Tato práce prezentuje vývoj a validaci komplexní analytické metody (UHPLC-MS/MS) umožňující kvantifikaci vybrané skupiny steroidů s neuroaktivními účinky v lidském krevním séru. Kromě toho bylo prováděno profilování vybraných metabolitů tryptofanu v séru a mozkomíšním moku různých neuropatologických skupin s cílem zmapovat jejich metabolické změny. Na závěr byly také studovány vybrané kandidátní proteinové biomarkery.One of the current hot topics is the increasing proportion of neurodegenerative diseases. The key challenge is to design reliable diagnostic tools capable of detecting these diseases before they progress to an advanced stage. Neuroactive steroids, which have the potential to modulate the development of the nervous system, represent a highly promising avenue of research. Therefore, the availability of dependable methods for monitoring changes in the concentration of these substances is of paramount importance. This study presents the development and validation of a comprehensive method based on ultra-high performance liquid chromatography-tandem mass spectrometry for the quantification of a selected group of steroids with neuroactive effects in human serum. Additionally, this research involved profiling a range of tryptophan-related compounds in the serum and cerebrospinal fluid of various neuropathological cohorts to map their metabolic alterations. Furthermore, selected candidate protein biomarkers were studied.