Abstract
The study optimized the analytical method for determining the active components of targeted drugs for the treatment of low-grade gliomas dabrafenib and trametinib, in a model organoid system. Third generation organoids were incubated for 24 hours in the presence of 100 μg/ml dabrafenib and trametinib (substance, SIGMA-Aldrich, Germany) (3 cultures in each series). The method for determining dabrafenib and trametinib in organoid tissue included sequential washing of tumor-like structures from the incubation medium, homogenization and centrifugation, followed by determination of the concentration of active substances in the supernatant by HPLC on a Shimadzu LCMS-8030 mass spectrometer (Japan) using a Phenomenex Luna® C18 column (2) 250 * 4.6 mm, with an internal diameter of 5 μm and a sorbent pore diameter of 100 Å. The release time (RT) of dabrafenib in these conditions was 7.405 minutes, the RT of trametinib was 8.356 minutes, the selectivity factor (α) was 1.19, which satisfied the requirements (α > 1). The chromatographic system resolution (Rs) was 2.13, indicating complete separation (Rs > 1.5). Thus, the developed method may be used to determine molecularly targeted drugs - dabrafenib and trametinib in model tumor systems.
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