Desenvolvimento e validação de técnica bioanalítica em LC-MS/MS para detecção e quantificação de protótipos antineoplásticos

• Ano de defesa: 2014
• Autor(a) principal: Gomes, Sandro Antônio
• Orientador(a): Cunha, Luiz Carlos da
• Banca de defesa: Cunha, Luiz Carlos da , Efting, Cristiane, Salazar, Vânia Cristina Rodriguez, Teixeira, Leonardo de Souza
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Goiás
• Programa de Pós-Graduação: Programa de Pós-graduação em Ciências da Saúde (FM)
• Departamento: Faculdade de Medicina - FM (RG)
• País: Brasil
• Palavras-chave em Português: LC-MS/MS; LQFM018; LQFM030; Nutlins; Inibidores de MDM2-p53; Antitumorais
• Palavras-chave em Inglês: LC-MS/MS; LQFM018; LQFM030; Nutlins; Inhibitors of MDM2-p53; Antitumors
• Área do conhecimento CNPq: CIENCIAS DA SAUDE
• Link de acesso: http://repositorio.bc.ufg.br/tede/handle/tede/6521

A rapid, sensitive and selective liquid chromatography tandem mass spectrometry (LC/MS-MS) bioanalytical method was developed and validated to quantify LQFM018 and LQFM030 prototypes obtained by molecular simplification nutlins (inhibitors of MDM2-p53 interaction) in mouse, rat and human plasma. The prototypes and the internal standard domperidone were extracted from the plasma samples by liquid-liquid extraction using methyl tert-butyl ether. The chromatographic analysis was performed using ACE ® C18 analytical column (5µm 100 x 4.6 mm) for a total time of 4 minutes. In MRM method transitions (m/z) were used: 349.138/191.10, 319.162/191.20 and 426.032/175.20 Daltons to LQFM018; LQFM030 and internal standard, respectively. The mobile phase consists of methanol-2 mM ammonium acetate containing 0.025% formic acid. The calibration curve was linear over the concentration range examined 10- 15000 ng/mL with r> 0.99 and the limit of quantitation of 10 ng/mL for both prototypes. For LQFM018, intra-assay precision was 0.8 to 7.3% and accuracy from 96.8 to 105.8%. And inter-assay to the precision was 2.3 and 6.6% and accuracy was 99.3 to 104.3%. The average recovery was 65.0% and the matrix effect between -11.2 to 2.1%. To LQFM030, intra-assay precision was 0.6 to 5.5% and accuracy from 95.5 to 111.3%. Inter-assay precision was 1.8 to 6.7%, and accuracy was 99.0 to 107.0%. The average recovery was 74.1% and the matrix effect from -7.9 to 1.5%. Recovery for internal standard was 61.3%. The prototypes were considered stable in the biological matrix and the solution proposed tests. Thus, the analytical method was developed and validated is capable to quantify traces of concentration in plasma.