The fibrous barley hull is the main reason for barleys low available energy relative to corn. Barley grain contains hydroxycinnamic acids (mainly ferulic acid (FA) and ñ-coumaric acid (PCA)) which are cross-linked to polysaccharides, therefore, limit cell wall degradability in the rumen. Paricle size of barley grain also affects the digestion of barley in the rumen. The objective of this study was to evaluate a set of barley varieties grown in Saskatchewan (Canada) and provided by Crop Development Center (CDC, Canada) and find a variety with low hull, FA, PCA and fiber content, while maintaining large particle size after mechanical processing, and having high nutrient availability.<p> Three studies were conducted to determine the content of barley hull, FA, PCA, neutral detergent fiber (NDF), acid detergent fiber (ADF) and acid detergent lignin (ADL) in various barley varities. Mean/median particle size of the barley grain after coarse dry-rolling was also determined. The relationships among these parameters and the digestibility of barley grain in ruminants were then assessed. Six barley varieties (AC Metcalfe, CDC Dolly, McLeod, CDC Helgason, CDC Trey and CDC Cowboy) from samples grown in three years (2003, 2004 and 2005) were evaluated in each study.<p> The first study determined the original content of barley hull, FA, PCA, NDF, ADF, ADL, and mean/median particle size of barley grain and evaluated the effects of barley variety. The results showed barley variety had a significant impact on the chemical and physical profiles of barley grain, with CDC Helgason and CDC Dolly showing relatively lower content of barley hull, FA, PCA, NDF, ADF, ADL, hemicellulose and cellulose, and moderate mean/median particle size, whereas McLeod and CDC Cowboy showed the opposite.<p> The second study involved two consecutive trials. Trial 1 was to assess differences in the in situ rumen degradability of dry matter (DM), FA, PCA, NDF, ADF and ADL at 12 and 24 h of rumen incubations. Results revealed that CDC Dolly consistently showed relatively lower rumen residues of DM, FA, PCA, NDF, ADF and ADL at 12 and 24 h, with McLeod being opposite. Barley variety displayed some effects on the digestibility of DM, FA, PCA, NDF, ADF and ADL at 12 and 24 h. Since CDC Dolly demonstrated relatively less content of hull, FA, PCA, NDF, ADF, ADL and mean/median particle size and higher rumen digestibility among the six barley varieties, while McLeod was the opposite, CDC Dolly and McLeod were selected for the third trial in order to compare differences in the rumen degradation kinetics of DM, FA and PCA. Trial 2 did not show significant differences in effective degradation of DM, FA, except for PCA. In general, CDC Dolly exhibited better degradability of DM, FA and PCA than McLeod.<p> The third study analyzed the correlation and regression between the original content of barley hull, FA, PCA, NDF, ADF, ADL and mean/median particle size in barley grain and rumen residual content of the corresponding parameters at 12 and 24 h of rumen incubation. Results showed that FA content in barley grain had a predominantly negative effect on DM degradability, while barley hull content affected the degradability of NDF and ADF.<p> In summary, the present studies show that hull and FA content in barley grain have negative effects on the degradability of barley grain in ruminants and also showed that CDC Dolly could be an ideal feed barley grain for ruminants due to its lower hull and FA content and higher rumen dry matter degradability.

The fibrous barley hull is the main reason for barleys low available energy relative to corn. Barley grain contains hydroxycinnamic acids (mainly ferulic acid (FA) and ñ-coumaric acid (PCA)) which are cross-linked to polysaccharides, therefore, limit cell wall degradability in the rumen. Paricle size of barley grain also affects the digestion of barley in the rumen. The objective of this study was to evaluate a set of barley varieties grown in Saskatchewan (Canada) and provided by Crop Development Center (CDC, Canada) and find a variety with low hull, FA, PCA and fiber content, while maintaining large particle size after mechanical processing, and having high nutrient availability.<p> Three studies were conducted to determine the content of barley hull, FA, PCA, neutral detergent fiber (NDF), acid detergent fiber (ADF) and acid detergent lignin (ADL) in various barley varities. Mean/median particle size of the barley grain after coarse dry-rolling was also determined. The relationships among these parameters and the digestibility of barley grain in ruminants were then assessed. Six barley varieties (AC Metcalfe, CDC Dolly, McLeod, CDC Helgason, CDC Trey and CDC Cowboy) from samples grown in three years (2003, 2004 and 2005) were evaluated in each study.<p> The first study determined the original content of barley hull, FA, PCA, NDF, ADF, ADL, and mean/median particle size of barley grain and evaluated the effects of barley variety. The results showed barley variety had a significant impact on the chemical and physical profiles of barley grain, with CDC Helgason and CDC Dolly showing relatively lower content of barley hull, FA, PCA, NDF, ADF, ADL, hemicellulose and cellulose, and moderate mean/median particle size, whereas McLeod and CDC Cowboy showed the opposite.<p> The second study involved two consecutive trials. Trial 1 was to assess differences in the in situ rumen degradability of dry matter (DM), FA, PCA, NDF, ADF and ADL at 12 and 24 h of rumen incubations. Results revealed that CDC Dolly consistently showed relatively lower rumen residues of DM, FA, PCA, NDF, ADF and ADL at 12 and 24 h, with McLeod being opposite. Barley variety displayed some effects on the digestibility of DM, FA, PCA, NDF, ADF and ADL at 12 and 24 h. Since CDC Dolly demonstrated relatively less content of hull, FA, PCA, NDF, ADF, ADL and mean/median particle size and higher rumen digestibility among the six barley varieties, while McLeod was the opposite, CDC Dolly and McLeod were selected for the third trial in order to compare differences in the rumen degradation kinetics of DM, FA and PCA. Trial 2 did not show significant differences in effective degradation of DM, FA, except for PCA. In general, CDC Dolly exhibited better degradability of DM, FA and PCA than McLeod.<p> The third study analyzed the correlation and regression between the original content of barley hull, FA, PCA, NDF, ADF, ADL and mean/median particle size in barley grain and rumen residual content of the corresponding parameters at 12 and 24 h of rumen incubation. Results showed that FA content in barley grain had a predominantly negative effect on DM degradability, while barley hull content affected the degradability of NDF and ADF.<p> In summary, the present studies show that hull and FA content in barley grain have negative effects on the degradability of barley grain in ruminants and also showed that CDC Dolly could be an ideal feed barley grain for ruminants due to its lower hull and FA content and higher rumen dry matter degradability.

Heritage apple cultivars for cider-making are often distinguished by a high concentration of tannins (phenolic compounds), and/or acid. The phenolic content of some cider apples far exceeds that of white wine, however most cider fermentation practices are directly taken from white winemaking, not accounting for effects of high concentrations of phenolic compounds on yeast fermentation. The objective of this study was to determine the impact of ferulic acid, p-coumaric acid, and chlorogenic acid—at concentrations reported in apples—and their interactions with yeast assimilable nitrogen (YAN) on fermentation kinetics and cider aroma. Our hypothesis was that the phenolic compounds present in high-tannin cider apples would negatively impact fermentation kinetics, but not alter the aroma, and that added YAN would reduce these effects. Ferulic acid negatively affected fermentation performance (p < 0.05), but p-coumaric acid and chlorogenic acid did not. p-Coumaric acid led to the greatest changes in cider aroma. Differences were also detected for different concentrations of ferulic acid. Chlorogenic acid did not affect aroma. Yeast strain influenced fermentation performance and cider aroma. Finally, addition of exogenous YAN improved fermentation performance for the low concentration ferulic acid condition, but not for the high concentration. Adding YAN also changed cider aroma in the presence of p-coumaric acid. / Master of Science in Life Sciences / Hard cider is increasingly popular in the United States. Heritage apple cultivars are traditional cider apples, often distinguished by a high concentration of tannins (phenolic compounds), and/or acid, unlike dessert apples that are typically higher in sugar than tannins. While the phenolic content of some cider apples far exceeds that of white wine, most cider fermentation practices are directly taken from white winemaking, and do not account for the effects of high concentrations of phenolic compounds on fermentation performance. The objective of this study was to determine whether three phenolic compounds—ferulic acid, p-coumaric acid, and chlorogenic acid—at concentrations reported in apples, would inhibit fermentation or alter the aroma of the cider, and if adding yeast assimilable nitrogen (YAN), a yeast nutrient, would reduce these effects. Our hypotheses were that the phenolic compounds present in high-tannin cider apples would negatively impact fermentation performance, but not alter the aroma, and that added YAN would reduce these effects. Ferulic acid negatively affected fermentation performance (p < 0.05), but p-coumaric acid and chlorogenic acid did not. Addition of nitrogen improved fermentation performance for the low concentration ferulic acid condition, but not for the high concentration. p-Coumaric acid led to the greatest changes in cider aroma, with differences in aroma also detected for ferulic acid ciders. Chlorogenic acid did not affect aroma or fermentation performance. Yeast strain and YAN addition also influenced fermentation performance and aroma.

4-Ethylphenol (4-EP) is a malodorant of swine waste and is derived from a component of lignin called p-coumaric acid (p-CA). The production of 4-EP from lignin in swine waste is untested. Additionally, the effect of Fe (III) on 4-EP levels is unknown. Four experiments were performed to determine if Lactobacillus sp. pep8 cultures, as well as enriched swine lagoon slurries, could liberate p-CA from lignin and convert p-CA to 4-EP. Furthermore, it was tested if the addition of Fe (III) influences the conversion of p- CA to 4-EP. Experiment 1 tested Lactobacillus sp. pep8 cultures to determine if the addition of 10 mM Fe (III) and 0.2% sulphite lignin to Lactobacillussp. pep8 cultures would stimulate production of 4-EP. Experiment 2 tested the effect of 0.2% sulphite lignin and 10 mM Fe (III) on 4-EP production in the presence of enriched swine lagoon slurries. On day 0 there was no detectable 4‐EP, for either 0.2% sulphite lignin addition or the 10 mmol l‐1 Fe (III) additions. Experiment 3 tested alternative forms of lignin, including 0.2% sulphite, indulin, or sigma lignin as potential source compounds for 4-EP production in enriched swine lagoon slurries. 4-EP produced in all three conditions are likely endogenous to the lagoon slurry additions. Experiment 4 was designed to measure the degradation of exogenous 4-EP with varying final concentrations of 4-EP in enriched swine lagoon slurries. Data in Figure 7 indicate immediate degradation of 4-EP by day 5, however, by day 7 synthesis of 4-EP occurred until day 14 where 4-EP levels remained in a steady state. Our results suggest that when both Lactobacillus sp. pep8 cultures and enriched swine lagoons are supplemented with p-CA, 4-EP is produced indicating that p-CA serves as a source of 4-EP. However, when supplemented with Fe (III) and/or sulphite, indulin, or sigma lignin, 4-EP production was not stimulated. This data indicates that, 4- EP production is not enhanced by the presence of Fe (III) in either Lactobacillus sp. pep8 cultures or in enriched swine lagoon slurries. Furthermore, lignin did not serve as a source of 4-EP.

Orientador: Marcos Antonio Correa / Resumo: O ácido p-cumárico (APC) é um composto fenólico não flavonóide que apresenta segurança e eficácia bem descritas na ciência dos alimentos devido suas propriedades antioxidante, antitumoral, anti-inflamatória, antimicrobiana e por apresentar baixa toxicidade oral. Essas propriedades são pouco exploradas em produtos para cuidados com a pele. O objetivo do presente estudo foi avaliar de forma in vitro a segurança e a eficácia do APC e incorporá-lo em emulsão cosmética óleo em água (O/A). Nossos estudos in vitro e alternativos ao uso de animais foram conduzidos para verificar a citotoxicidade do APC em células de queratinócitos (HaCat) e fibroblastos humanos (HDFa), o potencial de irritação ocular (HET-CAM) e a atividade antimicrobiana em diferentes cepas. Emulsões contendo APC foram submetidas ao estudo de estabilidade e de liberação, retenção e permeação cutâneas. Resultados: o IC50 do APC em células HaCat e HDFa foi 693,57 e 2.814,75 μg/mL, respectivamente. Além disso 100 μg de APC foram levemente tóxicos em membrana corioalantóica de ovo embrionado. As concentrações bactericidas mínimas em cepas de E. coli, S. aureus, P. aeruginosa e S. epidermidis, foram 3.000 μg/mL, 1.875 μg/mL, 3.750 μg/mL e 3.750 μg/mL, respectivamente. Emulsões contendo diferentes concentrações de APC foram estáveis durante o período de estabilidade acelerada. Após 12 horas 91,08 ± 0,03% do APC foi liberado da formulação e após 24 horas, 0,52 ± 0,18% e 0,94 ± 0,37% ficaram retidos no estrato córneo e na e... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: p-Coumaric acid (PCA) is a non flavonoid phenolic compound whose safety and efficacy widely described in food science papers due to antioxidant, antitumor, anti inflammatory properties, and low oral toxicity. These characteristics are poorly or no described when PCA is added to skin care products. The aim of the present study was to evaluate the safety and efficacy of PCA through in vitro assays, and then add it in oil in water cosmetic emulsion (O/W). Our in vitro trials, without animal use were conducted to predict PCA´s cytotoxicity in HaCat and HDFa cells, ophthalmic irritation potential, and antibacterial activity against differents strains. Emulsions with PCA has been tested to predict shelf life, dermal absorption, and dermal retention. Results: PCA´s IC50 in HaCat and HDFa cells was 693.57 and 2,814.75 μg.mL-1, respectively. In addition 100 μg of APC were slightly toxic in chorioallantoic membrane assay. The minimum bactericidal concentrations against E. coli, S. aureus, P. aeruginosa and S. epidermidis strains were 3,000 μg.mL-1, 1,875 μg.mL-1, 3,750 μg.mL-1, and 3,750 μg.mL-1 , respectively. Emulsions containing 0.04%, 0.06%, 0.08% and 0.10% of PCA were stable during self life testing for ninety days. After 12 hours 91.08 ± 0.03% of PCA was released of the emultion, and after 24 hours 0.52 ± 0.18% and 0.94 ± 0.37%, were present in stratum corneum and epidermis/dermis, respectively. Our findings indicate that PCA concentrations would be effective and safe to use in c... (Complete abstract click electronic access below) / Mestre

Most of the biological systems in nature are sustained by molecular self-assemblies which are the finest examples of supramolecular architectures. Non-covalent interactions are key concepts which govern these molecular assemblies. Inspired by these examples crystal engineering emerged as an important tool in supramolecular chemistry which aids in the invention of new molecular structures with desired properties. Understanding of how the molecules interact at the molecular levels enables one to rationally design novel solid forms with modulated physicochemical properties. This feature of crystal engineering has heightened its position in materials chemistry and is currently one of the most well studied fields for generating novel compounds with pre-defined composition and supramolecular architectures. One such class of compounds that has immensely attracted the scientific community and is under continuous study for wider applications is cocrystals. The applications include various interdisciplinary fields such as pharmaceutics, catalysis, organic conductors, explosives etc. Distinctly on the other side, cocrystals also provide a means to discover new supramolecular synthons which is the ultimate key to molecular assembly. Many robust supramolecular synthons have been discovered and hierarchies are also being developed which can serves as a design tool for cocrystal synthesis. The Cambridge Structural Database (CSD) is an important accessory in determining the robustness of a supramolecular synthon but, this does not preclude us from discovering new synthons. The work presented here explores new persistent supramolecular synthons in polyphenols utilizing the basic concepts of crystal engineering and the CSD statistical analysis. This contribution also includes the implementation of cocrystallization for various categories of compounds which includes nutraceuticals, pharmaceuticals and ionic salts for the design and synthesis of molecular and ionic cocrystals. Chapter 1 highlights how supramolecular synthon approach can be used to design and synthesize multi-component crystals, namely, cocrystals. The role of the CSD and its importance in crystal engineering has also been discussed. Chapters 2 and 3 focus on new persistent supramolecular synthons in the context of nutraceuticals. The cocrystals isolated in the study are also compared with the existing cocrystals in the CSD supramolecularly in terms of synthon formation. These persistent supramolecular synthons are helpful in developing hierarchies which could be utilized and applied to similar and analogous compounds. The main feature of Chapter 4 is expanding the field of cocrystallization by studying the properties of cocrystals. Some of the properties which have been examined here include effects of cocrystallization on solubility and correlations between the solubility of cocrystal with cocrystal former (CCF) and melting point of the cocrystal. The extension of cocrystals to the active pharmaceutical ingredients (APIs) has been explored in the context of pharmaceutical cocrystals by selecting a BCS class IV drug, hydrochlorothiazide in Chapter 5. Chapter 6 highlights the hybridization of organic and inorganic components for the synthesis of ionic cocrystals and is exemplified by considering lithium salts with achiral and homochiral amino acids for the generation of 1:1 and 1:2 cocrystals.

Tyrimo tikslas: sumodeliuoti puskietes sistemas su bičių produktais ir įvertinti jų kokybę biofarmaciniais tyrimais. Uždaviniai: 1. Atrinkti tinkamą propolio, kaip veikliosios medžiagos, formą įvedimui į modeliuojamas puskietes sistemas; 2. Pagaminti stabilias puskietes formas su bičių produktais (bičių vašku, propoliu, medumi) ir įvertinti jų kokybę, tiriant šiuos parametrus: vienalytiškumas, pH reikšmė, antioksidacinis aktyvumas; 3. Atlikti puskiečių sistemų su bičių produktais biofarmacinį vertinimą ir identifikuoti veiksnius, darančius įtaką bendro fenolinių junginių kiekio atpalaidavimui; 4. Įvertinti puskiečių formų su bičių produktais kokybę, remiantis atliktais veikliųjų junginių (fenolinių rūgščių, vanilino) atpalaidavimo in vitro tyrimo rezultatais. Bičių produktų puskiečių sistemų analizei pasirinktas fenolinių junginių nustatymas spektrofotometriniu ir ESC metodais bei spektrofotometrinis metodas laisvųjų DPPH• radikalų surišimo aktyvumui pagal troloksą nustatyti. Vaisto formų kokybė vertinta nustatant pH reikšmę ir atliekant veikliųjų junginių atpalaidavimo tyrimą in vitro. Į puskietes sistemas veiklioji medžiaga (propolis) įterpiama tirštojo arba skystojo (gelyje) ekstrakto forma. Stabilios I grupės sistemos pagaminamos tada, kai bičių vaško ir saulėgrąžų aliejaus santykis ne didesnis kaip 1:5. Bičių vaškas ir medus daro įtaką atpalaiduojamam veikliųjų junginių kiekiui – didėjant vaško kiekiui, veikliųjų junginių iš sistemų atsipalaiduoja mažesnis kiekis, o iš... [toliau žr. visą tekstą] / The aim of work: Formulate and analyze semisolid preparations with bees products and evaluate their quality by biopharmaceutical researches. Main tasks: 1. Select the proper form of propolis as an active material in formulated semisolid systems; 2. Formulate stable semisolid systems with bees products (beeswax, propolis, honey) and evaluate their quality by investigation of these parameters: homogenity, pH value, antioxidant activity; 3. Perform biopharmaceutical evaluation of semisolid systems with bees products and identify the factors that influence release the total amount of phenolic compounds; 4. Evaluate quality of semisolid systems with bees products based on active compounds (phenolic acids, vanillin) in vitro release test results. Semisolid systems with bees products are analyzed by using spectrophotometric and capillary HPLC methods for analysis of total phenolic compounds measuring, DPPH• free radical scavenging activity measuring according to trolox. Quality of the systems was evaluated by measuring pH values and by testing release of active compounds in vitro. The active substance propolis is inserted into semisolid systems by thick or liquid (in gels) extract form. Stable system of the Group I semisolid systems is formulated when beeswax and sunflower oil ratio is less than 1:5. Beeswax and honey in systems affect the release of active compounds - increasing the wax content, the active compounds release of the systems is reduced, adequate quantity of honey... [to full text]

Orientadoresr: Daniele Ribeiro de Araujo, Eneida de Paula / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-21T08:56:23Z (GMT). No. of bitstreams: 1 Guilherme_VivianeAparecida_M.pdf: 3020599 bytes, checksum: ce06e2f6da5344128ba90d18a9b569f2 (MD5) Previous issue date: 2012 / Resumo: Compostos anti-inflamatórios não esteroídais (AINEs) são amplamente utilizados para o combate da inflamação, mas frequentemente acarretam efeitos adversos que impedem a continuação do tratamento. Atualmente, a terapia a base de plantas tem sido bastante empregada a fim de desenvolver novos fármacos que apresentem eficácia analgésica e anti-inflamatória. Nesse contexto, a utilização de sistemas carreadores, como as nanopartículas de hidróxidos duplos lamelares (HDL), visa melhorar propriedades biofarmacêuticas e farmacológicas dos compostos nelas intercalados. Dessa forma, este estudo teve como objetivo caracterizar a interação entre o ácido coumárico (AC), um composto fenólico extraído de plantas, e hidróxido duplo lamelares (HDL), nanopartículas inorgânicas, considerando parâmetros como a cinética de liberação in vitro, a citotoxicidade e as atividades analgésica-antiinflamatória em relação ao fármaco não intercalado. Os ensaios de liberação in vitro foram realizados utilizando uma célula de difusão vertical (com membrana de acetato de celulose, MWCO 1000 Da) e a viabilidade celular avaliada em células 3T3 pelo teste de incorporação do vermelho neutro (VN). Os testes farmacológicos realizados em camundongos swiss foram: a determinação do número de contorções abdominais, teste tail-flick, ensaios de peritonite e teste de formalina para o composto livre e intercalado nas concentrações de 10, 20 ou 30 mg/kg. Os ensaios de liberação in vitro mostraram que a intercalação reduziu significativamente a constante de liberação (Klib) do AC, em relação ao fármaco livre, sendo os valores de Klib de 41,6 ± 1,5 %.h-1 e 32,4 ± 1,5 %.h-1 , para o AC e AC-HDL, respectivamente. A viabilidade celular foi reduzida apenas em concentrações mais elevadas de AC e AC-HDL (10 e 12,5 mM). Porém, mesmo nestas concentrações foi observada porcentagem de viabilidade celular maior que 50%. Por fim, a avaliação farmacológica apontou o AC-HDL como um sistema de liberação com atividades analgésica e anti-inflamatória mais pronunciadas do que as observadas para anti-inflamatórios não esteroidais como a indometacina (p< 0,001). Esses efeitos foram obtidos para teste de tail-flick, quando o AC foi intercalado em HDL aumentou a duração da analgesia (~ 1,7 vezes) quando comparado com o grupo de controle indometacina. Assim, os resultados indicam que o AC intercalado em nanopartículas inorgânicas de HDL apresentou uma taxa de liberação lenta e também induziu uma atividade analgésica - anti-inflamatória, possivelmente, por um mecanismo semelhante ao observado para um anti-inflamatório não esteroidais como a indometacina / Abstract: Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used to avoid inflammation, but these compounds can evoke some side effects, considering these as an important limitation to the treatment. Currently, the plant-based therapy has been widely employed to develop new drugs which have analgesic and anti-inflammatory pharmacological activities. In this context, the use of inorganic nanoparticles is important to improve biopharmaceutical and pharmacological properties of the intercalated molecules. Thus, this study aimed to characterize the interaction between coumaric acid (CA), a phenolic compound extracted from plants, and layered double hydroxide (LDH), inorganic nanoparticles, considering parameters such as the in vitro release kinetics cytotoxicity and analgesic-antiinflammatory activities compared to the non-intercalated-CA. In vitro release tests were performed using a vertical diffusion cell (with cellulose acetate membrane, MWCO 1000 Da) and cell viability assessed in 3T3 cells by the neutral red (NR) uptake test. Pharmacological assays were carried in Swiss mice out in order to determine the number of writhings, tail-flick test, peritonitis test and formalin test for the free compound at three different concentrations (10, 20 or 30 mg/kg). In vitro release tests showed that the release constant (Krel) value was significantly reduced when compared to the non-intercalated CA (Krel values of 41.6 ± 1.5 %.h-1 and 32.4 ± 1.5 %.h-1 for the free CA and CA-HDL, respectively). Cell viability was reduced only at higher concentrations (10 and 12.5 mM) of CA and CA-HDL. However, even at these concentrations the percentage of cell viability was more than 50%. Finally, the pharmacological evaluation reveal the CA-HDL as a drug-release system with more pronounced analgesic-antiinflammatory effects than those observed for classic NSAIDs, such as indomethacin (p <0.001). Those effects were obtained, specially, for tail-flick test, when the treatment with CA-LDH increased the duration of analgesia (~ 1.7 times), when compared with the control group (indomethacin). Thus, the results pointed out that the system CA-LDH showed a slow release rate and also induced in vivo analgesic-anti-inflammatory activities, possibly using similar mechanisms to that observed for classic non-steroidal anti-inflammatory drugs, such as indomethacin / Mestrado / Bioquimica / Mestre em Biologia Funcional e Molecular

A intercalação de espécies com atividade farmacológica em Hidróxidos Duplos Lamelares (HDLs) vem sendo explorada com uma maior freqüência não apenas pelo fato da matriz ser biocompatível, mas também por outros efeitos reportados em estudos recentes como: (i) possível liberação sustentada da droga mediada por alterações no pH, (ii) aumento da solubilidade de substâncias pouco solúveis em água, (iii) aumento da estabilidade química de substâncias frente à luz, ao calor, à umidade, ao oxigênio molecular etc. O principal objetivo deste trabalho é a intercalação de substâncias de interesse medicinal (em suas formas aniônicas) como a pravastatina, Prav (ação anti-hiperlipidêmica) e os antioxidantes ácidos coumárico, Cou, e lipóico, Lip, em HDLs de magnésio-alumínio e zinco-alumínio através da coprecipitação, utilizando diferentes condições experimentais (pH, tratamento pós-síntese e relação molar ânion/Al3+). As amostras sólidas foram caracterizadas por análise elementar (CHN e metais), difratometria de raios X, métodos espectroscópicos (vibracional no infravermelho, Raman e ressonância magnética nuclear de 13C no estado sólido), análise térmica, microscopia eletrônica de varredura, medidas de tamanho de partícula e de potencial Zeta. A ação farmacológica dos HDLs de composição Mg2Al foi investigada através do teste biológico com o corante Sulforodamina B na linhagem celular de melanoma A-375. A utilização de diferentes proporções molares Pravastatina/Al3+ (1, 2 e 3) não alterou significativamente a quantidade de substância orgânica presente no material. O HDL de composição lamelar Zn2+/Al3+ apresentou maior cristalinidade e maior quantidade de orgânico (52 % m/m) que a matriz Mg2+/Al3+ (32 % m/m). A Pravastatina forma uma bicamada na região interlamelar. Os valores de tamanho médio de partícula e do potencial Zeta encontrado para o material Mg2Al1Prav (razão molar Prav/Al3+= 1) são 126 nm e + 22,9 mV, respectivamente. A Pravastatina e o material de Mg2Al1Prav diminuíram o crescimento celular da linhagem A-375 em até 20 % após tratamento. A composição dos HDLs intercalados com íons coumarato é dependente da razão molar Cou/Al3+ empregada na síntese. Observou-se novamente que o emprego da composição lamelar Zn2+/Al3+ contribui para a obtenção de material com maior cristalinidade e quantidade de orgânico intercalado (36 % m/m) comparado ao análogo de Mg2+/Al3+ (32 % m/m). Os ânions orgânicos na forma bivalente, constatados pelos resultados obtidos no espectro Raman, assumem um arranjo de monocamada com inclinação do esqueleto carbônico em relação às lamelas. Dados de análise elementar e análise térmica sugerem que o íon orgânico está enxertado/coordenado às lamelas do HDL. A suspensão aquosa do material Zn2Al3Cou apresenta estabilidade moderada frente à aglomeração (potencial Zeta igual a + 39,6 mV) e tamanho médio de partícula de 226 nm. No caso dos HDLs-Lip, observou-se que o excesso de íons lipoato na síntese, assim como o tempo de agitação pós-síntese , não alterou a cristalinidade e a quantidade de orgânico (39 % m/m) no material. Os ânions orgânicos devem se organizar na forma de uma bicamada interdigitada na região interlamelar. A suspensão aquosa do material Mg2Al1Lip apresenta estabilidade moderada frente à aglomeração (potencial Zeta igual a + 34,3 mV) e tamanho médio de partícula de 110 nm. / The intercalation of species with pharmacological activity in Layered Double Hydroxide (LDHs) has been explored frequently not only because it is biocompatible, but also by others effects as reported in recent studies: (i) sustained release of the drug by changes in pH, (ii) increasing the solubility of poorly water soluble substances, (iii) increase of the chemical stability of substance against light, heat, moisture, molecular oxygen etc. The main goal of this work is the intercalation of substances of therapeutic interest (in their anionic forms) as Pravastatin, Prav (antihyperlipidemic), and the antioxidants coumaric acid, Cou, and lipoic acid, Lip, in LDHs of magnesium-aluminum and zinc-aluminum by coprecipitation using different experimental conditions (pH, post-synthesis treatment and molar ratio anion/Al3+). Solid samples were characterized by elemental analysis (CHN and metals), X-ray diffraction, spectroscopic methods (Vibrational Infrared, Raman and Nuclear Magnetic Resonance of 13C in solid state), thermal analysis, scanning electron microscopy, measurements of particle size and Zeta potential. The pharmacological activity of LDHs of Mg2Al composition was investigated by the biological assay with the dye Sulforhodamine B in the melanoma cell line A-375. The usage of different molar ratio Pravastatin/Al3+ (1, 2 and 3) did not significantly alter the amount of organic substance present in the inorganic carrier. The LDH of composition Zn2+/Al3+ promoted an increase in the crystallinity of the material and in the amount of organic (52 % w/w) compared to the Mg2+/Al3+ material (32 % w/w). Pravastatin forms a bilayer into the interlayer region. The mean particle size and Zeta potential of the Mg2Al1Prav (molar ratio Prav- /Al3+= 1) are 126 nm and + 22.9 mV, respectively. The Pravastatin and the material Mg2Al1Prav decreased cell growth of strain A-375 melanoma by 20 % after treatment. The composition of the LDH intercalated with coumarate ions is dependent of the molar ratio anion/Al3+ used in the synthesis. The composition Zn2+/Al3+ promoted an increase in the crystallinity and amount of the organic (36 % w/w) compared to Mg2+/Al3+ material (32 % w/w). The organic divalent anions, visualized by the Raman spectra results, assume a monolayer arrangement with the inclination of the carboxylate group related to the layer. Elemental and thermal analysis suggest the grafting/coordination of the organic ions into the layers of LDH. The aqueous suspension of the Zn2Al3Cou shows moderate stability against agglomeration (Zeta potential value + 39.6 mV) and particle size of 226 nm. In the case of LDHs-Lip, it was observed that excess of lipoate ions from the synthesis, as well as, the stirring time pos-synthesis did not alter the crystallinity and the organic amount (39 % w/w) material. The organic anions are organized in an interdigitated bilayer into the interlayer region. The aqueous suspension of the Mg2Al1Lip material shows moderate stability against the agglomeration (zeta potential value equal to + 34.3 mV) and a particle size of about 110 nm.

A intercalação de espécies com atividade farmacológica em Hidróxidos Duplos Lamelares (HDLs) vem sendo explorada com uma maior freqüência não apenas pelo fato da matriz ser biocompatível, mas também por outros efeitos reportados em estudos recentes como: (i) possível liberação sustentada da droga mediada por alterações no pH, (ii) aumento da solubilidade de substâncias pouco solúveis em água, (iii) aumento da estabilidade química de substâncias frente à luz, ao calor, à umidade, ao oxigênio molecular etc. O principal objetivo deste trabalho é a intercalação de substâncias de interesse medicinal (em suas formas aniônicas) como a pravastatina, Prav (ação anti-hiperlipidêmica) e os antioxidantes ácidos coumárico, Cou, e lipóico, Lip, em HDLs de magnésio-alumínio e zinco-alumínio através da coprecipitação, utilizando diferentes condições experimentais (pH, tratamento pós-síntese e relação molar ânion/Al3+). As amostras sólidas foram caracterizadas por análise elementar (CHN e metais), difratometria de raios X, métodos espectroscópicos (vibracional no infravermelho, Raman e ressonância magnética nuclear de 13C no estado sólido), análise térmica, microscopia eletrônica de varredura, medidas de tamanho de partícula e de potencial Zeta. A ação farmacológica dos HDLs de composição Mg2Al foi investigada através do teste biológico com o corante Sulforodamina B na linhagem celular de melanoma A-375. A utilização de diferentes proporções molares Pravastatina/Al3+ (1, 2 e 3) não alterou significativamente a quantidade de substância orgânica presente no material. O HDL de composição lamelar Zn2+/Al3+ apresentou maior cristalinidade e maior quantidade de orgânico (52 % m/m) que a matriz Mg2+/Al3+ (32 % m/m). A Pravastatina forma uma bicamada na região interlamelar. Os valores de tamanho médio de partícula e do potencial Zeta encontrado para o material Mg2Al1Prav (razão molar Prav/Al3+= 1) são 126 nm e + 22,9 mV, respectivamente. A Pravastatina e o material de Mg2Al1Prav diminuíram o crescimento celular da linhagem A-375 em até 20 % após tratamento. A composição dos HDLs intercalados com íons coumarato é dependente da razão molar Cou/Al3+ empregada na síntese. Observou-se novamente que o emprego da composição lamelar Zn2+/Al3+ contribui para a obtenção de material com maior cristalinidade e quantidade de orgânico intercalado (36 % m/m) comparado ao análogo de Mg2+/Al3+ (32 % m/m). Os ânions orgânicos na forma bivalente, constatados pelos resultados obtidos no espectro Raman, assumem um arranjo de monocamada com inclinação do esqueleto carbônico em relação às lamelas. Dados de análise elementar e análise térmica sugerem que o íon orgânico está enxertado/coordenado às lamelas do HDL. A suspensão aquosa do material Zn2Al3Cou apresenta estabilidade moderada frente à aglomeração (potencial Zeta igual a + 39,6 mV) e tamanho médio de partícula de 226 nm. No caso dos HDLs-Lip, observou-se que o excesso de íons lipoato na síntese, assim como o tempo de agitação pós-síntese , não alterou a cristalinidade e a quantidade de orgânico (39 % m/m) no material. Os ânions orgânicos devem se organizar na forma de uma bicamada interdigitada na região interlamelar. A suspensão aquosa do material Mg2Al1Lip apresenta estabilidade moderada frente à aglomeração (potencial Zeta igual a + 34,3 mV) e tamanho médio de partícula de 110 nm. / The intercalation of species with pharmacological activity in Layered Double Hydroxide (LDHs) has been explored frequently not only because it is biocompatible, but also by others effects as reported in recent studies: (i) sustained release of the drug by changes in pH, (ii) increasing the solubility of poorly water soluble substances, (iii) increase of the chemical stability of substance against light, heat, moisture, molecular oxygen etc. The main goal of this work is the intercalation of substances of therapeutic interest (in their anionic forms) as Pravastatin, Prav (antihyperlipidemic), and the antioxidants coumaric acid, Cou, and lipoic acid, Lip, in LDHs of magnesium-aluminum and zinc-aluminum by coprecipitation using different experimental conditions (pH, post-synthesis treatment and molar ratio anion/Al3+). Solid samples were characterized by elemental analysis (CHN and metals), X-ray diffraction, spectroscopic methods (Vibrational Infrared, Raman and Nuclear Magnetic Resonance of 13C in solid state), thermal analysis, scanning electron microscopy, measurements of particle size and Zeta potential. The pharmacological activity of LDHs of Mg2Al composition was investigated by the biological assay with the dye Sulforhodamine B in the melanoma cell line A-375. The usage of different molar ratio Pravastatin/Al3+ (1, 2 and 3) did not significantly alter the amount of organic substance present in the inorganic carrier. The LDH of composition Zn2+/Al3+ promoted an increase in the crystallinity of the material and in the amount of organic (52 % w/w) compared to the Mg2+/Al3+ material (32 % w/w). Pravastatin forms a bilayer into the interlayer region. The mean particle size and Zeta potential of the Mg2Al1Prav (molar ratio Prav- /Al3+= 1) are 126 nm and + 22.9 mV, respectively. The Pravastatin and the material Mg2Al1Prav decreased cell growth of strain A-375 melanoma by 20 % after treatment. The composition of the LDH intercalated with coumarate ions is dependent of the molar ratio anion/Al3+ used in the synthesis. The composition Zn2+/Al3+ promoted an increase in the crystallinity and amount of the organic (36 % w/w) compared to Mg2+/Al3+ material (32 % w/w). The organic divalent anions, visualized by the Raman spectra results, assume a monolayer arrangement with the inclination of the carboxylate group related to the layer. Elemental and thermal analysis suggest the grafting/coordination of the organic ions into the layers of LDH. The aqueous suspension of the Zn2Al3Cou shows moderate stability against agglomeration (Zeta potential value + 39.6 mV) and particle size of 226 nm. In the case of LDHs-Lip, it was observed that excess of lipoate ions from the synthesis, as well as, the stirring time pos-synthesis did not alter the crystallinity and the organic amount (39 % w/w) material. The organic anions are organized in an interdigitated bilayer into the interlayer region. The aqueous suspension of the Mg2Al1Lip material shows moderate stability against the agglomeration (zeta potential value equal to + 34.3 mV) and a particle size of about 110 nm.