RESUMO
A bioanalytical method for the quantitative determination of budesonide and fluticasone in human sputum was developed. Sputolysin(®) Reagent was added to the sputum samples. After incubation (37°C; 60-70 min under shaking) and automated solid phase extraction the extracts were analysed using LC-MS/MS. Budesonide and fluticasone showed good linearity (r>0.99) over the range 0.1-100 nM in the first and second validation batch, and over the range 0.25-10,000 nM in the third and fourth validation batch. The lower limit of quantification (LLOQ) achieved was 5 nM for budesonide and fluticasone in 100 µL human sputum. Intra-run and inter-run RSD for four quality control levels (5-100 nM) were within 6.9% (budesonide) and 8.0% (fluticasone). The accuracy ranged from -11.4% to -1.6% (budesonide), and from -11.8% to 0.4% (fluticasone). The validated method was applied to clinical sputum samples from COPD patients.
Assuntos
Androstadienos/análise , Budesonida/análise , Cromatografia Líquida/métodos , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/metabolismo , Escarro/química , Espectrometria de Massas em Tandem/métodos , Androstadienos/farmacocinética , Androstadienos/uso terapêutico , Anti-Inflamatórios/análise , Anti-Inflamatórios/farmacocinética , Anti-Inflamatórios/uso terapêutico , Budesonida/farmacocinética , Budesonida/uso terapêutico , Estabilidade de Medicamentos , Fluticasona , Humanos , Modelos Lineares , Reprodutibilidade dos Testes , Extração em Fase Sólida , Escarro/metabolismo , TemperaturaRESUMO
An LC-MS/MS method has been developed for the quantitative determination of a protein drug (Tenecteplase; M(W) 58,777 Da) in rat plasma. The protein was digested with trypsin without prior clean-up of the plasma sample, without the use of a label nor internal standard. A limited validation was performed to assess the linearity, the sensitivity and the specificity of the method. In addition, the developed method was applied to the quantitative analysis of Tenecteplase in rat plasma samples originating from a single-dose study in rats.