutes, with coefficients of variation between 0.1 and 1.8%, correlation coefficient from 0,9993 to 0,9994 at 5-100 µg/mL concentration range and quantification limits from 0.1 to 0.21 µg/mL. Developed method was used to optimize hydrolize parameters for a Neem extract. Optimization was realized by a response surface methodology, having concentration of acid added, reaction time, temperature and antioxidant (ascorbic acid) concentration added as parameters. From this study was developed a hydrolyze methodology with 5 minutes of reaction time, using 1.4 mol/L HCI, 119°C and 500 µg/mL of ascorbic acid. Applying method of analysis and hydrolyze developed at Neem extracts it can be identified and quantified aglicones quercetin, kaempferol and miricetin. Aiming to evaluate which compounds in a vegetal extract have antioxidant activity credited to some plants, an on-line system with post-column reaction was built in HPLC (based on literature), using ABTS as free radical mode!. Neem analysis at this system showed that flavonoid glycosides identified before are the responsible for antioxidant activity described for this plant. Based on this information and intending to obtain vegetal extracts with antioxidant activity for cosmetic use, Neem extraction procedure was modeled in function of solvent mixture used (water, ethanol, propylene glycol and their mixtures), following a simplex centroid designo Besides the concentration of active components prediction it can also be predict other properties like refractive index and density, properties that might be included at technical specifications depending of the intended use (creams, shampoos, etc).
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