Mestrado em Controlo de Qualidade / MSc in Quality Control

Mestrado em Controlo de Qualidade / MSc in Quality Control / A 3,4-metilenodioximetanfetamina (MDMA; ecstasy ), tal como outras anfetaminas, tem sido considerada por muitos como sendo uma droga segura . No entanto estão descritas na literatura muitas respostas de toxicidade, reacções adversas e mortes relacionadas com a sua ingestão recreativa. Um dos seus efeitos adversos, potencialmente fatal, é a hiponatrémia. Este efeito foi relacionado com alterações na secreção da hormona antidiurética (ADH, AVP ou arginina-vasopressina) desencadeadas pela MDMA. A hiponatrémia foi apontada como causa possível para numerosas intoxicações severas e por vezes fatais decorrentes da ingestão desta droga. Estudos recentes in vivo, em humanos saudáveis do sexo masculino, e in vitro, em hipotálamo isolado de rato, demonstraram que a bioactivação metabólica da MDMA, nomeadamente a desmetilenação seguida pela O-metilação do catecol resultante, é crucial para a libertação da AVP quer in vivo quer in vitro. Para a avaliação da contribuição desta via metabólica para a expressão in vivo do efeito de hiponatrémia causado pela ingestão da MDMA, é crucial quantificar estes metabolitos e relacionar o perfil metabólico com a magnitude do efeito hiponatrémico. Com este objectivo, foi desenvolvido e validado um método de GC-MS/MS para a quantificação da MDMA e dos seus principais metabolitos, metilenodioxianfetamina (MDA), 4-hidroxi-3-metoxianfetamina (HMA) e 4-hidroxi-3-metoximetanfetamina (HMMA), no plasma e na urina. Para melhor compreender a influência da MDMA e da sua bioactivação na secreção da AVP foram realizados estudos in vivo em ratos Wistar machos e fêmeas, aos quais foi administrada MDMA na dose 20 mg/kg. Nos estudos realizados 1 hora após a administração de MDMA foram avaliados os níveis plasmáticos de AVP e as concentrações plasmáticas da MDMA e dos metabolitos MDA, HMA e HMMA. Com estas quantificações foi possível observar, nos ratos de ambos os géneros, o aumento estatisticamente significativo dos níveis de AVP em relação aos animais controlo, ao mesmo tempo que não se detectaram correlações estatisticamente significativas entre os níveis de AVP a MDMA e os metabolitos MDA, HMA e HMMA. Nos estudos realizados 24 horas após a administração de MDMA foram avaliados os níveis plasmáticos e urinários de AVP e as concentrações urinárias de MDMA, MDA, HMA e HMMA. Os resultados destas determinações demonstraram que apesar de não se detectarem diferenças significativas nas concentrações plasmáticas de AVP entre os animais tratados e os animais controlo, existem diferenças estatisticamente significativas para as concentrações urinárias de AVP, verificando-se que os animais tratados com MDMA apresentam concentrações urinárias de AVP superiores. Além disso verificou-se também que os animais tratados excretaram menos urina relativamente à água ingerida, mostrando o efeito anti-diurético desencadeado pela AVP. Neste estudo foram também estabelecidas correlações positivas e estatisticamente significativas entre os níveis de AVP e as concentrações de MDMA, MDA, HMA e HMMA. A correlação com maior significado estatístico foi estabelecida com o metabolito HMMA. Estes resultados permitiram pela primeira vez demonstrar a secreção da AVP em ratos após a administração da MDMA. Com estes estudos foi possível observar, in vivo, não só as alterações da secreção da AVP induzidas pela MDMA mas também as consequências dessas alterações nomeadamente na resposta antidiurética e o envolvimento desta resposta no efeito de hiponatrémia. Finalmente foi possível observar a contribuição da bioactivação metabólica para a secreção de AVP. Estes resultados permitem assim compreender melhor o envolvimento da MDMA e do seu metabolismo na resposta hiponatrémica. / Although considered as safe drugs by many, exaggerated responses and deaths have been reported due to 3,4-methylenedioxymethamphetamine (MDMA; ecstasy) abuse. One of the adverse effects associated with ecstasy intoxications is hyponatremia that has been related with a disruption on the release of the antidiuretic hormone (ADH or arginine-vasopressin) and pointed out as the possible cause of numerous severe and fatal intoxications after intake of this drug. Recent in vivo studies with human healthy volunteers and also in vitro studies performed with rat isolated hypothalamus have shown that the metabolic bioactivation of MDMA, namely its demethylenation followed by O-methylation of the resulting cathecol metabolite are crucial for the release of ADH both in vivo and in vitro. For the evaluation of the contribution of this metabolic pathway to the in vivo expression of the hyponatremic effect of MDMA it is crucial to quantify these metabolites, and to relate the metabolic profile with the magnitude of the hyponatremic effect. For this purpose, a GC-MS/MS method was developed to quantify MDMA and its main metabolites: methylenedioxyamphetamine (MDA), 4-hydroxy-3- methoxyamphetamine (HMA) and 4-hydroxy-3-methoxymethamphetamine (HMMA), in plasma and urine. To better understand the influence of MDMA and its metabolic bioactivation in the secretion of AVP in vivo studies were performed with male and female Wistar rats, the MDMA dose tested was 20 mg/kg. In the studies preformed 1 hour after the MDMA administration the plasmatic levels of AVP and the plasmatic concentrations of MDMA, MDA, HMA and HMMA were evaluated. The plasmatic concentrations of AVP obtained with the treated animals were compared with the concentrations obtained with the controls showing a statistically significant increase of AVP levels in the animals treated with MDMA. Correlations between the MDMA, MDA, HMA and HMMA and the AVP plasmatic levels were also preformed. No significant correletions were obtained. In the studies preformed 24 hours after the administration of MDMA the urinary and plasmatic levels of AVP were evaluated. The concentration of MDMA, MDA, HMA and HMMA were determined in plasma and urine. It was also established the ratio between the volume of ingested water and the volume of excreted urine. The plasmatic and urinary AVP concentrations obtained in the treated animals were compared with the concentrations obtained from the controls. This compairison showed significant increases of the urinary AVP levels in the treated animals. The evaluation of the correlations between the urinary concentrations of AVP and the urinary concentrations of MDMA, MDA, HMA and HMMA showed significant correlations between AVP and MDMA, MDA, HMA and HMMA. The evaluation of the ratio between the volume of ingested water and the volume of excreted urine showed that the treated animals excreted less urine in comparison with the ingested water. The studies performed with urines collected 24 hours after MDMA administration have shown significant positive correlations between AVP and the concentrations of MDMA, MDA, HMA and HMMA. The strongest correlation was established between the concentrations of HMMA and AVP. With this study it was possible to confirm the in vivo changes in the AVP secretion profile and relate those changes with the levels of MDMA, MDA, HMA and HMMA. It was also shown for the first time the induction of the secretion of AVP in male and female rats, one hour after the administration of MDMA. The consequent antidiuretic effect can be related with the hiponatremic effect.

Mestrado em Controlo de Qualidade / MSc in Quality Control

Mestrado em Controlo de Qualidade / MSc in Quality Control / Aiming for new compounds with antitumor activity, the synthesis of prenylated xanthones and prenylated and geranylated flavones was recently achieved on CEQOFFUP. In this work the potential antitumor activity of these compounds in three tumor cell lines, namely MCF-7 (human breast cancer cells expressing estrogen receptors (ER+)), MDA-MB-231 (human breast cancer cells non expressing estrogen receptors (ER-)) and NCI-H460 (non-small cell lung cancer) was evaluated. Structure-activity relationships were established highlighting the influence of prenylation and geranylation. Concerning xanthones, prenylation of 3,4-dihydroxyxanthone (XXIX) furnished more potent and selective derivates for MCF-7 (ER+) cells than the original oxygenated xanthone. Xanthone derivate XP13 showed the strongest inhibitory effect on the growth of breast adenocarcinoma cell line ER+, MCF-7 (GI50 = 5,3 M). Thus, potential estrogenic/antiestrogenic properties were investigated for this compound. No proliferative effect at low concentrations was observed for XP13 in experiments performed in steroid-free medium (RPMI-SFM). However, when high concentrations of XP13 were used, this prenylated xanthone inhibit cancer cell growth in a dose-dependent manner, being more active on MCF-7 (ER+) cell line than on MDA-MB-231 (ER -) cell line. This antiproliferative effect was not influenced by the culture medium (steroid (RPMI) or steroid-free medium (RPMI-SFM)). From these results it can be inferred that XP13 does not directly act on the estrogen receptor, suggesting that it could interfere with other signaling pathways. Moreover, XP13 enhanced the growth inhibitory action of 4-hydroxytamoxifen (4-OHT, XIV), a partial antiestrogen in estrogen sensitive breast cancer cells. Concerning flavone derivates, none of the six flavones investigated, that were resulted from prenylation and geranylation of baicalein (BAIC, XIX), presented a higher cytotoxic effect on all tumor cellular lines (MCF-7 (ER+), MDA-MB-231 (ER -) and NCI-H460) when compared to the original oxygenated flavone BAIC (XIX). However, monoprenylation in C(7) conduced to a flavone (FP2) with a selective inhibitory effect on the growth of MCF-7 (ER+) cells. Possible estrogenic/antiestrogenic properties were investigated for this compound. It was verified that in steroid-free medium (RPMI-SFM) experiments, FP2 presented a biphasic effect in vitro growth on the ER-positive MCF-7 cell line. Although at low concentrations this flavone has stimulated cell growth, at high concentrations a cell growth inhibition was observed. Then, the effect of FP2 in combination with E2 was examinated. Results showed that FP2 suppressed at low concentrations, the mitogenic effect enhanced by estrogenic stimulation, suggesting a competition for ERs. Also, the FP2 cancer cell growth inhibitory effect on MCF-7 (ER+) cells was stronger when assed in steroid free medium, i.e., in the absence of estrogenic stimulation. These results suggest a direct involvement of estrogen receptor in the proliferative/antiproliferative effect of flavone FP2. Moreover, FP2 enhanced the growth inhibitory action of partial (4-OHT, XIV) and pure (ICI 182,780, XII) antiestrogens in estrogen sensitive breast cancer cells. These results were consistent with previous reports of prenylated flavones and disclose for prenylated xanthones effects compatible with antiestrogenic activity. Thus, the present work represents a promising contribution for the prevention and treatment of hormone-dependent breast cancer.

a / Tese de mestrado integrado. Engenharia Química. Faculdade de Engenharia. Universidade do Porto. 2008

La documentación es fundamental para el cumplimiento de las Buenas Prácticas de Almacenamiento. Tiene por objetivo especificar los procedimientos de cada etapa del Sistema de Almacenamiento y los Registros de su ejecución, así como las funciones del personal involucrado. La presente monografía, establece los procedimientos a ser observados para que los productos farmacéuticos no sufran alteraciones durante su almacenamiento, asegurando que los productos sean conservados de tal forma que se disminuyan al máximo los factores que pudieran incidir sobre la calidad de los productos farmacéuticos, preservando la calidad de los mismos. Esta monografía tendrá una vigencia de un año y se someterá a una minuciosa revisión semestral, para mantenerlo actualizado, así mismo el profesional Químico Farmacéutico Responsable lo revisará regularmente para verificar la efectividad del mismo. El personal que labora en la Farmacia Universitaria de la Facultad de Farmacia y Bioquímica de la Universidad Nacional Mayor de San Marcos deberá poseer los conocimientos técnicos y la experiencia práctica para llevar a cabo la tarea que le corresponde. Así mismo deberá conocer en forma detallada la presente monografía. / -- Documentation is essential for the fulfillment of the Good Storage Practices. Its main objective is to specify the procedures of each phase in the storage system and the records of its performance as well as the functions of the people involved. The present work establishes the procedures to be notice so the pharmaceutical products do not suffer alterations during their storage, as a guarantee that the products are conserved so the factors that can influence in their quality will be reduce at maximum, preserving their quality. It will have a lifetime of 1 year and it will be under a strict examination every 6 months to keep it with the appropriate changes and also the pharmacist responsible will supervise it regularly to verify its effectiveness. People that work in the Universitaria drugstore of the Faculty of Pharmacy and Biochemistry from the San Marcos University should have all the technical knowledges and the practical experience to accomplish their job. Therefore, they should also know the present work. / Tesis

Se desarrolló una técnica analítica por cromatografía liquida de alta performance (HPLC) para cuantificar los principios activos clonixinato de lisina y pargeverina clorhidrato en tabletas recubiertas, debido a que la técnica de análisis para este medicamento compuesto no se encuentra en libros oficiales. La técnica de análisis para ambos principios activos es realizada en un solo sistema (fase móvil, columna cromatográfica, longitud de onda), diferenciándose sólo en la preparación de la muestra y el volumen de inyección por lo que no pueden ser cuantificadas en un solo cromatograma, debido a la gran diferencia de sus concentraciones en la tableta y sus propiedades de solubilidad. Previamente a la validación se evaluó la aptitud del sistema. Los resultados fueron conformes a las especificaciones para un método cromatográfico recomendadas por la USP 30, comprobando que el equipo, el sistema electrónico, las operaciones analíticas y las muestras a analizar constituyen un sistema integral que puede evaluarse como tal. Para la validación se evaluaron los parámetros de desempeño de la técnica como son: Selectividad, Exactitud, Precisión, Precisión Intermedia, Linealidad y Rango. Los resultados de estos parámetros se sometieron a pruebas estadísticas demostrando que la técnica analítica propuesta para la cuantificación de los principios activos es selectiva, exacta, precisa y lineal, así mismo la confiabilidad de la nueva técnica, garantizando de esta forma la calidad, eficacia e inocuidad del medicamento. / An analytical technique by high performance liquid chromatography (HPLC) was developed to quantify the active principles lysine clonixinate and pargeverine chlorhidrate in covered tablets, because the technique of analysis for this compound medicine is not in official books. The technique of analysis for both active principles is made in a single system (movable phase, chromatographic column, wavelength), being different itself only in the sample preparation and the volume of injection reason why cannot be quantified in a single chromatogram, due to the great difference of their concentrations in the tablet and its properties of solubility. Previously to the validation the aptitude of the system was evaluated. The results were in agreement to the specifications for a chromatographic method recommended by the USP 30, verifying that the equipment, the electronic system, the analytical operations and the samples to analyze constitute an integral system that can be evaluated like so. For the validation the parameters of performance of the technical were evaluated as they are: Selectivity, Accuracy, Precision, Intermediate Precision, Linearity and Range. The results of these parameters were also put to the test statistical demonstrating that the proposed analytical technical for the quantification of the active principles is selective, exact, precise and linear, to the trustworthiness of the new technical, guaranteeing of this form the quality, effectiveness and harmlessness of the medicine. / Tesis

Unidad de práctica para optar al título de Químico Farmacéutico / Los medicamentos esenciales son los productos farmacéuticos indispensables que cubren las necesidades sanitarias básicas de la población, y que son primordiales para el acceso a la salud. En Chile desde el año 1969, existe una lista con los medicamentos esenciales denominado Formulario Nacional de Medicamentos (FNM), el cual se actualizó por última vez en el año 2005. Este trabajo tiene como objetivo realizar un diagnóstico sobre la utilidad que tienen los medicamentos esenciales, tanto en Chile como en el resto del mundo; para generar una recomendación sobre el proceso, la actualización y la utilidad del FNM, incluyendo una propuesta de criterios para la inclusión de medicamentos en el Petitorio Mínimo exigido a las farmacias del sector privado. La Organización Mundial de la Salud (OMS) entrega cada dos años una lista modelo de medicamentos esenciales, los cuales son una guía para las Lista de Medicamentos Esenciales de cada nación, en las que existen criterios de selección, además de la formación de una comisión o comité multidisciplinario asesor que contribuye en la realización y actualización de dicha lista. En Chile la legislación indica cuales son los criterios de selección para la elaboración del FNM, y que son similares a los de la OMS. Sin embargo, a diferencia de la OMS, Chile no ha actualizado su FNM desde el año 2005, por lo que cuenta con productos que no están en el comercio o que están sin un registro sanitario vigente, dejando a la población sin el suministro o la disponibilidad de un producto esencial para su salud. Por lo tanto, para que el FNM cumpla con su objetivo, es indispensable que se revise y se actualice periódicamente, optimizando los recursos económicos en salud, mayor suministro y disponibilidad de medicamentos, entre otros; para aportar con un sistema de salud de calidad y el acceso a la salud de las personas. Este trabajo plantea una recomendación de la actualización del FNM, proponiendo la eliminación de algunos medicamentos basados en la recomendación de la OMS, arsenales fármaco-terapéuticos entre otros; también se recomienda dar cumplimiento a la exigencia de las monografías oficiales que se exigen y se anexan al Formulario, para poder entregar información completa del tratamiento los medicamentos y utilizarlo como una herramienta para el Uso Racional de medicamentos. Por otro lado, se recomienda al Ministerio de Salud que se incorporen de forma regular y periódica las actividades y tareas que demanda el Formulario, asignando los recursos humanos y materiales necesarios que den cumplimiento a la exigencia reglamentaria, en un trabajo coordinado entre los distintos actores del Ministerio de Salud. Finalmente, se recomienda establecer una estrecha red de colaboración entre los distintos actores interministeriales; todo esto servirá como herramienta para la optimización y gestión de compras, suministro y disponibilidad de medicamentos esenciales para contribuir a la salud de las personas

La presencia de medicamentos en los hogares se relacionada a un conjunto de factores. Se plantea como objetivo determinar los factores que de manera conjunta se relacionan con la tenencia de medicamentos y con el uso racional en los hogares. Se realizó un estudio observacional con diseño de campo trasversal, se aplicó 1092 encuestas, durante el periodo abril- julio 2009. Se realizó un muestro intencional por parroquias del área metropolitana del Municipio Trujillo, Estado Trujillo Venezuela. El instrumento se estructuró en las dimensiones: socio-económicas, condición de salud, sistema de salud. Para el análisis se aplicó técnicas estadísticas descriptivas, el chi cuadrado y log lineal, análisis de correspondencia mediante SPSS versión 10 y SPAD. Se encontró la asociación conjunta entre el número de medicamentos contabilizados en el hogar a un nivel de confianza de 95%(IC); con las variables: presencia de enfermos (agudos o crónicos): odds ration(OR) 2,5 p:0,000 (IC:1,9-3,33); ingreso familiar (en sueldos mínimo): OR:1,38 p:0,021, IC:1,05-1,82 y la compra de medicamentos(con o sin prescripción): OR:2,5 p:0,000 IC:1,53-4,04. Persiste en la población conductas irracionales como almacenamiento (86%), utilización de medicamentos con desconocimiento del uso (27,5%), vencidos (15%), sin prescripción (16%), automedicación (11%); utilización de tratamientos alternativos (25%).Es necesario diseñar políticas sanitarias de acceso a los medicamentos en la familia, considerando la presencia de enfermedad así como el ingreso familiar. Se debe fomentar en el médico el uso racional de medicamentos fortaleciendo el conocimiento sobre productos naturales, tratamientos caseros y actualizaciones farmacológicas, mediante campañas educativas. Palabras Clave: medicamentos, enfermedad, socioeconómico, uso, costo / The presence of drugs at home is related to a set of factors. The objective of this work is to determine the factors that jointly are related to the possession of drugs and their correct use at home. An descriptive study with a transverse field design was performed, applying 1092 surveys during the period April-July 2009. An intentional sampling was made in local parishes located in the metropolitan area of the Trujillo municipality, Trujillo State, Venezuela. The instrument was structured in the following dimensions: social-economical, health condition, health system. For the analysis, it was applied descriptive statistical techniques, the chi square and log-linear, correspondence analysis using SPSS 10 and SPAD. The overall association was found between the number of drugs counted at home at a confidence level of 95% (IC); with the variables: presence of sick people (chronic or acute): odds ration (OR) 2.5 p: 0.0001 (IC: 1.9-3.33); household income (minimum wage): OR: 1.38 p: 0.021, IC: 1.05-1.82 and the purchase of drugs (with or without prescription): OR: 2.5 p: 0.0001 IC: 1.53-4.04. Persists in the population irrational behaviours such as storage (86%), ignorance about the correct drug use (27.5%), expired drugs (15%), without prescription (16%), self-medication (11%), and use of alternative treatments (25%). It is necessary the design of health policies to have access to drugs in the family considering the presence of illness and family income. It must be encouraged the rational use of medical drugs by physicians to strengthen the knowledge about natural products, home treatments and pharmacological updates through educational campaigns. Key words: medical drugs, illness, socio-economic status, use, cost.

Se desarrolló y validó una nueva técnica de análisis por HPLC, que resultó ser efectiva, reproducible y confiable para la identificación y cuantificación de norfloxacino y fenazopiridina clorhidrato, asociados como principios activos en cápsulas orales. La técnica de análisis tomó en cuenta la preparación de muestra para que ambos activos puedan ser cuantificados en un solo sistema (fase móvil, columna cromatográfica, longitud de onda). Como paso previo a la validación de la técnica de análisis, se evaluó la adaptabilidad del sistema cromatográfico, asegurando de esta manera, el funcionamiento adecuado del sistema. Para el desarrollo de los parámetros de validación, se tomaron en cuenta los siguientes parámetros: Selectividad, Linealidad, Exactitud, Precisión y Robustez; resultados que fueron sometidos a evaluación estadística corroborando que la técnica analítica propuesta para la cuantificación de los principios activos es selectiva, lineal, exacta, precisa y robusta, así mismo la confiabilidad de la nueva técnica, garantizando de esta forma la calidad, eficacia e inocuidad del medicamento. Palabras Clave: Norfloxacino, fenazopiridina clorhidrato, cromatografía, validación, técnica analítica / It was developed and validated a new technique for HPLC analysis, which proved to be effective, reproducible and reliable for the identification and quantification of norfloxacin and phenazopyridine hydrochloride, associated as active partners in oral capsules. The technique of analysis took into account sample preparation for both assets can be quantified in a single system (mobile phase chromatographic column, wavelength). As step before to the validation of the technique of analysis, there was assessed the adaptability of the chromatographic system, assuring hereby, the suitable functioning of the system. For the development of validation parameters were taken into account the following parameters: selectivity, linearity, accuracy, precision and robustness; results that were submitted to statistical evaluation which confirmed that the proposed analytical technique for the quantification of active principles is selective, linear, accurate, precise and robust, likewise the reliability of the new technique, guaranteeing the quality, efficacy and safety of the drug. Key Words: Norfloxacin, phenazopyridine hydrochloride, high-resolution liquid chromatography, validation, analytical technique.