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Navegando Mestrado em Ciências Farmacêuticas (FF) por Por Orientador "BARA, Maria Teresa Freitas"
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Item Estudo da composição química, de atividades biológicas e microencapsulação do óleo essencial dos frutos de Pterodon emarginatus Vogel - Fabaceae ("sucupira")(Universidade Federal de Goiás, 2012-02-28) ALVES, Suzana Ferreira; CONCEIÇÃO, Edemilson Cardoso da; http://lattes.cnpq.br/7193007113950510; BARA, Maria Teresa Freitas; http://lattes.cnpq.br/3914164125498267The present work aimed to do the study of chemical variability, evaluating biological activities and microencapsulate the essential oil of the fruits of Pterodon emarginatus Vogel (Fabaceae). Chapter 1 presents the study of the chemical variability of essential oil from fruits of sucupira of 11 individual of the Brazilian savanna, collected from five different populations, and were identified as its chemical composition by gas chromatography coupled to mass spectrometry (GC/MS), used for data analysis to multivariate statistical tool that indicated compounds β caryophyllene and α copaene as the main discriminant of the samples studied, both of which are of greater significance. Chapter 2 describes the antimicrobial activity and chemical composition of total oil of the fruits of sucupira, four samples being analyzed, three of the city of Jussara GO and one of region of Jaciara MT. The oils tested showed good antimicrobial activity against bacteria Gram positive (Gram (+)) and moderate activity against Enterobacter aerogenes ATCC 13048. Chapter 3 describes the process of obtaining microcapsules containg essential oil (OE) of sucupira employing the technique of spray drying and the development and validation methodology for quantification of the compound β caryophyllene. Were used gum Arabic and maltodextrin as wall material and prepared five different dispersions (Emulsion 1-E1, Emulsion 2-E2, Emulsion 3-E3, Emulsion 4-E4 and Emulsion 5-E5) which then were atomized in spray dryer. The results show that the drying condition most appropriate was sprinkler beak of 1,2 mm of diameter, power flow the emulsion in system of 4 mL/min, inlet temperature of 160ºC, air flow of 40 L/min e and pressure of 60 psi. Among the emulsions, E2 was standardized with adequate proportion of essential oil and wall materials, by presenting microcapsules (MOE) for having thick-walled, with a pronounced retention of essential oil, spherical morphology and low hygroscopicity. The method developed and validated proved to be linear, precise, acuurate and robust. Chapter 4 presents the evaluation of biological activities of antimicrobial essential oil of PES-01 employing the technique of broth microdilution and antinociceptive and anti-inflamatory activities performed to the microcapsules with the essential oil. The models of pain induced by formalin and hot plate were used to assess the antinociceptive activity and the model of carrageenan-induced pleurisy and Evans blue for evaluation of anti-inflamatory activity. The OE has weak antimicrobial activity (500 μg mL-1) against bacteria Gram (+) and against the fungi of the genus Cryptococcus was inactive against S. aureus ATCC 25923, against bacteria Gram (-) and against the fungi of the genus Candida. OE reduced by 61,54% (300 mg kg-¹) of the reactivity to pain. The MOE reduced 40,87% (1,0 g kg-¹) and 41,57% (2,0 g kg-¹) in first phase of the test formalin-induced pain, suggesting, anti-nociceptive activity. The 2nd phase of the test, the essential oil inhibited 52,35% of the pain related to inflammatory mediators and the microcapsules decreased from 25,86% - 55,60% of the pain. MOE at a dose of 1,0 g kg-¹ showed anti-nociceptive activity in hot plate model, suggestion central analgesic activity. In the arrageenaninduced pleurisy the MOE reduced 25,44% of the complete migration of leukocytes and significantly decreased the concentration of Evans blue in 24,18%, which demonstrates important anti-inflamatory activity.Item Estudo fitoquímico e avaliações da toxicidade aguda e atividades biológicas da raiz do vetiver (Vetiveria zizanioides L. Nash) Poaceae(Universidade Federal de Goiás, 2009-03-23) BARROS, Gilvana Cristina de; BARA, Maria Teresa Freitas; http://lattes.cnpq.br/3914164125498267The use of medicinal plants in controlling the diseases has shown its potential for the discovery of new medicines. Among the plant species used in the Hospital de Medicina Alternativa (HMA / SES-GO) highlights the vetiver (Vetiveria zizanioides L. Nash), an aromatic plant of the family Poaceae, used as a mild anti-hypertensive and diuretic. In this work it was investigated the chemical composition of the hydroalcoholic extract of vetiver, the freeze-dried aqueous extract and the essential oil from the roots of V. zizanioides. In the hydroalcoholic extract were found terpenoids compounds and / or saponins, flavonoids and phenolic acid. In the freeze-dried aqueous extract was observed the presence of flavonoids and phenols. In the essential oil 38 compounds were identified, and khusimol, palustrol, khusimona, zizanona and epi-α-vetivona were the major constituents. In the assessment of the acute toxicity of the hydroalcoholic extract and the freeze-dried aqueous extract of root of V. zizanioides, it was concluded that these extracts showed no toxicity at a dose of 2000 mg / kg. The freeze-dried aqueous extract of the roots of V. zizainoides showed no statistically significant diuretic action in the doses tested. In the investigation of antimicrobial activity, the essential oil of the root of V. zizanioides showed potent activity to inhibiting the growth of Staphylococcus aureus with MIC of 0.15 mg / mL. The crude ethanolic extract showed an antimicrobial activity with MIC of 0.19 mg / mL for Bacillus cereus and Micrococcus roseus and 0.39 mg / mL for Micrococcus luteus, S. aureus and Bacillus subtilis. The hexane fraction showed stronger antimicrobial activity against strains of M. luteus, M. roseus, S. aureus and B. cereus. This study could prove that the essential oil, the crude ethanol extract and hexane fraction from roots of V. zizanioides showed antimicrobial potential.Item APLICAÇÃO DO ENSAIO DE DISSOLUÇÃO NA AVALIAÇÃO DA QUALIDADE DE MEDICAMENTOS FITOTERÁPICOS À BASE DE Passiflora sp(Universidade Federal de Goiás, 2008-10-29) COSTA, Ane Rosalina Trento; BARA, Maria Teresa Freitas; http://lattes.cnpq.br/3914164125498267Phytomedicines must to have as active ingredients only products derived from plants. The quality control of these medicines has not yet properly established criteria even with the growing consumption of these drugs worldwide. Among the assays to evaluate the quality there is the test of dissolution, used to examine the release of the active principle from a capsule or tablet in medium that simulates the place of their absorption. Among the nature drugs best known there is Passiflora incarnata L., whose possess sedative action. The C-flavonoid glycosides vitexin and isovitexin are the main components found in aereal parts of Passiflora sp and are used as markers of the genus in the tests of quality control. This study was conducted to apply the test of dissolution to phytotherapy containing passion flower. The method for quantifying of capsules and tablets used was the spectrophotometric assay described in British Pharmacopoeia (2007) to the powder of P. incarnata. The method has been validated for capsules and then used in the tests of dissolution, with apparatus shovel, 37 C ± 0.5 ° C, speed of 50 rpm in 500 mL of the medium of dissolution 0.1 M HCl pH 1.2, collecting 50 ml at the end of 30 minutes. We have investigated an alternative method for quantification of total flavonoids in ultraviolet spectrophotometer , in a wavelength of 269 nm or 350 nm, looking for a more simple and quick method to be used in routine testing, and because be less polluting and unhealthy. In the tests carried out to quantify the levels of total flavonoids found for the manipulated capsules were 2% for powder and 0.975 % for dry extract; for industrialized tablets the levels were 5.5% and 1.52%. The dissolution assay of the pharmaceutical forms at this study showed that the capsules containing dry extract or powder and one of the industrialized tablets showed 87%, 75% e 76%, respectively, of dissolution within 30 minutes of tests. In relation to the proposed alternative method, was selected the wavelength 350 nm for direct quantification of flavonoids presents in the phytomedicines and at the medium of dissolution. It can be noted that the results were consistent and similar to the datas of the validated method: 85% to dry extract, 80% to powder e 75% to one of the industrialized tablets. Whereas the phytomedicines represent a niche market and an opportunity for diversification of the national pharmaceutical industry, it is necessary to defined parameters of quality to ensure its therapeutic action. So, the studies of dissolution are an essential tool which helps to ensuring the quality of these medicines.Item Produção e caracterização da atividade de tirosinase no extrato bruto de Pycnoporus sanguineus CCT-4518(Universidade Federal de Goiás, 2009-07-15) DUARTE, Livia Teixeira; SANTIAGO, Mariângela Fontes; http://lattes.cnpq.br/7143224488081563; BARA, Maria Teresa Freitas; http://lattes.cnpq.br/3914164125498267Tyrosinase (E.C.1.14.18.1) is an enzyme of industrial interest that catalyses the ohydroxylation of monophenols (monophenolase activity) and the oxidation of o-diphenols to reactive o-quinones (diphenolase activity), both reactions using molecular oxygen. Pycnoporus sanguineus (L. ex Fries) Murril, is a white rot fungi capable of producing tyrosinase and widely distributed in nature. It is found in regions of mild climate and in tropical forest. The production and characterization of tyrosinase from P. sanguineus were investigated. The selection of inductors, determination of the luminosity influence, biomass and culture media in the production of tyrosinase and the effect of inhibitors on enzyme activity were determined. The fungus produced intracellular tyrosinase and the higher activity was observed using 0.15% L-tyrosine as inducer, in the presence of light, with inoculum of 10 mycelium discs, medium malt extract broth 2%, incubation at 30°C, and constant agitation of 150 rpm, during 2 days. 6 mmol.L-1 salicylhydroxamic acid (SHAM) and 6 mmol.L-1 phenylthiourea (PTU) inhibited 100% of the tyrosinase activity. 0.1 mmol.L-1 sodium azide inhibited 4.15% of tyrosinase activity, while no inhibition was observed after addition of 0.1 mmol.L-1 of phenylmethanesulfonyl fluoride (PMSF). Using L-dopa as substrate, the intracellular crude extract presented optimum pH of 6,6, optimum temperature of 45°C, low stability at 50°C, maintaining about 50% of the activity after 15 min of incubation. The tyrosinase production was confirmed by non-denaturing polyacrylamide gel electrophoresis, using commercial fungal tyrosinase as positive controlItem Ensaio de dissolução de fitoterápicos contendo Maytenus ilicifolia Mart ex Reiss (Celastraceae): validação de métodos para quantificação de catequina e epicatequina(Universidade Federal de Goiás, 2010-12-17) LEAL, Ana Flavia Vasconcelos Borges; DINIZ, Danielle Guimarães Almeida; http://lattes.cnpq.br/6801755844853116; BARA, Maria Teresa Freitas; http://lattes.cnpq.br/3914164125498267Maytenus ilicifolia Mart ex Reiss (Celastraceae) popularly known in Brazil as espinheirasanta is used against gastric disorders and ulcers. Quality of herbal medicines is essential to ensure the safety and efficacy of the drug. The dissolution test is an essential tool to assess the quality of herbal medicines in the solid dosage form for oral use. This work aimed to evaluate the dissolution behavior of eleven herbal medicines containing Maytenus ilicifolia, produced with powder or dry extract, purchased in local shops (Goiânia-GO, Brazil). Assay of catechin and epicatechin by high-pressure liquid chromatography (HPLC) and dissolution methods were validated and obtained results allowed the quantification of these markers with precision, accuracy and selectivity. The dissolution profiles have been determined adopting the following conditions: paddle apparatus at 75 rpm and HCl 0.1 M (900 mL at 37°C ± 0.5°C) as dissolution medium. Through the dissolution profiles it has been determined the mathematical model of release (first order), the percentage of catechin and epicatechin dissolved at 45 minutes and the efficiency of dissolution. Capsule containing powder showed rapid release of the markers in the medium (Q>85% in 12 minutes). Tests conducted to verify the quality of herbal medicines were average weight, weight variation, dissolution, chromatographic profile and sensory analysis. It was found that only 45.5% of analyzed herbal medicines products provide satisfactory results. The dissolution test, sensory analysis, average weight, weight variation and chromatographic profile were responsible for noncompliance of, respectively, 18.2%, 18.2%, 18.2%, 9.1% and 36.4% of the samples. Obtained data demonstrated the need to implement these tests in quality control of herbal medicines, so by investigating the release of the active ingredients of these medicines.Item Obtenção e Caracterização do Extrato Seco Padronizado dos Frutos da Sucupira Pterodon emarginatus Vogel, Fabaceae(Universidade Federal de Goiás, 2010-08-31) OLIVEIRA, Patrícya Caixeta de; CONCEIÇÃO, Edemilson Cardoso da; http://lattes.cnpq.br/7193007113950510; BARA, Maria Teresa Freitas; http://lattes.cnpq.br/3914164125498267The Pterodon emarginatus Vogel, known as sucupira branca (Fabaceae) has been shown to possess larvicidal activity against (Aedes aegypti), antiparasitic effect (Leishmania amazonensis), fungicide, bactericide, antioxidant, antiinflammatory, antinociceptive, and antiproliferative activity among several different human cancer cell lines that may be related to presence of diterpenes, in particular, vouacapanes derivatives. This work aimed to define and characterize the powder, the crude and standardized dry extract of the fruit of sucupira . After crushing the fruits, showed 1.2% volatile. The chromatographic (TLC) indicated the presence of chemical markers vouacapanes (6α,7β-dihydroxyvouacapan-17β-oic acid - Rf = 0.16; 6α,7β-dihydroxyvouacapan-17β-oate methyl ester - Rf = 0.44; 6α-hydroxyvouacapan-7β-17β-lactone - Rf = 0.71) and lupeol (Rf = 0.78). In vibration bands in the infrared region coinciding with the five main bands of vouacapanes (-OH, C-H , C=O, C=C, C-O). The concentration of total terpenes of 9.3%. Then to obtain the crude extract was held percolation in 95% ethanol PA. The crude extract showed pH 5.1, relative density of 0.87, viscosity of 25.43 mPas, solid 15.2%, total terpenes of 17.1% and the same chromatographic profile and in the infrared spectrum obtained for the powder plant. Another method used for investigation of terpenes was the gas chromatography / mass spectrometry in the hexane extract. We identified the presence of vouacapanes and the lupeol. The drying process of the crude extract was used nebulization (spray drying) using different adjuvants drying, flow of 0.19 L / h, air flow 35 L / min, inlet temperature 110°C, outlet temperature 97°C, double fluid nozzle spray type 1.2 mm. The best results were obtained with the colloidal silicon dioxide (Aerosil®) to 20%, giving an appearance of fine powder. Thermal analysis performed for the marker 6α,7β-dihydroxyvouacapan-17β-oate methyl ester showed that the drying parameters were safe by preventing the thermal degradation of diterpenes. The efficiency of the drying process was 16.2%, degradation of 10.9% and the concentration of terpenes total in dry extract was 13.9%. The chromatographic profile was maintained and the volatile content was 3.5%. The photomicrographs showed irregular and spherical particles, rough surface. The analytical method by spectrophotometry in the visible region for the quantification of total terpenes in the dust, crude extract and dry extract was validated presenting selective, linear, precise, accurate and robust. In the test of antinociceptive activity (capsaicin), the pre-treatment with dry extract of sucupira (SDE 1000 mg / kg + 20% DMSO) reduced the time reactivity in 50.9% compared to the vehicle. The results may suggest that the technological processes employed to transform the fruits of sucupira in standardized dry extract was adequate to maintain quality chemistry and antinociceptive activity described for the fruits. This work represents the first description of the taking of standardized dry extract and identification of lupeol in fruits of sucupira .Item Obtenção e padronização de extrato seco de Mikania laevigata (guaco)(Universidade Federal de Goiás, 2011-05-31) PINTO, Mariana Viana; CONCEIÇÃO, Edemilson Cardoso da; http://lattes.cnpq.br/7193007113950510; BARA, Maria Teresa Freitas; http://lattes.cnpq.br/3914164125498267Mikania laevigata Sch.Bip. ex Baker., popularly known as guaco, is a Brazilian medicinal plant belonging to Asteraceae family and widely used in medicine to treat coughs and respiratory disorders. It presents bronchodilator and antiulcer activities among others. Coumarins, terpenes and organic acids are the main chemical constituents of this specie. In the literature no studies were found concerning the production of standardized dried extracts from this plant specie, therefore the aim of this study was the obtaining and the characterization of standardized dry extract of this plant specie. M. laevigata leaves were collected, dried and crushed, resulting in the plant drug. The hydroalcoholic extract was obtained by percolation of the drug plant and the extraction liquid used was ethanol 80% (v/v). After drying experiments were carried out in order to investigate the influences of the extract feed rate and the drying adjuvant ratio in the characteristics of the dried extracts obtained.-coumaric acid and coumarin were used as chemical markers and were quantified by High Performance Liquid Chromatography (HPLC) in all stages of the process. An analytical method was validated following the criteria proposed by the Agência Nacional de Vigilância Sanitária (ANVISA), proving to be selective, linear, precise, accurate, sensitive and robust. In the characterization of plant drug the volatile content obtained was 9.00 ± 1.75%, the coumarin content was 0.72 ± 0.02% and the acid -coumaric content was 0.13 ± 0.005%. The average particle size was 444.823 m and swelling index was 3.2 in ethanol 80%, 4.27 in ethanol 50% and 10.13 in water. The hydroalcoholic extract presented a density of 0.9256 ± 0.0006 g / mL, total solids content of 5.18 ± 0.21%, pH 6.1 ± 0.00, alcohol content of 56% and viscosity of 5.57 mPas. The drying yields ranged from 11.35 to 54.11%, and all dried products presented volatiles content and water activity below 5% and 0.5, respectively. The selected condition to be used to obtain standardized dried extracts of M. laevigata was that performed with a smaller proportion of colloidal silicon dioxide (15%) and increased extract feed rate (6mL/min), in which levels were obtained coumarin and -coumaric acid content of 1.83 ± 0.004% and 0.80 ± 0.012%, respectively. The dry extract obtained in these conditions showed spherical particles and their antiulcer activity in the model of indomethacin induced ulcers was demonstrated, suggesting a gastroprotective effect of this extract. Comparison of powder, hydroalcoholic and dry extract chromatographic (TLC and HPLC) show that the hydroalcoholic extraction was suitable for the extraction of coumarin and -coumaric acid and the drying process did not cause changes in the chemical profile of the extract in relation to these substances. Thus, the production of dried extracts by spray drying from the leaves of M. laevigata can be an interesting and promising alternative for obtaining such an intermediate product.Item Obtenção e caracterização do extrato seco Padronizado de Vernonanthura ferruginea (LESS.) H. Rob., Asteraceae (Assa-Peixe)(Universidade Federal de Goiás, 2011-04-18) SILVA, Ezequiane Machado da; CONCEIÇÃO, Edemilson Cardoso da; http://lattes.cnpq.br/7193007113950510; BARA, Maria Teresa Freitas; http://lattes.cnpq.br/3914164125498267Vernonanthura ferruginea (Less.) H. Rob. (Asteraceae), a typical plant of the Brazilian savannah, popularly known as assa-peixe, presented analgesic, antiedematogenic, antioxidant and antiulcer activities in scientific studies. But in literature there are no works related to the production of dried extracts of this plant species. In this work, standardized spray-dried extracts of V. ferruginea were obtained. The work started with the obtainment and characterization powder of the leaves of the V. ferruginea. A hydroalcoholic extract was obtained by percolation using as solvent ethanol 95 % (v/v). The drying experiments followed a 23 + 2 factorial design. The influence of several in-process parameters e. g. extract feed rate, drying air inlet temperature and drying adjuvant ratio, on the dried extracts properties were investigated. The properties investigated were process yield, residual moisture, water activity, total triterpenes content and lupeol content. The lupeol, triterpene related to several pharmacological activities (anti-inflammatory, anticancer, antiulcer, etc), was first time isolated in this species and was used as chemical marker in High Performance Liquid Chromatography (HPLC) quantification assays in all steps of this work. In this way an analytical methodology was validated following Agência Nacional de Vigilância Sanitária (ANVISA Brazilian National Health Surveillance Agency) guidelines. This methodology proved to be selective, linear, sensible, precise, accurate and robust. On the herbal raw material characterization, preliminary phytochemical screening was performed, which confirmed the presence of tannins, flavonoids, triterpenes, saponins, resins and mucilage. Was also observed a mean powder size of 398.97 ± 6.91 μm, as well as moisture, total triterpenes and lupeol contents of 8.25 ± 0.57%, 2.49 ± 0.16% and 0.17 ± 0.01%, respectively. The concentrated hydroalcoholic extract presented density of 0.8364 ± 0.0015 g/mL, solids content of 6.70 ± 0.19%, pH 5.81 ± 0.05, alcoholic content of 87.08 ± 0.80% and a viscosity of 4.93 ± 0.0416 mPas. The levels of total triterpenes and lupeol contents were 22.57 ± 0.94 and 4.31 ± 0.01%, respectively. Results showed drying yields ranging from 10.47 to 30.82%. Also, all dried products showed moisture contents and water activities below 5% and 0.5, respectively. Analysis of variance (ANOVAs) proved that all factors and some of their interactions significantly affected the levels of total triterpenes and lupeol contents. Moreover, the selected condition for obtaining dry extracts of V. ferruginea with adequate physicochemical properties is the one performed with the higher colloidal silicon dioxide ratio (20%), the lower extract feed rate (4mL/min) and the lower drying air inlet temperature (80°C), in which it was obtained levels of total triterpenes and lupeol contents of 7.65 ± 0.27% and 2.47 ± 0.04%, respectively. The dry extract obtained in this condition show high content of spherical particles and its antiulcer activitie in model of ulcer induced by indomethacin was proved. Comparison of powder, hydroalcoholic and dry extract chromatographic (TLC) show that the hydroalcoholic extraction was selective for the extraction of lupeol against other major terpenes and the drying process did not cause changes in the chemical profile of the extract with respect to these substances. Thus, the spray drying can be an attractive e and promising alternative for the development of V. ferruginea phytomedicines.Item DESENVOLVIMENTO E VALIDAÇÃO DE MÉTODO ANALÍTICO DE CROMATOGRAFIA LIQUIDA DE ALTA EFICIÊNCIA E ENSAIO DE DISSOLUÇÃO PARA AVALIAÇÃO DA QUALIDADE DE FITOTERÁPICOS CONTENDO Paullinia cupana Kunth(Universidade Federal de Goiás, 2009-07-30) SOUSA, Sandra Alves de; BARA, Maria Teresa Freitas; http://lattes.cnpq.br/3914164125498267The Herbal medicines are obtained by using only raw vegetable active. Guarana (Paullinia cupana Kunth.) is used as an invigoranting and challenging due to the presence in the seeds of methylxanthines, especially caffeine and theobromine and theophylline lesser amount, and tannins. The test is an dissolution of the tests used in assessing the quality of these medicines and throught this you can evalueted the release of the active principle from a solid form in an environment that simulates the location of its absorption. This study aimed to evaluete the behavior of dissolution of capsules and tablets containinf guarana. This study investigated initially, spectrophotometric and chromatographic methods for quantification of tannins and methylxanthines in pharmaceutical forms and the means of dissolution. The results obtained by spectophotometry showed lack of specificity of this method under our experimental conditions for quantification of methylxanthines. In the analysis of tannins, the procedure is not feasible for an routine laboratory to be long and larorious. The method by High Performance Liquid Chromatography allowed the quantification of the five markers simultaneously, with precision, accuracy, selectivity and robustness. The dissolution test using apparatus as man, 37 ºC ± 0,5 ºC, speed of 75 rpm in 900 mL of 0.1 M HCl medium pH 1,2 provided the best results was validated and used in assessing the behaviour of dissolution of the capsules and tablets based on guarana. It was found that all markers were quantified by the method are dissolved between 75% and 80% in time of 10 minutes. An analysis of the capsules did not break in any of the conditions employed. For the tablet was also observed in non-compliance, non-identification of catechin and epicatechin in the finished product, worrying factor, as the phenolic compounds are markers that should be present in the guarana powder. Considering the increasing use of plants is a clear need for definition of quality standards to ensure its therapeutic action and accordingly the study of dissolution is an essential tool in ensuring the quality of plants