Desenvolvimento de técnica miniaturizada de extração em fase sólida empregando polímeros condutores para a determinação de meropenem em matrizes biológicas
Data
2022-12-06
Autores
Barbosa, João Antonio 
Orientadores
Calixto, Leandro Augusto
Tipo
Trabalho de conclusão de curso
Título da Revista
ISSN da Revista
Título de Volume
Resumo
A extração em fase sólida magnética (MSPE) é uma técnica miniaturizada derivada da extração em fase sólida (SPE), cuja diferença consiste no uso de sorventes magnetizados como polímeros condutores e polímeros magnéticos molecularmente não impressos, em inglês: magnetic non-imprinted polymers (MNIPs); podendo ser aplicada, por exemplo, na extração de fármacos em amostras biológicas, devido sua simplicidade na separação do analito-sorvente. Neste trabalho foi otimizada a MSPE, investigando o tipo e a quantidade de sorvente, o volume e pH da amostra, tipo e volume de solvente de eluição, e o efeito do sal, com o objetivo de extrair o meropenem de amostras constituídas com saliva artificial, visando a aplicação futura deste método de extração em amostras de saliva humana, utilizadas no monitoramento terapêutico do meropenem. A separação cromatográfica também foi otimizada considerando os seguintes parâmetros: volume de injeção, fluxo, comprimento de onda, coluna e a fase móvel. A MSPE foi executada nas amostras em triplicata, fortificadas com 200 μL de padrão meropem com concentração de 148 μg/mL. E o solvente de eluição, contendo o meropenem extraído, foi separado da fase sólida através da aplicação de um campo magnético (imã de neodímio), sendo ele levado posteriormente para análise no cromatógrafo líquido de alta eficiência com detecção ultravioleta. Ao final da otimização da MSPE, foram considerados adequados os seguintes parâmetros: 100 mg do MNIP com 4-vinilpiridina, adição de NaCl 10% na amostra com pH 7,0, e 500 μL de metanol como solvente de eluição. A recuperação obtida na condição otimizada foi de aproximadamente 3,7% do meropenem, uma quantidade considerada abaixo da faixa ideal, porém, o uso desta técnica de extração se mostrou promissora para aplicação no monitoramento terapêutico, visto que detém um menor tempo de execução. E além disso, este trabalho evidenciou que é possível uma melhoria na recuperação do meropenem por meio de testes com sorventes, por exemplo, mais seletivos.
Magnetic solid phase extraction (MSPE) is a miniaturized technique derived from solid phase extraction (SPE), whose difference consists in the use of magnetized sorbents such as conductive polymers and magnetic non imprinted polymers (MNIPs); it can be applied, for example, in the extraction of drugs in biological samples, due to its simplicity in the separation of the analyte-sorbent. In this work, MSPE was optimized, investigating the type and amount of sorbent, the volume and pH of the sample, type and volume of elution solvent, and the effect of salt, with the aim of extracting meropenem from samples constituted with artificial saliva, aiming at the future application of this extraction method in human saliva samples, used in the therapeutic monitoring of meropenem. The chromatographic separation was also optimized considering the following parameters: injection volume, flow, wavelength, column and mobile phase. MSPE was performed on samples in triplicate spiked with 200 μL of meropem standard at a concentration of 148 μg/mL. The elution solvent, containing the extracted meropenem, was separated from the solid phase through the application of a magnetic field (neodymium magnet), which was subsequently taken for analysis in a high-performance liquid chromatograph with ultraviolet detection. At the end of MSPE optimization, the following parameters were considered adequate: 100 mg of MNIP with 4-vinylpyridine, addition of 10% NaCl in the sample at pH 7.0, and 500 μL of methanol as elution solvent. The recovery obtained in the optimized condition was approximately 3.7% of meropenem, an amount considered below the ideal range, however, the use of this extraction technique proved to be promising for application in therapeutic monitoring, since it has a shorter execution time. Furthermore, this work showed that it is possible to improve the recovery of meropenem through tests with more selective sorbents, for example.
Magnetic solid phase extraction (MSPE) is a miniaturized technique derived from solid phase extraction (SPE), whose difference consists in the use of magnetized sorbents such as conductive polymers and magnetic non imprinted polymers (MNIPs); it can be applied, for example, in the extraction of drugs in biological samples, due to its simplicity in the separation of the analyte-sorbent. In this work, MSPE was optimized, investigating the type and amount of sorbent, the volume and pH of the sample, type and volume of elution solvent, and the effect of salt, with the aim of extracting meropenem from samples constituted with artificial saliva, aiming at the future application of this extraction method in human saliva samples, used in the therapeutic monitoring of meropenem. The chromatographic separation was also optimized considering the following parameters: injection volume, flow, wavelength, column and mobile phase. MSPE was performed on samples in triplicate spiked with 200 μL of meropem standard at a concentration of 148 μg/mL. The elution solvent, containing the extracted meropenem, was separated from the solid phase through the application of a magnetic field (neodymium magnet), which was subsequently taken for analysis in a high-performance liquid chromatograph with ultraviolet detection. At the end of MSPE optimization, the following parameters were considered adequate: 100 mg of MNIP with 4-vinylpyridine, addition of 10% NaCl in the sample at pH 7.0, and 500 μL of methanol as elution solvent. The recovery obtained in the optimized condition was approximately 3.7% of meropenem, an amount considered below the ideal range, however, the use of this extraction technique proved to be promising for application in therapeutic monitoring, since it has a shorter execution time. Furthermore, this work showed that it is possible to improve the recovery of meropenem through tests with more selective sorbents, for example.