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

• Autor(a) principal: Gomes, Sandro Antônio
• Data de Publicação: 2014
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional da UFG
• dARK ID: ark:/38995/001300000bqcp
• Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/6521
• topic: LC-MS/MS; LQFM018; LQFM030; Nutlins; Inibidores de MDM2-p53; Antitumorais; LC-MS/MS; LQFM018; LQFM030; Nutlins; Inhibitors of MDM2-p53; Antitumors; CIENCIAS DA SAUDE

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.