FundaÃÃo de Amparo à Pesquisa do Estado do Cearà / Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Foram desenvolvidos e validados trÃs mÃtodos robustos para a determinaÃÃo de amoxicilina, norfloxacino, oxcarbazepina (OXC) e 10,11-dihidro-10-hidroxicarbamazepina (MHD) em plasma, utilizando cromatografia lÃquida de alta eficiÃncia em fase reversa (RP-HPLC) com detecÃÃo ultravioleta, fluorescÃncia e espectrometria de massa, respectivamente. Os mÃtodos envolveram extraÃÃo lÃquido-lÃquido, com diclorometano (amoxicilina), acetonitrila (amoxicilina e norfloxacino), Ãter etÃlico-diclorometano (60:40 v/v, oxcarbazepina), utilizando cefadroxil, ciprofloxacino e d10-carbamazepina como padrÃes interno (PI). SeparaÃÃes cromatogrÃficas foram realizadas utilizando colunas Gemini C18 5 Âm (150 X 4,6 mm), Synergi RP-MAX 4 Âm (150 X 4,6 mm) e Luna C18 Âm (150 X 4,6 mm), com sistemas de eluiÃÃo constituÃdos por mistura de tampÃo fosfato de 0,01 M (pH 3,5)/acetonitrila (95:5 v/v), acetonitrila-tampÃo fosfato (85:15, v/v) e acetonitrila-Ãgua (50:50 v/v) + 20 mM Ãcido acÃtico, respectivamente. As curvas de calibraÃÃo foram lineares, nas faixas de 0,5 a 40 Âg/mL, 30 a 3500 ng/mL, 20 a 5250 ng/ml e 40 a 10500 ng/ml. As recuperaÃÃes nas concentraÃÃes de 1,5, 15 e 30 Âg/ml foram de 59,4%, 60,5% e 67,1% para amoxicilina; 90, 1400 e 2800 ng/mL foram de 103,5%, 100,2% e 100,2% para norfloxacino, 60, 2000 e 4000 ng/ml foram de 105,4, 89,2 e 92,8% para OXC e 120, 4000 e 8000 ng/ml foram de 88,4, 88,7 e 90,6% para MHD, respectivamente. Os mÃtodos validados incluÃram avaliaÃÃo de precisÃo e exatidÃo intra e interlote, assegurando que estes estavam dentro de limites admissÃveis. Os mÃtodos foram entÃo aplicados com sucesso em estudos de bioequivalÃncia, administrando 500 mg ou 400 mg das formulaÃÃes referÃncia/teste de amoxicilina e norfloxacino, e em estudos farmacocinÃticos com formulaÃÃo de oxcarbazepina suspensÃo (6%), em voluntÃrios sadios. / A robust method for the determination of amoxicillin, norfloxacin, oxcarbazepine (OXC) and its active metabolite, 10,11-dihydro-10-hydroxycarbamazepine (MHD) in human plasma, using reversed-phase high-performance liquid chromatography (RP-HPLC) with ultraviolet, fluorescence and mass spectrometry detection, respectively, have been developed and valited. The methods involve precipitation of plasma protein with dichloromethane (amoxicillin), acetonitrile (amoxicillin and norfloxacin) and diethyl etherâdiclhoromethane (60:40 v/v, oxcarbazepine), using cefadroxil, ciprofloxacin and deuterade carbamazepine (d10-carbamazepine) as internal standard (IS). Chromatographic separations were performed on a column Gemini C18 5 Âm (150 X 4.6 mm), Synergi MAX-RP 4 Âm (150 X 4.6 mm) and Luna C18 5 Âm (150mm X 4.6 mm) with an elution system consisting of a mixture of 0.01 M buffer phosphate (pH 3.5)/acetonitrile (95:05 v/v), phosphate bufferâacetonitrile (85:15, v/v) and acetonitrile/water (50:50 v/v) + 20mM acetic acid, respectively. The calibration curve was linear, in the range of 0.5 to 40 Âg/mL, 30 to 3500 ng/mL, 20 to 5250 ng/mL and 40 to 10,500 ng/mL. The recoveries at concentrations of 1.5, 15 and 30 Âg/mL were foram 59.4%, 60.5% and 67.1% for amoxicillin; 90, 1400 and 2800 ng/mL were 103.5%, 100.2% and 100.2% for norfloxacin, 60, 2000 and 4000 ng/mL were 105.4, 89.2 and 92.8% for OXC and 120, 4000 and 8000 ng/mL were 88.4, 88.7 and 90.6% for MHD, respectively. The statistical evaluation of the developed method was conducted by examining within-batch and between-batch precision data, which were within the required limits. The methods were successfully applied in bioequivalence studies given 500 or 400-mg of the reference formulation / test amoxicillin and norfloxacin, and pharmacokinetic studies with formulation of oxcarbazepine suspension (6%) in healthy volunteers.

Magister Pharmaceuticae - MPharm / Many active pharmaceutical ingredients (APIs) have poor physicochemical properties such as solubility, dissolution and chemical stability. Several strategies are used to enhance and improve these properties of the API. Co-crystallization and polymorphism studies are possible strategy used in pre-formulation studies to optimize these properties of the drug without modifying its pharmacological effect. The purpose of this research was to investigate the polymorphism and co-crystallization effects of the penicillin-type antibiotic, amoxicillin tri-hydrate. Several techniques such as: slow evaporation, slow cooling, vapour diffusion, sublimation, grinding and solvent assisted grinding was employed. In producing co-crystals, the API was non-covalently bound to selected co-formers such as: saccharin, nicotinamide, salicylic acid, L-tartaric acid, D-tartaric acid, L-aspartic acid, stearic acid, benzoic anhydride, oxalic acid di-hydrate, cinnamic acid, succinic acid and citric acid monohydrate. Nine co-crystals of amoxicillin tri-hydrate had been formed. Differential scanning calorimetry (DSC), hot stage microscopy (HSM) and thermal gravimetric analysis (TGA) was conducted to analyse the thermal behaviour of the co-crystals. Powder X-ray diffraction (PXRD) and spectroscopic techniques [infra-red (FTIR) and H1-nuclear magnetic resonance (H1MNR)] were employed for screening of the co-crystal forms. Furthermore, dissolution testing was conducted to investigate the application of the newly derivatised forms.

The existence of pharmaceuticals in the environment has some adverse effects, and may pose threat to the organisms in the environment. The possibility of removing certain pharmaceuticals from wastewater was tested using Octolig®, a commercially available material with polyethyldiamine moieties covalently attached to high-surface area silica gel. Selected drug models were subjected to column chromatography in efforts to effect removal by means of ion encapsulation, the effectiveness of which would depend upon having appropriate anionic functional groups. The experimental results suggested that the model compounds, Rose Bengal, Eosin Y, Erythrosine , ZPS, and Lissamine Green B were successfully encapsulated by Octolig®, while Methylene Blue with quaternary ammonium groups was (statistically) not. In contrast, complete success was attained for removing of each of three xanthenylbenzenes (Rose Bengal, Eosin Y, Erythrosine) that have both phenolic and carboxylic acid groups. In addition complete success was attained for ZPS (zinc phthalocyaninetetrasulfonate) with sulfonate groups present. A test of a real pharmaceutical compound, Amoxicillin, indicated that Octolig® can be used to remove this compound from aqueous media.

The efficacy of short term (3 day) oral amoxicillin therapy was compared to conventional (14 day) oral therapy in an experimental model of bacterial lower urinary tract infection (UTI) in the cat. Chemical cystitis was induced using an infusion solution of salicylic acid, 70% ethanol, and normal saline via transabdominal cystocentesis. Cats were challenged with a Staphylococcus intermedius inoculum twenty-four hours later introduced via urethral catheterization. Serial quantitative aerobic bacterial urine cultures obtained via cystocentesis were used to evaluate groups of cats. Eighteen adult cats (9 males and 9 females) were divided into 3 groups of 6 cats (3 males and 3 females): Group I = conventional amoxicillin therapy (14 day), Group II = control group (no treatment), and Group III = short term therapy (3 day). Results indicated the conventional therapy successfully eradicated infection, however, the short term therapy did not eradicate infection when compared to controls. During the study period the diuretic furosemide was used in some cats to facilitate cystocentesis procedures. Those cats were observed to exhibit less stranguria, which is a common sign of lower UTI. The second study evolved from observations made in the first study and evaluated the effect of furosemide on conventional antibiotic therapy in an experimental model of bacterial lower UTI in the cat. A similar experimental design was utilized with Group I = control group (no treatment), Group II = oral furosemide (14 day), and Group III = oral furosemide and oral amoxicillin (14 day). Statistical analysis failed to demonstrate the efficacy of the furosemide and amoxicillin combination, but showed furosemide alone was not an appropriate therapy when compared to controls. It was again observed that those cats receiving furosemide showed fewer secondary signs of lower UTI such as stranguria which suggests a possible role for furosemide as adjunct therapy in the treatment of lower UTI in the cat. / Master of Science

Die Kontamination von Trinkwasser mit Arzneimitteln stellt eine ernste Gesundheitsgefahr dar. Um die Trinkwasserqualität kontinuierlich überwachen und im Falle einer Verunreinigung zeitnah reagieren zu können, sind neuartige Sensoren erforderlich. Hier können immunanalytische Methoden, die auf der Bindung des Analyten an hochselektive Antikörper beruhen, hilfreich sein. In dieser Arbeit wurden magnetpartikelbasierte Immunoassays (MBBAs) für zwei relevante Kontaminanten des Trinkwassers entwickelt: Diclofenac (DCF) und Amoxicillin (AMX). Bei letzterem erwiesen sich neben der Ausgangsverbindung auch dessen Hydrolyseprodukte (HPs) als relevant für die Gefährdungsbeurteilung. In einer umfassenden Studie wurde der Einfluss von externen Faktoren und intrinsischen Eigenschaften des Wassers auf die Hydrolysegeschwindigkeit untersucht. Da die Hydrolyse von AMX auch die Erkennung durch den Antikörper beeinflusst, wurde eine Strategie zur Analyse von Proben mit unbekanntem Hydrolysegrad von AMX unter Verwendung des Enzyms β-Lactamase in der Probenvorbereitung entwickelt. Für beide Analyten ermöglichen die MBBAs eine schnelle Quantifizierung mit Ergebnissen in weniger als einer Stunde, was eine wesentliche Verbesserung gegenüber herkömmlichen Immunoassays wie dem Enzyme-linked Immunosorbent Assay (ELISA) darstellt. Im Vergleich zu den entsprechenden ELISAs mit denselben Antikörpern weisen die MBBAs zudem verbesserte analytische Parameter auf, wie einen breiteren Messbereich und niedrigere Nachweisgrenzen. Aufgrund der magnetischen Eigenschaften der Partikel, die als Plattform für die Assays dienen, eignen sie sich für den mobilen und automatisierten Einsatz vor Ort. Ein integriertes Diagnosesystem, bei dem die elektrochemische Detektion mittels Chronoamperometrie auf einem mikrofluidischen Chip eine weitere Miniaturisierung des Systems ermöglicht, wurde entworfen, um die Überwachung der Trinkwasserqualität online in Wasserwerken zu ermöglichen. / The contamination of drinking water with pharmaceuticals represents a severe health risk. In order to monitor the drinking water quality continuously and enable quick countermeasures in case of contamination, novel sensors are required. Here, immunoanalytical methods based on the binding of the analyte to highly selective antibodies can be helpful. In this work, magnetic bead-based immunoassays (MBBAs) have been developed for the detection of two relevant contaminants of drinking water: diclofenac (DCF) and amoxicillin (AMX). In case of the latter, not only the parent drug is of interest in the risk assessment but also its hydrolysis products (HPs). In a comprehensive study, the influence of external factors and intrinsic properties of the water on the rate of hydrolysis was investigated. As the hydrolysis of AMX further impacts the recognition by the antibody, a strategy to analyze samples with unknown hydrolysis degree of AMX was established employing the enzyme β-lactamase in sample preparation. For both analytes, the MBBAs enable the fast quantification with results obtained in less than one hour which represents a major improvement over conventional immunoassays like the enzyme-linked immunosorbent assay (ELISA). Compared to the respective ELISAs with the same antibodies, the MBBAs further exhibit improved analytical parameters such as a broader measurement range and lower limits of detection. Due to the magnetic properties of the beads that serve as a platform for the assays, they are suitable for the mobile and automated detection at the point-of-care. An integrated diagnostic system was designed in which electrochemical detection with chronoamperometry on a microfluidic chip allows for further miniaturization of the system to enable monitoring of the drinking water quality online in water supply pipes at waterworks.

O presente trabalho estuda a remoção dos fármacos amoxicilina (AMX), diclofenaco sódico (DCF) e paracetamol (PAR) em solução aquosa pelos processos de adsorção em batelada e coluna de leito fixo utilizando o carvão ativado granulado (CAG) como adsorvente. Os experimentos foram realizados para cada fármaco de forma independente. Na adsorção em batelada foram avaliadas as influências do pH (2 – 10), concentração de adsorvente (5 – 20 g L-1) e tempo de contato (5 – 350 min). Foi realizada a investigação da cinética de adsorção e também do equilíbrio de adsorção através de isotermas nas temperaturas de 25, 35 e 45 ºC. A adsorção em leito fixo foi estudada através de planejamentos experimentais, onde foram avaliados os efeitos da concentração inicial do poluente (C0: 20 – 100 mg L-1), massa do leito (W: 0,5 – 1,5 g) e vazão (Q: 3 – 5 mL min-1) sobre o tempo de saturação (tsat) e a quantidade adsorvida (qsat). Os modelos de Thomas, Bohart-Adams e Yan, além de um modelo desenvolvido no software EMSO foram utilizados para análise das curvas de ruptura. O CAG utilizado apresentou área BET de 463 m² g-1 e maior volume de microporos, de 0,20 cm³ g-1. Os experimentos em batelada mostraram que o pH não teve influência significativa sobre a remoção dos três fármacos. O equilíbrio de adsorção da AMX e do DCF foi atingido após 150 min e do PAR após 180 min. O modelo de PSO foi o que melhor representou a cinética de adsorção dos três fármacos. A isoterma de Langmuir descreveu o equilíbrio da AMX a 25 e 35 ºC, e o modelo de Sips a 45 ºC. Já a adsorção do DCF foi representada pela isoterma de Freundlich e o PAR pela de Redlich-Peterson. O estudo termodinâmico indicou que a adsorção dos três fármacos foi espontânea e favorável, além de aumentar com o aumento da temperatura. Na adsorção em leito fixo, foi observado menores valores de tsat com o aumento de C0 e de Q e diminuição de W. Foi verificado que qsat aumentou com o aumento de C0 e diminuição da Q para o planejamento do PAR, onde essa variável foi significativa. Já o aumento de W aumentou qsat no planejamento do PAR e diminuiu nos casos da AMX e DCF. A AMX foi o poluente que apresentou os menores tempos de saturação, seguido do DCF e do PAR, na adsorção em leito fixo. Foi constatado que o modelo de Yan foi o que melhor reproduziu o comportamento das curvas de ruptura para os três fármacos, na comparação com os outros modelos analíticos e com o modelo numérico proposto no software EMSO. De forma geral, foi verificado que os processos de adsorção tanto em batelada quanto em leito fixo apresentam potencial de aplicação como alternativa de tratamento avançado de água e efluentes que contenham fármacos. / The present work studies the removal of amoxicillin (AMX), sodium diclofenac (DCF), and paracetamol (PAR) from water by adsorption onto granular activated carbon in batch process and fixed bed column. Batch adsorption experiments were performed to evaluate the influence of pH (2 – 10), adsorbent concentration (5 – 20 g L-1) and contact time (5 – 350 min). Pseudo-first order, pseudo-second order and intraparticle diffusion models were evaluated in the kinetics investigation. Equilibrium adsorption was investigated using Langmuir, Freundlich, Sips and Redlich-Peterson equations. Fixed bed adsorption was studied through experimental design to evaluate initial contaminant concentration (C0, 20 – 100 mg L-1), amount of adsorbent (W, 0.5 – 1.5 g) and feed flow rate (Q, 3 – 5 mL min-1) effects. The analytical models of Thomas, Bohart-Adams and Yan were selected to investigate the breakthrough curves behavior. In addition, a numerical model was developed and solved using EMSO software. The granular activated carbon (GAC) used had BET surface area of 463 m² g-1 and volume of 0.20 cm³ g-1 of micropores. The pH had no significant effect on the adsorption removal of the three drugs. Adsorption equilibrium of AMX and DCF was reached after 150 min and 180 min for PAR. Pseudo second order model best represented kinetic adsorption of the three compounds. At best conditions in batch process, adsorbent concentration was 12.5 g L-1 for AMX and DCF and 10 g L-1 for PAR. Langmuir isotherm best described AMX adsorption equilibrium at 25 and 35 ºC, and Sips model at 45 ºC. DCF and PAR adsorption followed the Freundlich isotherm and Redlich-Peterson model, respectively. Thermodynamic study indicated that the three drugs adsorption were spontaneous and favorable processes. In addition, adsorption increased at higher temperatures. In fixed bed adsorption experiments, saturation time (tsat) decreased with the increase of initial concentration and flow rate for both drugs. W had positive effect on tsat. The amount adsorbed (qsat) was enhanced at higher C0 and lower Q. qsat was higher at higher Q for AMX and DCF and lower Q for PAR adsorption. Yan model best reproduced breakthrough curves behavior for all drugs among the analytical models and the numerical model developed on EMSO software. Thus, adsorption processes in batch mode and fixed bed column showed to be effective for the removal of drugs of different therapeutic classes from water.

O uso de antibióticos para reduzir a infecção pós-operatória em cirurgia de terceiro molar permanece controverso. O objetivo deste estudo foi avaliar a eficácia da terapêutica com Amoxicilina em múltiplas doses para a prevenção de infecção em pacientes submetidos à extração de terceiro molar inferior com posição 2B (classificação de Pell & Gregory). O estudo foi prospectivo, cego, randomizado, duplo, controlado por placebo, com 32 pacientes. Cada paciente atuou como seu próprio controle usando a técnica de boca dividida. Um terceiro molar inferior foi retirado sob a cobertura de antibióticos no pós-operatório (500 mg de amoxicilina a cada 8 horas durante 7 dias) e o outro foi removido sem cobertura antibiótica pós-operatória (cápsulas de placebo a cada 8 horas durante 7 dias), mas em ambas cirurgias foi administrada dose única no pré-operatório de 1 g de amoxicilina. Os seguintes parâmetros foram avaliados no pré-operatório, no quarto e sétimo dias pós-operatórios: trismo, edema facial, temperatura corporal, linfadenopatia, infecção, disfagia e dor. Não houve diferença estatisticamente significativa nos parâmetros avaliados entre os pacientes operados (p> 0,05). Resultado do estudo mostrou que a administração de amoxicilina em dose única pré-operatória e em doses múltiplas pós-operatórias não se mostrou mais eficaz do que a administração somente em dose única pré-operatória com relação aos parâmetros clínicos avaliados nas exodontias de terceiros molares inferiores. / The use of antibiotics to reduce postoperative infection in third molar surgery remains controversial. The goal of this study was to evaluate the efficacy of multi-dose Amoxicilina therapy for the prevention of infection in patients undergoing lower third molar extraction with position 2B (Pell & Gregory classification). The study was a prospective, randomized, double blind, placebo-controlled trial with 32 patients. Each patient acted as their own control using the split-mouth technique. One lower third molar were removed under antibiotic cover postoperative (500 mg amoxicillin 8 hourly for 7 days) and the other were removed without antibiotic cover postoperative (placebo capsules 8 hourly for 7 days), but both surgery was administered preoperatively single dose of 1g of amoxicillin. The following parameters were evaluated on the preoperative and fourth, seventh days postoperative: trismus, facial swelling, body temperature, Lymphadenopathy, infection, dysphagia and pain. There was no statistically significant difference in the parameters evaluated between patients operated (p > 0.05). Results of the study showed that administration of amoxicillin single dose preoperative and postoperative multiple doses was not more effective than single dose administration only pre-operative with respect to clinical parameters evaluated in the lower third molar extractions.

Orientador: Paulo Câmara Marques Pereira / Resumo: O trato gastrointestinal possui enorme variedade de bactérias aeróbias e anaeróbias, metabolicamente ativas, que interagem entre si em complexo ecossistema. Essa microbiota desempenha funções importantes no metabolismo, nutrição, imunidade e proteção contra a colonização por microrganismos patogênicos. Vários fatores podem influenciar essa microbiota, entre eles a idade, dieta, processos inflamatórios e infecciosos e utilização de antimicrobianos. Assim, este estudo foi desenvolvido com o objetivo de verificar a influência das infecções bacterianas do trato respiratório e do tratamento com amoxicilina sobre a microbiota intestinal normal do paciente. O processo infeccioso bacteriano do trato respiratório influenciou a microbiota intestinal dos pacientes. Houve redução significativa na quantidade de UFC/g de fezes de microrganismos dos gêneros Bacteroides e Lactobacillus. A utilização de amoxicilina também influenciou a microbiota intestinal. Foi observada redução significativa na quantidade de UFC/g de fezes de microrganismos dos gêneros Bifidobacterium e Lactobacillus. É importante identificar modificações na composição da microbiota intestinal, uma vez que a diminuição destes microrganismos pode representar diversos prejuízos para o hospedeiro. Um destes prejuízos é a diminuição da resistência à colonização. Esses prejuízos, quando acompanhados, podem ser minimizados pela equipe de saúde por meio de medidas relativas à conduta terapêutica e alimentar, visando reduzir a influência nociva sobre o ecossistema gastrointestinal. / Abstract: The gastrointestinal tract contains an enormous variety of metabolically active aerobic and anaerobic bacteria, that interact with each other in a complex ecosystem. That microbiota carries out important functions in the metabolism, nutrition, immunity and protection against the colonization by pathogenic microorganisms. Several factors can influence this microbiota, among them are age, diet, inflammatory and infectious processes and use of antimicrobial agents. Thus, this study was developed with the objective of verifying the influence of the bacterial infections of the respiratory tract and of the treatment using amoxicillin on the patient's normal intestinal microbiota. The bacterial infectious process of the respiratory tract influenced the patients' intestinal microbiota. There was significant reduction in the amount of CFU/g of feces of microorganisms of the Bacteroides and Lactobacillus genus. The use of amoxicillin also influenced the intestinal microbiota. Significant reduction was observed in the amount of CFU/g of feces of microorganisms of the Bifidobacterium and Lactobacillus genus. It is important to identify modifications in the composition of the intestinal microbiota, once the decrease of these microorganisms can cause damage to the host. One of these damages is the decrease in the resistance to the colonization. Those damages, when accompanied, can be minimized by the health staff using measures related to the therapeutic and alimentary conduct, seeking to reduce the noxious influence on the gastrointestinal ecosystem. / Doutor

Helicobacter pylori (hereinafter referred to as H. pylori) is a gram-negative bacteria which colonises the human stomach mucosa. Its role in the aethiopathogenesis of chronic gastritis, ulcer disorders of the gastroduodenum and MALT-lymphoma has been clearly demonstrated, and in connection with the occurrence of stomach cancer it has been indicated by the World Health Organisation (WHO) as a class I carcinogen. H. pylori infection can be detected from samples of stomach mucosa taken in an endoscopic examination (rapid urease test, microscopic examination, culture), or the non-invasive method can be used (13 C-Urea Breath Test or H. Pylori stool antigen test - HpSA). Effective therapy of H. pylori infection resides in the administration of a combination of antibiotics and a proton pump inhibitor. In recent years the resistance of bacterial strains to used antibiotics has been increasing on a worldwide scale, and we can also observe this trend in the case of H. pylori. If the level of resistance exceeds 20 % for clarithromycin and 40 % for metronidazole, these antibiotics are not recommended for the treatment of an infection caused by this bacteria. In a group of 61 patients at the Department of Internal Medicine at the University Hospital Motol who had undergone an endoscopic examination of the...

<p><font face="Tahoma"> <p align="left">The purpose of this study is to evaluate the need for prophylactic antibiotic treatment in third molar surgery and to establish specific guidelines for antibiotic prophylaxis in the department of Maxillo-Facial and Oral Surgery (MFOS) at Tygerberg Academic, Groote Schuur and Mitchells Plain Hospitals.</p> </font></p>