High-resolution mass spectrometer with Liquid Chromatography (LC) and Capillary Electrophoresis (CE)

通过例如液相色谱法（LC-MS）分离化合物的质谱法已经成为大规模代谢组学测量的关键方法。与直接注入质谱法相比，LC-MS提供了几个优点，因为来自复杂样品基质的化合物被分离成不同的峰，这减少了离子抑制并提高了灵敏度。该提案旨在采购一种新的串联质谱仪，用于采用靶向和非靶向策略对小分子进行LC-MS和毛细管电泳-质谱（CE-MS）测量。新仪器将用于识别和定量信号分子（例如，磷酸肌醇和肌醇-磷酸），次级植物代谢物（例如，类黄酮）和其他分子，重点是植物和细菌。我们将使用LC-MS建立一个管道，用于鉴定在两个生物系统之间差异积累的代谢物（非靶向代谢组学），例如通过比较野生型和突变体，或暴露于对照或应激条件的生物体。仪器将切换至CE-MS模式，以测量肌醇磷酸盐和其他极性阴离子代谢物（NADPH、ATP等）。波恩大学校园目前没有CE-MS系统。该质谱仪将位于波恩大学植物分子生理学和生物技术研究所（IMBIO）现有的脂质组学/代谢组学平台中。该平台已经运行了两台LC-MS仪器，用于高容量的直接输注/枪式测量甘油脂，以及低丰度代谢物的LC-MS分析。新的质谱仪将用于高质量准确度和灵敏度的测量，这是识别和定量复杂提取物中大量代谢物所需的。该工具对于已获得资助或正在评估的几项（联合）赠款申请至关重要。质谱仪将服务于校园内的团体，但它也将开放与国家和国际研究团体的合作。

Mass spectrometry with separation of compounds by e.g. liquid chromatography (LC-MS) has emerged as a key method for large scale metabolomic measurements. In contrast to direct infusion mass spectrometry, LC-MS offers several advantages because compounds from complex sample matrices are separated into different peaks, which reduces ion suppression and results in improved sensitivity. This proposal aims at the procurement of a new tandem mass spectrometer for LC-MS and capillary electrophoresis-mass spectrometry (CE-MS) measurements of small molecules, both employing targeted and non-targeted strategies. The new instrument will be used for the identification and quantification of signaling molecules (e.g., phosphoinositides and inositol-phosphates), secondary plant metabolites (e.g., flavonoids) and other molecules, with a focus on plants and bacteria. We will use the LC-MS to establish a pipeline for the identification of metabolites that differentially accumulate between two biological systems (non-targeted metabolomics), e.g. by comparing wild types and mutants, or organisms exposed to control or stress conditions. The instrument will be switched to CE-MS mode for the measurement of inositol-phosphates and other polar, anionic metabolites (NADPH, ATP, etc.). The CE-MS system is currently not available on campus at the University of Bonn. The mass spectrometer will be located and integrated into the existing Lipidomics/Metabolomics Platform at the Institute of Molecular Physiology and Biotechnology of Plants (IMBIO) of the University of Bonn. This platform already operates two LC-MS instruments, which are used at high capacity for direct infusion/shotgun measurements of glycerolipids, and LC-MS analysis of low abundant metabolites. The new mass spectrometer will be used for measurements with high mass accuracy and sensitivity, which is required for the identification and quantification of large numbers of metabolites in complex extracts. The instrument will be essential for several (joint) grant applications that have been funded or are under evaluation. The mass spectrometer will serve groups on campus, but it will also be open to collaborations with national and international research groups.