RESUMEN
Dabrafenib is an inhibitor of BRAF V600E used for treating metastatic melanoma but a majority of patients experience adverse effects. Methods to measure the levels of dabrafenib and major metabolites during treatment are needed to allow development of individualized dosing strategies to reduce the burden of such adverse events. In this study, an LC-MS/MS method capable of measuring dabrafenib quantitatively and six metabolites semi-quantitatively is presented. The method is fully validated with regard to dabrafenib in human plasma in the range 5-5000 ng/mL. The analytes were separated on a C18 column after protein precipitation and detected in positive electrospray ionization mode using a Xevo TQ triple quadrupole mass spectrometer. As no commercial reference standards are available, the calibration curve of dabrafenib was used for semi-quantification of dabrafenib metabolites. Compared to earlier methods the presented method represents a simpler and more cost-effective approach suitable for clinical studies. Graphical abstract Combined multi reaction monitoring transitions of dabrafenib and metabolites in a typical case sample.
Asunto(s)
Antineoplásicos/sangre , Antineoplásicos/metabolismo , Imidazoles/sangre , Imidazoles/metabolismo , Oximas/sangre , Oximas/metabolismo , Espectrometría de Masas en Tándem/métodos , Cromatografía Líquida de Alta Presión/economía , Cromatografía Líquida de Alta Presión/métodos , Monitoreo de Drogas/economía , Monitoreo de Drogas/métodos , Humanos , Límite de Detección , Microsomas Hepáticos/metabolismo , Reproducibilidad de los Resultados , Espectrometría de Masas en Tándem/economíaRESUMEN
Non-linear microscopy has been used to characterize bovine smooth muscle cells and their proliferation, migration, and differentiation in hydrogel cellulose scaffolds, toward the development of fully functional blood vessel implants. The extracellular matrix (ECM) composed of cellulose and endogenous collagen fibers was imaged using Second Harmonic Generation (SHG) microscopy and the cell morphology by Coherent Anti-Stokes Raman Scattering (CARS) microscopy. Images prove that cells adhere on the cellulose scaffold without additional surface modification and that both contractile and proliferating phenotypes are developed. This work shows that non-linear microscopy contributes with unique insights in cell interactions with (artificial) ECM components and has the potential to become an established characterization method in tissue engineering.
