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Abstract(s)
O relatório de estágio aqui apresentado está inserido na unidade curricular “Estágio” do Mestrado Integrado em Ciências Farmacêuticas e encontra-se divido em três capítulos. O primeiro capítulo é referente ao estágio curricular em Farmácia Hospitalar, realizado nos Serviços Farmacêuticos do Centro Hospitalar Universitário Cova da Beira. Já o segundo capítulo aborda a minha experiência durante o estágio em Farmácia Comunitária e decorreu na Farmácia Moderna de Castro Daire. Em ambos são abordadas todas as atividades desenvolvidas, com destaque especial para o papel do farmacêutico, que desempenha cada vez mais um papel fundamental ao nível dos cuidados de saúde e promoção da saúde pública. Por último, no terceiro capítulo é descrito todo o trabalho de investigação laboratorial desenvolvido no Centro de Investigação em Ciências da Saúde da Universidade da Beira Interior.
O presente trabalho descreve o desenvolvimento e validação de um novo método analítico para a determinação de metadona e do seu principal metabolito EDDP em amostras de fluido oral, com recurso a Dried Saliva Spots (DSS) e cromatografia gasosa acoplada à espectrometria de massa em tandem (GC-MS/MS).
O procedimento de extração da amostra consistiu na aplicação de uma amostra de fluido oral (50 µL) num cartão de papel de filtro, seguindo-se da adição de isopropanol e agitação. Posteriormente, para obtenção do extrato, foi necessário ocorrer centrifugação e concentração.
Decorrido este pré-tratamento, o extrato foi injetado na coluna cromatográfica, sendo que neste trabalho, para favorecer a separação dos analitos, foi utilizada uma coluna de fenilmetilsiloxano a 5%.
Uma vez que era desejado que o sinal obtido apresentasse o menor ruído possível, todas as condições cromatográficas e parâmetros de espectrometria de massa foram otimizados. Relativamente à validação do método foram utilizados critérios aceites internacionalmente para validação de métodos bioanaliticos, sendo que os parâmetros estudados incluíram o estudo da seletividade, linearidade, limites de deteção (LOD) e quantificação (LLOQ), precisão e exatidão, estabilidade, fatores de diluição e recuperação. O procedimento foi linear para o intervalo de concentrações de 10 a 250 ng/mL para ambos os compostos, com coeficientes de determinação (R2) superiores a 0,99. A precisão intra- e interdia foram inferiores a 15% para todos os analitos a todas as concentrações estudadas, enquanto a exatidão permaneceu dentro de um intervalo de ± 15%. As recuperações variaram de 45 a 74%. Utilizando apenas 50 µL de amostra foi possível obter LODs baixos (5 ng/mL).
Todos os parâmetros estudados obedeceram aos critérios definidos e o método permitiu a determinação bem-sucedida da metadona e do seu principal metabolito em amostras de fluído oral de indivíduos em terapia de substituição/manutenção de opiáceos.
The internship report presented here is included in the curricular unit "Internship" of the Integrated Master in Pharmaceutical Sciences and is divided into three chapters. The first chapter refers to the curricular internship in Hospital Pharmacy, held at the Pharmaceutical Services of the Centro Hospitalar Universitário Cova da Beira. The second chapter addresses the topic of my experience during the internship in Community Pharmacy, which took place at Farmácia Moderna Castro Daire. Both focus on all the activities developed, with particular emphasis on the role of the pharmacist, who plays an increasingly important role in health care and public health promotion. Finally, the third chapter describes all laboratory research work developed at Centro de Investigação em Ciências da Saúde da Universidade da Beira Interior. The present work describes the development and validation of new analytical method for the determination of methadone and its main metabolite EDDP in oral fluid samples, using the Dried Saliva Spots (DSS) sampling approach, by Gas Chromatography in tandem mass spectrometry (GC-MS/MS). The sample extraction procedure was carried out by spotting the oral fluid sample (50 µL) onto a filter paper card, and immersion in isopropyl alcohol and shaking, followed by centrifugation and extract concentration. After this pre-treatment, the solution obtained is ready for injection in the chromatographic column, and in this work, separation was achieved using a 5% phenylmethylsiloxane column. All chromatographic conditions and mass spectrometric parameters were optimized to enhance the signal. Regarding the validation of the method, internationally accepted criteria were considered, and the studied parameters included selectivity, linearity, limits of detection (LOD) and quantification (LLOQ), precision and accuracy, stability, dilution integrity and recovery. The procedure was linear for concentrations ranging from 10 to 250 ng/mL for both compounds, with determination coefficients (R2) higher than 0.99. Intra- and inter-day precision was lower than 15% for all analytes at the studied concentrations, while accuracy remained within a ± 15% interval from the nominal concentrations. Recoveries ranged from 45 to 74%. Low LODs were achieved for both compounds (5 ng/mL), using only 50 µL as sample volume. All studied parameters complied with the defined criteria and the sampling approach enabled the successful determination of methadone and its main metabolite in low volume oral fluid samples of patients undergoing opiate substitution/maintenance treatment.
The internship report presented here is included in the curricular unit "Internship" of the Integrated Master in Pharmaceutical Sciences and is divided into three chapters. The first chapter refers to the curricular internship in Hospital Pharmacy, held at the Pharmaceutical Services of the Centro Hospitalar Universitário Cova da Beira. The second chapter addresses the topic of my experience during the internship in Community Pharmacy, which took place at Farmácia Moderna Castro Daire. Both focus on all the activities developed, with particular emphasis on the role of the pharmacist, who plays an increasingly important role in health care and public health promotion. Finally, the third chapter describes all laboratory research work developed at Centro de Investigação em Ciências da Saúde da Universidade da Beira Interior. The present work describes the development and validation of new analytical method for the determination of methadone and its main metabolite EDDP in oral fluid samples, using the Dried Saliva Spots (DSS) sampling approach, by Gas Chromatography in tandem mass spectrometry (GC-MS/MS). The sample extraction procedure was carried out by spotting the oral fluid sample (50 µL) onto a filter paper card, and immersion in isopropyl alcohol and shaking, followed by centrifugation and extract concentration. After this pre-treatment, the solution obtained is ready for injection in the chromatographic column, and in this work, separation was achieved using a 5% phenylmethylsiloxane column. All chromatographic conditions and mass spectrometric parameters were optimized to enhance the signal. Regarding the validation of the method, internationally accepted criteria were considered, and the studied parameters included selectivity, linearity, limits of detection (LOD) and quantification (LLOQ), precision and accuracy, stability, dilution integrity and recovery. The procedure was linear for concentrations ranging from 10 to 250 ng/mL for both compounds, with determination coefficients (R2) higher than 0.99. Intra- and inter-day precision was lower than 15% for all analytes at the studied concentrations, while accuracy remained within a ± 15% interval from the nominal concentrations. Recoveries ranged from 45 to 74%. Low LODs were achieved for both compounds (5 ng/mL), using only 50 µL as sample volume. All studied parameters complied with the defined criteria and the sampling approach enabled the successful determination of methadone and its main metabolite in low volume oral fluid samples of patients undergoing opiate substitution/maintenance treatment.
Description
Keywords
Dried Saliva Spots Eddp Farmácia Comunitária Farmácia Hospitalar Fluído Oral Gc/Ms/Ms. Metadona