Liquid chromatography coupled with mass spectrometry (LC-MS) is one of the most powerful analytical techniques, enabling efficient separation, identification, and quantification of a wide range of chemical compounds in various samples.
As part of the master's thesis, we developed and optimized methods based on liquid chromatography coupled with mass spectrometry (LC-MS) for selected compounds. The goal was to develop analytical methods that are time-efficient, accurate, selective, and robust.
We first developed the methods using compound standards. We optimized chromatographic conditions and mass spectrometric parameters for all selected compounds. For method optimization and analysis, we used a Thermo Scientific Vanquish Core HPLC system coupled with a TSQ Fortis Plus triple quadrupole mass spectrometer.
After optimization, we analyzed samples consisting of cell lysates after incubation with the selected compounds. In cases where the method did not allow detection of sufficiently low concentrations, we did not construct calibration curves or evaluate linearity, accuracy, precision, and the limit of detection.
For samples with compound UL-EID-11, no sample preparation was needed, while for compounds 6, 8, 12, and 28, proteins were removed by precipitation. For compound UL-EID-11 and compound 6, we developed a sufficiently sensitive method that enabled quantification of analyte concentrations in the samples. However, for compounds 8, 12, and 28, the analyte concentrations in the samples were not high enough to be detected by the optimized analytical method.
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