Analytical high performance liquid chromatograph

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Study of metabolic pathways requires the possibility to detect, identify, separate, and quantify various metabolites in complex mixtures. Mass spectrometry in combination with liquid chromatography is a powerful tool for resolving the composition of those complex mixtures. While our working group has an access to the instruments aimed for molecular structure elucidation at the Institute for Molecular Microbiology and Biotechnology, the main challenges for us are (i) reliable detection and quantification of polar metabolites (e.g., carbonic acids, phosphosugars), (ii) quantification of CoA-esters in the mixtures with other CoA-esters with similar chromatographic behaviour, (iii) purification of various, commercially unavailable CoA-esters for our research, and (iv) establishment of isotopologue profiling in our laboratory. Low purity of the samples that we typically analyse creates additional difficulties for us. Therefore, we need a robust, easy-to-use single quadrupole mass spectrometer connected to the liquid chromatography system optimized for the separation of polar compounds, prone to loss due unspecific molecular interactions. In terms of detection, the system needs to be equipped with photodiode array and fluorescence detectors. The automatic programmable fraction collector will significantly extend our capabilities of purification. This system is attractive for other working groups in our institute and beyond, and it will strengthen our collaboration network and lead to more interdisciplinary projects.