No / Budesonide is a mixture of 22R and 22S epimers. The epimeric content of budesonide was reported in both British and European pharmacopoeias to be within the range of 60-49/40-51 for R and S epimers, respectively. In this work, contribution of the two epimers to the overall infrared spectrum of budesonide has been investigated by quantum chemical calculations.

Made available in DSpace on 2015-05-14T13:00:08Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 4886781 bytes, checksum: 77da1f8dad17f8c8f812258fa8a8dd89 (MD5) Previous issue date: 2012-04-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Yangambin, a furofuran lignan isolated from Ocotea duckei Vattimo (Lauraceae) has important pharmacological activities, such as selective antagonist of platelet activating factor (PAF), cardiovascular protective effects, CNS depressant, and others. Besides, it is presented as a phytochemical marker of the specie and a promising substance for a herbal drug development. Thus, this study aimed at the development and validation of an analytical method on HPLC-DAD for simultaneous quantitative determination of yangambin and its epimer, epi-yangambin, in extracts and fractions of Ocotea duckei. For method development, a synthetic approach was made in order to obtain the lignans with high purity. The synthetic route includes a biotransformation process with coconut water peroxidase for a stereoseletive synthesis of yangambin. The chromatographic conditions for the study were: reverse phase column C-18 (250 x 4,6 mm x 5 μm), mobile phase was a mixture of water and acetonitrile (50:50) at a flow rate of 1,0 mL/min, detection at a wavelength of 215 nm. Lignans were isolated from crude ethanolic extract of the plant (leaves and stem bark) through a lignoids isolation procedure. Epimers were separated by semi-preparative HPLC-DAD to yield compounds free from impurities, being characterized by spectroscopic methods (1H and 13C NMR), infrared (IR), mass spectrometry (MS), optical rotation, circular dichroism, and thermal analysis. Validation of analytical method was performed in accordance with Resolution nº 899/2003, from ANVISA, proving to be: selective for the two epimers, linear at the concentration range of 1,0 60,0 μg/mL, precise (CV ≤ 5% ), accurate (99-101%) and robust. Application of the validated analytical method in a quantitative study allowed to determine that each 1,0g of crude ethanolic extract from stem bark and leaves presents 58,07 mg of yangambin and 88,94 mg of epi-yangambin. In total lignoids fraction, each 1,0g has 188,75 mg of yangambin and 250,02 mg of epi-yangambin. / A iangambina, lignana furofurânica isolada de Ocotea duckei Vattimo (Lauraceae), possui importantes atividades farmacológicas, como antagonista seletivo do fator de agregação plaquetária (PAF), ações protetoras cardiovasculares, depressora do SNC, dentre outras. Além disso, se apresenta como o marcador fitoquímico da espécie e é uma substância promissora para o desenvolvimento de um fitofármaco. Dessa forma, este trabalho teve como objetivo o desenvolvimento e validação de metodologia analítica em CLAE-DAD para a quantificação simultânea da iangambina e seu epímero, epi-iangambina, nos extratos e frações de Ocotea duckei. Para o desenvolvimento do método, elaborou-se uma proposta sintética para obtenção das lignanas com grau de pureza elevado. A rota sintética utiliza processo de biotransformação através da peroxidase da água de coco para a síntese da iangambina de modo estereoseletivo. As condições do método cromatográfico desenvolvido foram: coluna de fase reversa C-18 (250 x 4,6 mm x 5 μm), como fase móvel uma mistura de água e acetonitrila (50:50), a um fluxo de 1 mL/min, detecção no comprimento de onda de 215 nm. As lignanas foram isoladas do extrato etanólico bruto da planta (folhas e casca do caule) através de metodologia de isolamento para lignóides. Os epímeros foram separados através de CLAE-DAD semi-preparativa, obtendo-se os marcadores livres de impurezas, sendo caracterizados por métodos espectroscópicos (RMN de 1H e 13C), Infravermelho (IV), espectrometria de massas (EM), rotação ótica, dicroísmo circular, além de técnicas termoanalíticas. A validação da metodologia analítica foi realizada de acordo com a Resolução RE nº 899/2003 da ANVISA, mostrando-se ser: seletivo para os dois epímeros, linear na faixa de concentração de 1-60 μg/mL, preciso (CV ≤ 5%), exato (99-101%) e robusto. A aplicação da metodologia analítica num estudo quantitativo permitiu estabelecer que em 1,0g do extrato das cascas do caule e folhas tem-se cerca de 58,07 mg de iangambina e 88,94 mg de epi-iangambina. Na fração de lignóides totais, para cada 1,0g tem-se 188,75 mg de iangambina e 250,02 mg de epi-iangambina.

Vitamin-D-Mangel und -Unterversorgung haben sich zu einem großen Problem für die öffentliche Gesundheit entwickelt, das vor allem auf eine unzureichende Sonnenlichtexposition und eine begrenzte Zufuhr dieses essenziellen Nährstoffs über die Nahrung zurückzuführen ist. Diese "Pandemie" hat zu einem bemerkenswerten Anstieg der Nachfrage nach der Bestimmung des zirkulierenden 25-Hydroxyvitamin-D-Spiegels (25(OH)D) geführt. Die meisten Studien fokussieren sich in erster Linie auf 25(OH)D und betrachten es als Biomarker für den Vitamin-D-Status. Es besteht jedoch ein wachsendes Interesse an der gleichzeitigen Messung mehrerer klinisch bedeutsamer Vitamin-D-Metaboliten wie dem nativen Vitamin D, 25(OH)D-Epimeren, 1,25-Dihydroxyvitamin D und 24,25-Dihydroxyvitamin D. Die Flüssigchromatographie/Tandem-Massenspektrometrie (LC-MS/MS) gilt als "Goldstandard" für die Bestimmung des Vitamin-D-Spiegels und ermöglicht die gleichzeitige Analyse mehrerer Analyten, die für das Verständnis der physiologischen Rolle von Vitamin D und seiner klinischen Auswirkungen unerlässlich sind. In der vorliegenden Doktorarbeit soll verschiedene Aspekte der Vitamin-D-Landschaft erforschen, umfassende Einblicke geben und Herausforderungen innerhalb des Forschungsthemas besprechen. Zunächst lag der Schwerpunkt auf dem Vergleich mehrerer Derivatisierungsreagenzien für die Vitamin-D-Analyse und der Frage, wie sie sich auf die Nachweisempfindlichkeit der Methode und die chromatographische Trennung der getesteten Vitamin-D-Metaboliten auswirken. Ein zweiter Schwerpunkt lag auf der Untersuchung der Stabilität der Derivatisierungsprodukte in Serumextrakten. Der letzte Versuch war die Entwicklung einer LC-MS/MS-Methode, die die Ergebnisse der vorangegangenen Untersuchungen nutzen sollte. Als Ergebnis wurde eine neue Methode vorgestellt, die meines Wissens zum ersten Mal FMP-TS für die chemische Derivatisierung von Vitamin-D-Metaboliten verwendet. / Vitamin D deficiency has emerged as a significant public health concern, attributed largely to insufficient exposure to sunlight and limited dietary sources rich in this essential nutrient. This “pandemic” has led to a notable increase in the demand for assessing circulating levels of 25-hydroxyvitamin D (25(OH)D). Most studies have primarily focused on 25(OH)D, considered it as the vitamin D status biomarker. However, there is a growing interest in simultaneously measuring multiple clinically significant vitamin D metabolites such as the native vitamin D, 25(OH)D epimer, 1,25-dihydroxyvitamin D, 24,25-dihydroxyvitamin D and others. Thus, there is an urgent need to develop analytical methods which will be able to separate and quantify the metabolites of interest accurately and precisely. Liquid chromatography/tandem mass spectrometry (LC-MS/MS) stands out as the “gold standard” technique for vitamin D analysis. Its versatility enables simultaneous analysis of multiple analytes, thereby facilitating the generation of comprehensive vitamin D profiles essential for understanding its physiological role and clinical implications. The present doctoral thesis is designed to explore diverse aspects of vitamin D landscape, providing comprehensive insights and addressing challenges within the research topic. Initially there was a focus on comparing multiple derivatization reagents for vitamin D analysis and discovering how they impact the method’s detection sensitivity and the chromatographic separation of the tested vitamin D metabolites. Moreover, there was a second focal point on examining the stability of the derivatization products in serum extracts. The final attempt was the development of an LC-MS/MS method that would utilize the outcomes of the investigations conducted before. As a result, a new method was introduced utilizing FMP-TS for the chemical derivatization of vitamin D metabolites, a reagent which has not been reported before for vitamin D analysis.