Focus on the ability of synthesize libraries of structurally different compounds was given through discovery and need for quick and effective search of new active substances. For the discovery of new active substances, is most commonly used the High Throughput Screening (HTS) method, from which other related concepts of synthetic chemistry have been developed based on the design of the structure and properties of molecules. In the last decade, the number of new detected compounds has increased significantly, but nevertheless, we continue to use the classic approaches. In addition to HTS libraries we use the so-called in-house libraries.
The aim of this thesis was to develop and optimize HPLC method coupled with a mass detector to identify and quantify the compounds from the library of Faculty of pharmacy.
The main focus was on the development of the standard HPLC method, and on the large number of prepared compounds, for which we checked the effectiveness of our method. In particular, we investigated the impact of various chromatographic conditions, such as the composition of the mobile phase, the choice of an appropriate chromatographic column, and the choice of the appropriate gradient, which would allow the universal use. Using the HPLC / MS method, we were able to identify 80% of the investigated compounds. Method is therefore suitable for qualitative and quantitative evaluation. A key part of the research work was to manually verify the results obtained, since modern methods do not allow automated evaluation of data. The data were statistically evaluated and identified for the main compound and also its disintegrating products. By using the mass spectrometry, we could also predict what happened to the investigated compound and which chemical reaction has occurred before or after analyzing the compounds. The chromatographic conditions were not suitable for the remaining 20% of the investigated compounds, thus should be considered individually.
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