Analytical high performance liquid chromatograph

代谢途径的研究需要能够检测、识别、分离和定量复杂混合物中的各种代谢物。质谱与液相色谱相结合是解析这些复杂混合物组成的有力工具。虽然我们的工作组可以使用分子微生物学和生物技术研究所用于分子结构解析的仪器，但我们面临的主要挑战是（i）极性代谢物的可靠检测和定量（例如，碳酸，磷酸糖），（ii）与具有类似色谱行为的其他CoA-酯的混合物中的CoA-酯的定量，（iii）用于我们研究的各种市售CoA-酯的纯化，和（iv）在我们实验室中建立同位素体谱。我们通常分析的低纯度样品给我们带来了额外的困难。因此，我们需要一个强大的，易于使用的单四极杆质谱仪连接到液相色谱系统优化的极性化合物的分离，容易损失由于非特异性分子相互作用。在检测方面，系统需要配备光电二极管阵列和荧光检测器。自动可编程馏分收集器将大大扩展我们的纯化能力。该系统对我们研究所内外的其他工作组具有吸引力，它将加强我们的合作网络，并导致更多的跨学科项目。

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.