Desenvolvimento de técnicas miniaturizadas de extração em fase líquida empregando solventes eutéticos hidrofóbicos para determinação de meropenem em em saliva artificial e futura aplicação em matriz biológica
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Data
2023-11-29
Autores
Almeida, Maria Karilly Lima de [UNIFESP]
Orientadores
Calixto, Leandro Augusto [UNIFESP]
Tipo
Trabalho de conclusão de curso de graduação
Título da Revista
ISSN da Revista
Título de Volume
Resumo
O meropenem é um antibiótico de amplo espectro utilizado no tratamento de infecções causadas por bactérias gram-positivas e gram-negativas. Por ser um medicamento tempo dependente, sua eficácia está relacionada com o tempo de duração que a concentração sérica permanece acima da concentração inibitória mínima (CIM). Deste modo, desfechos clínicos insatisfatórios geralmente ocorrem pela administração insuficiente do fármaco, levando a níveis sérios que não atingem a CIM. O monitoramento terapêutico de fármacos (TDM) em matriz biológica surge nesse contexto como uma estratégia para minimizar a toxicidade e maximizar a eficácia do medicamento. Este trabalho teve por objetivo desenvolver uma metodologia analítica seguindo os princípios da química analítica verde para determinação desse antibiótico, empregando uma metodologia analítica desenvolvida por HPLC-UV em amostras de saliva artificial, visando futura aplicação em saliva de procedência biológica. A técnica de extração utilizada foi a microextração líquido-líquido dispersiva (DLLME) utilizando solvente eutético hidrofóbico (HDES, hydrophobic deeep eutectic solvents) como solvente extrator e o dietilcarbonato (carbonato orgânico verde) como solvente dispersor. A otimização do método foi realizada utilizando amostras com 800 µL de saliva e 200 µL de solução estoque de meropenem na concentração de 200 µg/mL. As condições de análise definidas foram: comprimento de onda de 298 nm, fase móvel MeOH:Tampão fosfato 0,1 M pH 3,14, na vazão de 1 mL/min, coluna Kromasil C18 5μm, 150 x 4,6 mm e eluição por gradiente. O meropenem eluiu em 12,7 minutos e sem a coeluição com potenciais interferentes. A triagem foi feita utilizando o software Minitab para uma abordagem multivariável empregando 8 parâmetros diferentes, cada um com 2 níveis. Observou-se que os fatores que mais interferiram na extração do meropenem foram pH, efeito do sal, volume do extrator e volume do dispersor, que seguiram para a etapa de otimização por Delineamento Composto Central (CCD), avaliando-se 5 níveis. A condição que obteve maior recuperação (7%) foi a com pH 2, sem adição de sal e extração realizada com 150 µL de solvente dispersor e 100 µL de solvente extrator. A extração do meropenem utilizando HDES se mostrou uma técnica simples e versátil para determinação do fármaco em saliva artificial, além de uma alternativa verde em comparação aos compostos orgânicos geralmente empregados.
Meropenem is a broad-spectrum antibiotic used to treat infections caused by gram-positive and gram-negative bacteria. As it is a time-dependent medication, its effectiveness is related to the length of time that the serum concentration remains above the minimum inhibitory concentration (MIC). Therefore, unsatisfactory clinical outcomes generally occur due to insufficient administration of the drug, leading to serious levels that do not reach the MIC. Therapeutic drug monitoring (TDM) in biological matrix emerges in this context as a strategy to minimize toxicity and maximize drug efficacy. This work aimed to develop an analytical methodology following the principles of green analytical chemistry to determine this antibiotic using an analytical methodology developed by HPLC-UV in artificial saliva samples, aiming for future application in saliva of biological origin. The extraction technique used was dispersive liquid-liquid microextraction (DLLME), using hydrophobic eutectic solvent (HDES, hydrophobic deep eutectic solvents) as the extracting solvent and diethylcarbonate (green organic carbonate) as the dispersing solvent. The optimization of the method was carried out using samples with 800 μL of saliva and 200 μL of meropenem stock solution at a concentration of 200 μg/mL. The defined analysis conditions were: wavelength of 298 nm, mobile phase MeOH:0.1M phosphate buffer pH 3.14, flow rate of 1 mL/min. Kromasil C18 5μm column, 150 x 4.6 mm and gradient elution. Meropenem eluted in 12.7 minutes and without co-elution with potential interference. Screening was done using Minitab software for a multivariable approach employing 8 different parameters, each with 2 levels. It was observed that the factors that most interfered with the extraction of meropenem were pH, salt effect, extractor volume and disperser volume, which went to the optimization stage by Central Composite Design (CCD), evaluating 5 levels. The condition that obtained the highest recovery (7%) was pH 2, without addition of salt and extraction carried out with 150 μL of dispersing solvent and 100 μL of extracting solvent. The extraction of meropenem using HDES proved to be a simple and versatile technique for determining the drug in artificial saliva, as well as a green alternative compared to the organic compounds generally used.
Meropenem is a broad-spectrum antibiotic used to treat infections caused by gram-positive and gram-negative bacteria. As it is a time-dependent medication, its effectiveness is related to the length of time that the serum concentration remains above the minimum inhibitory concentration (MIC). Therefore, unsatisfactory clinical outcomes generally occur due to insufficient administration of the drug, leading to serious levels that do not reach the MIC. Therapeutic drug monitoring (TDM) in biological matrix emerges in this context as a strategy to minimize toxicity and maximize drug efficacy. This work aimed to develop an analytical methodology following the principles of green analytical chemistry to determine this antibiotic using an analytical methodology developed by HPLC-UV in artificial saliva samples, aiming for future application in saliva of biological origin. The extraction technique used was dispersive liquid-liquid microextraction (DLLME), using hydrophobic eutectic solvent (HDES, hydrophobic deep eutectic solvents) as the extracting solvent and diethylcarbonate (green organic carbonate) as the dispersing solvent. The optimization of the method was carried out using samples with 800 μL of saliva and 200 μL of meropenem stock solution at a concentration of 200 μg/mL. The defined analysis conditions were: wavelength of 298 nm, mobile phase MeOH:0.1M phosphate buffer pH 3.14, flow rate of 1 mL/min. Kromasil C18 5μm column, 150 x 4.6 mm and gradient elution. Meropenem eluted in 12.7 minutes and without co-elution with potential interference. Screening was done using Minitab software for a multivariable approach employing 8 different parameters, each with 2 levels. It was observed that the factors that most interfered with the extraction of meropenem were pH, salt effect, extractor volume and disperser volume, which went to the optimization stage by Central Composite Design (CCD), evaluating 5 levels. The condition that obtained the highest recovery (7%) was pH 2, without addition of salt and extraction carried out with 150 μL of dispersing solvent and 100 μL of extracting solvent. The extraction of meropenem using HDES proved to be a simple and versatile technique for determining the drug in artificial saliva, as well as a green alternative compared to the organic compounds generally used.